Addgene 1137=Hsp70Bb-Cas9[dsRed] Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: 1137=Hsp70Bb-Cas9[dsRed] Sequencing Result


Map View

M13 Forward (12,743 .. 12,760) M13/pUC Forward (12,729 .. 12,751) NdeI (12,549) pRS-marker (12,516 .. 12,535) pGEX 3' (12,394 .. 12,416) pBRforEco (12,338 .. 12,356) Amp-R (11,928 .. 11,947) pBR322ori-F (10,626 .. 10,645) L4440 (10,375 .. 10,392) CAP binding site lac promoter M13/pUC Reverse (10,157 .. 10,179) lac operator M13 Reverse (10,144 .. 10,160) M13 rev EcoRI (10,130) KpnI (10,122) Acc65I (10,118) BamHI (10,109) piggyBac left (5') inverted repeat EBV-rev (9231 .. 9250) SV40pA-R (9177 .. 9196) DsRed1-C (8941 .. 8964) StuI (8781) NcoI (8767) mCherry-F (8747 .. 8766) SbfI (8689) DsRed1-N (8529 .. 8549) FspAI (8397) RsrII (8053) AscI (7798) PacI (7116) EGFP-C (7045 .. 7066) M13 fwd AgeI (1156) BspDI - ClaI (1269) AvrII (1375) SacII (1411) PflFI - Tth111I (1596) PaeR7I - XhoI (1664) NruI (1808) pCasper-hs (2005 .. 2026) PmeI (2061) BglII (2436) PasI (3955) PmlI (4975) EXFP-R (6698 .. 6717) 1137=Hsp70Bb-Cas9[dsRed] 13,159 bp

Sequence View

5ʹ 3ʹ 60 DTCSRTNVCLAFYLLTLFYI c g t c t g t a c a c g a a c g c g t a t t a a c g c a g a g t g c a a a g t a t a a g a g g g t t a a a a a a t a t a g c a g a c a t g t g c t t g c g c a t a a t t g c g t c t c a c g t t t c a t a t t c t c c c a a t t t t t t a t a t

120 KRVM t t t t a c g c a c c a t a t a c g c a t c g g g t t g a t a t c g t t a a t a t g g a t c a a t t t g a a c a g t t g a a a a t g c g t g g t a t a t g c g t a g c c c a a c t a t a g c a a t t a t a c c t a g t t a a a c t t g t c a a c

180 a t t a a c g t g t c t c t g c t c a a g t c t t t g a t c a a a a c g c a a a t c g a c g a a a a t g t g t c g g a c t a a t t g c a c a g a g a c g a g t t c a g a a a c t a g t t t t g c g t t t a g c t g c t t t t a c a c a g c c t g

240 a a t a t c a a g t c g a t g a g c g a a a a a c t a a a a a g g c t a g a a t a c g a c a a t c t c a c a g a c a g c t t a t a g t t c a g c t a c t c g c t t t t t g a t t t t t c c g a t c t t a t g c t g t t a g a g t g t c t g t c g

300 g t t g a g a t a t a c g g t a t t c a c g a c a g c a g g c t g a a t a a t a a a a a a a t t a g a a a c t a t t a t c a a c t c t a t a t g c c a t a a g t g c t g t c g t c c g a c t t a t t a t t t t t t t a a t c t t t g a t a a t a

360 piggyBac right (3') inverted repeat t t a a c c c t a g a a a g a t a a t c a t a t t g t g a c g t a c g t t a a a g a t a a t c a t g c g t a a a a t t g a a t t g g g a t c t t t c t a t t a g t a t a a c a c t g c a t g c a a t t t c t a t t a g t a c g c a t t t t a a c

420 piggyBac right (3') inverted repeat a c g c a t g t g t t t t a t c g g t c t g t a t a t c g a g g t t t a t t t a t t a a t t t g a a t a g a t a t t a a t g c g t a c a c a a a a t a g c c a g a c a t a t a g c t c c a a a t a a a t a a t t a a a c t t a t c t a t a a t t

480 g t t t t a t t a t a t t t a c a c t t a c a t a c t a a t a a t a a a t t c a a c a a a c a a t t t a t t t a t g t t c a a a a t a a t a t a a a t g t g a a t g t a t g a t t a t t a t t t a a g t t g t t t g t t a a a t a a a t a c a a

540 t a t t t a t t t a t t a a a a a a a a a c a a a a a c t c a a a a t t t c t t c t a t a a a g t a a c a a a a c t t t a t a a a t a a a t a a t t t t t t t t t g t t t t t g a g t t t t a a a g a a g a t a t t t c a t t g t t t t g a a a

600 t a a a c a t t c t c t c t t t t a c a a a a a t a a a c t t a t t t t g t a c t t t a a a a a c a g t c a t g t t g t a t t t g t a a g a g a g a a a a t g t t t t t a t t t g a a t a a a a c a t g a a a t t t t t g t c a g t a c a a c a

660 a t t a t a a a a t a a g t a a t t a g c t t a a c t t a t a c a t a a t a g a a a c a a a t t a t a c t t a t t a g t t a a t a t t t t a t t c a t t a a t c g a a t t g a a t a t g t a t t a t c t t t g t t t a a t a t g a a t a a t c a

720 *FCSQCMDINHERCIVKKC c a g t c a g a a a c a a c t t t g g c a c a t a t c a a t a t t a t g c t c t c g a c a a a t a a c t t t t t t g c a g t c a g t c t t t g t t g a a a c c g t g t a t a g t t a t a a t a c g a g a g c t g t t t a t t g a a a a a a c g t

780 KKCSANAKRRIKSPCYTCYT t t t t t t g c a c g a t g c a t t t g c c t t t c g c c t t a t t t t a g a g g g g c a g t a a g t a c a g t a a g t a a a a a a c g t g c t a c g t a a a c g g a a a g c g g a a t a a a a t c t c c c c g t c a t t c a t g t c a t t c a

840 RKKMVPEETSDDSTGPVENP a c g t t t t t t c a t t a c t g g c t c t t c a g t a c t g t c a t c t g a t g t a c c a g g c a c t t c a t t t g g t g c a a a a a a g t a a t g a c c g a g a a g t c a t g a c a g t a g a c t a c a t g g t c c g t g a a g t a a a c c

900 LINSINDRLYRKLTPAELRK c a a a a t a t t a g a g a t a t t a t c g c g c a a a t a t c t c t t c a a a g t a g g a g c t t c t a a a c g c t t g t t t t a t a a t c t c t a t a a t a g c g c g t t t a t a g a g a a g t t t c a t c c t c g a a g a t t t g c g a a

960 RMFSSTLSMYLNRMFKKRSQ a c g c a t a a a c g a t g a c g t c a g g c t c a t g t a a a g g t t t c t c a t a a a t t t t t t g c g a c t t t g t g c g t a t t t g c t a c t g c a g t c c g a g t a c a t t t c c a a a g a g t a t t t a a a a a a c g c t g a a a c

1020 VKEGKSSVNHSYIIFSNICA a a c c t t t t c t c c c t t g c t a c t g a c a t t a t g g c t g t a t a t a a t a a a a g a a t t t a t g c a g g c t t g g a a a a g a g g g a a c g a t g a c t g t a a t a c c g a c a t a t a t t a t t t t c t t a a a t a c g t c c g

1080 INIMGYLLAMPWRNTKRSCT a a t g t t t a t c a t t c c g t a c a a t a a t g c c a t a g g c c a c c t a t t c g t c t t c c t a c t g c a g g t t t a c a a a t a g t a a g g c a t g t t a t t a c g g t a t c c g g t g g a t a a g c a g a a g g a t g a c g t c c a

1140 MVSCMQDLTDVGGKTQNYYM c a t c a c a g a a c a c a t t t g g t c t a g c g t g t c c a c t c c g c c t t t a g t t t g a t t a t a a t a c a t g t a g t g t c t t g t g t a a a c c a g a t c g c a c a g g t g a g g c g g a a a t c a a a c t a a t a t t a t g t a

AgeI 1200 VMQPKGTSENISADEDCSSL a a c c a t t t g c g g t t t a c c g g t a c t t t c g t t g a t a g a a g c a t c c t c a t c a c a a g a t g a t a a t t g g t a a a c g c c a a a t g g c c a t g a a a g c a a c t a t c t t c g t a g g a g t a g t g t t c t a c t a t t

1260 LYVMKAPKPKYSVLTLPGDF t a a g t a t a c c a t c t t a g c t g g c t t c g g t t t a t a t g a g a c g a g a g t a a g g g g t c c g t c a a a a t t c a t a t g g t a g a a t c g a c c g a a g c c a a a t a t a c t c t g c t c t c a t t c c c c a g g c a g t t t

BspDIClaI 1320 CFM a c a a a a c a t c g a t g t t c c c a c t g g c c t g g a g c g a c t g t t t t t c a g t a c t t c c g g t a t c t c t g t t t t g t a g c t a c a a g g g t g a c c g g a c c t c g c t g a c a a a a a g t c a t g a a g g c c a t a g a g

AvrII 1380 g c g t t t g t t t g a t c g c a c g g t t c c c a c a a t g g t t a a t t c g a g c t c g c c c g g g g t c c t a g g c g c a a a c a a a c t a g c g t g c c a a g g g t g t t a c c a a t t a a g c t c g a g c g g g c c c c a g g a t c c

SacII 1440 attB t c g a c g a t g t a g g t c a c g g t c t c g a a g c c g c g g t g c g g g t g c c a g g g c g t g c c c t t g g g c a g c t g c t a c a t c c a g t g c c a g a g c t t c g g c g c c a c g c c c a c g g t c c c g c a c g g g a a c c c g

1500 attB t c c c c g g g c g c g t a c t c c a c c t c a c c c a t c t g g t c c a t c a t g a t g a a c g g g t c g a g g t g g a g g g g c c c g c g c a t g a g g t g g a g t g g g t a g a c c a g g t a g t a c t a c t t g c c c a g c t c c a c c

1560 c g g t a g t t g a t c c c g g c g a a c g c g c g g c g c a c c g g g a a g c c c t c g c c c t c g a a a c c g c t g g c c a t c a a c t a g g g c c g c t t g c g c g c c g c g t g g c c c t t c g g g a g c g g g a g c t t t g g c g a c

Tth111IPflFI 1620 g g c g c g g t g g t c a c g g t g a g c a c g g g a c g t g c g a c g g c g t c g g c g g g t g c g g a t a c g c g g c c g c g c c a c c a g t g c c a c t c g t g c c c t g c a c g c t g c c g c a g c c g c c c a c g c c t a t g c g c c

PaeR7IXhoI 1680 g g c a g c g t c a g c g g g t t c t c g a c g g t c a c g g c g g g c a t g t c g a c t c g a g a a a t t t c t c t g c c g t c g c a g t c g c c c a a g a g c t g c c a g t g c c g c c c g t a c a g c t g a g c t c t t t a a a g a g a c

1740 g c c g t t a t t c g t t a t t c t c t c t t t t c t t t t t g g g t c t c t c c c t c t c t g c a c t a a t g c t c t c g g c a a t a a g c a a t a a g a g a g a a a a g a a a a a c c c a g a g a g g g a g a g a c g t g a t t a c g a g a

1800 c t c a c t c t g t c a c a c a g t a a a c g g c a t a c t g c t c t c g t t g g t t c g a g a g a g c g c g c c t c g g a g t g a g a c a g t g t g t c a t t t g c c g t a t g a c g a g a g c a a c c a a g c t c t c t c g c g c g g a g c

NruI 1860 hsp70 promoter a a t g t t c g c g a a a a g a g c g c c g g a g t a t a a a t a g a g g c g c t t c g t c t a c g g a g c g a c a a t t t a c a a g c g c t t t t c t c g c g g c c t c a t a t t t a t c t c c g c g a a g c a g a t g c c t c g c t g t t a

1920 hsp70 promoter t c a a t t c a a a c a a g c a a a g t g a a c a c g t c g c t a a g c g a a a g c t a a g c a a a t a a a c a a g c g a g t t a a g t t t g t t c g t t t c a c t t g t g c a g c g a t t c g c t t t c g a t t c g t t t a t t t g t t c g c

1980 hsp70 promoter c a g c t g a a c a a g c t a a a c a a t c t g c a g t a a a g t g c a a g t t a a a g t g a a t c a a t t a a a a g t g t c g a c t t g t t c g a t t t g t t a g a c g t c a t t t c a c g t t c a a t t t c a c t t a g t t a a t t t t c a

2040 hsp70 promoter G C A A C T A C T G A A A T C T G C C A A G pCasper-hs a a c c a g c a a c c a a g t a a a t c a a c t g c a a c t a c t g a a a t c t g c c a a g a a g t a a t t a t t g a a t t g g t c g t t g g t t c a t t t a g t t g a c g t t g a t g a c t t t a g a c g g t t c t t c a t t a a t a a c t t

PmeI 2100 1 5 10 DYKDHDGDYK hsp70 promoter 3xFLAG MDYKDHDGDYK *VFHVILVVVSVVL t a c a a g a a g a g a a c t c t g t t t a a a c a a a a t g g a c t a t a a g g a c c a c g a c g g a g a c t a c a a a t g t t c t t c t c t t g a g a c a a a t t t g t t t t a c c t g a t a t t c c t g g t g c t g c c t c t g a t g t t

2160 15 20 DHDIDYKDDDDK MA 130 PKKKRK 3xFLAG SV40 NLS DHDIDYKDDDDKMAPKKKRK IMINIVFVIVILHGWLLLPL g g a t c a t g a t a t t g a t t a c a a a g a c g a t g a c g a t a a g a t g g c c c c a a a g a a g a a g c g g a a c c t a g t a c t a t a a c t a a t g t t t c t g c t a c t g c t a t t c t a c c g g g g t t t c t t c t t c g c c t t

2220 5 10 DKKYSIGLDIG Cas9 V (in frame with SV40 NLS) GIHGVPAA 5 10 DKKYSIGLDIG SV40 NLS Cas9 VGIHGVPAADKKYSIGLDIG DTDVSDWCGVLLVADAQVDA g g t c g g t a t c c a c g g a g t c c c a g c a g c c g a c a a g a a g t a c a g c a t c g g c c t g g a c a t c g g c c a g c c a t a g g t g c c t c a g g g t c g t c g g c t g t t c t t c a t g t c g t a g c c g g a c c t g t a g c c

2280 15 20 25 30 TNSVGWAVITDEYKVPSKKF Cas9 15 20 25 30 TNSVGWAVITDEYKVPSKKF Cas9 TNSVGWAVITDEYKVPSKKF GVRHAPGHDGVLVLHGALFE c a c c a a c t c t g t g g g c t g g g c c g t g a t c a c c g a c g a g t a c a a g g t g c c c a g c a a g a a a t t g t g g t t g a g a c a c c c g a c c c g g c a c t a g t g g c t g c t c a t g t t c c a c g g g t c g t t c t t t a a

2340 35 40 45 50 KVLGNTDRHSIKKNLIGALL Cas9 35 40 45 50 KVLGNTDRHSIKKNLIGALL Cas9 KVLGNTDRHSIKKNLIGALL LHQAVGVPVADLLVQDSGQQ c a a g g t g c t g g g c a a c a c c g a c c g g c a c a g c a t c a a g a a g a a c c t g a t c g g a g c c c t g c t g t t c c a c g a c c c g t t g t g g c t g g c c g t g t c g t a g t t c t t c t t g g a c t a g c c t c g g g a c g a

2400 55 60 65 70 FDSGETAEATRLKRTARRRY Cas9 55 60 65 70 FDSGETAEATRLKRTARRRY Cas9 FDSGETAEATRLKRTARRRY EVAAFCGLGGPQLSGGSSSV g t t c g a c a g c g g c g a a a c a g c c g a g g c c a c c c g g c t g a a g a g a a c c g c c a g a a g a a g a t a c a a g c t g t c g c c g c t t t g t c g g c t c c g g t g g g c c g a c t t c t c t t g g c g g t c t t c t t c t a t

BglII 2460 75 80 85 90 TRRKNRICYLQEIFSNEMAK Cas9 75 80 85 90 TRRKNRICYLQEIFSNEMAK Cas9 TRRKNRICYLQEIFSNEMAK GSPLVPDAIQLLDEAVLHGL c a c c a g a c g g a a g a a c c g g a t c t g c t a t c t g c a a g a g a t c t t c a g c a a c g a g a t g g c c a a g t g g t c t g c c t t c t t g g c c t a g a c g a t a g a c g t t c t c t a g a a g t c g t t g c t c t a c c g g t t

2520 95 100 105 110 VDDSFFHRLEESFLVEEDKK Cas9 95 100 105 110 VDDSFFHRLEESFLVEEDKK Cas9 VDDSFFHRLEESFLVEEDKK HVVAEEVSQFLGEQHFLILL g g t g g a c g a c a g c t t c t t c c a c a g a c t g g a a g a g t c c t t c c t g g t g g a a g a g g a t a a g a a c c a c c t g c t g t c g a a g a a g g t g t c t g a c c t t c t c a g g a a g g a c c a c c t t c t c c t a t t c t t

2580 115 120 125 130 HERHPIFGNIVDEVAYHEKY Cas9 115 120 125 130 HERHPIFGNIVDEVAYHEKY Cas9 HERHPIFGNIVDEVAYHEKY VLPVGDEAVDHVLHGVVLLV g c a c g a g c g g c a c c c c a t c t t c g g c a a c a t c g t g g a c g a g g t g g c c t a c c a c g a g a a g t a c g t g c t c g c c g t g g g g t a g a a g c c g t t g t a g c a c c t g c t c c a c c g g a t g g t g c t c t t c a t

2640 135 140 145 150 PTIYHLRKKLVDSTDKADLR Cas9 135 140 145 150 PTIYHLRKKLVDSTDKADLR Cas9 PTIYHLRKKLVDSTDKADLR GGDVVQSLFQHVAGVLGVQP c c c c a c c a t c t a c c a c c t g a g a a a g a a a c t g g t g g a c a g c a c c g a c a a g g c c g a c c t g c g g g g g t g g t a g a t g g t g g a c t c t t t c t t t g a c c a c c t g t c g t g g c t g t t c c g g c t g g a c g c

2700 155 160 165 170 LIYLALAHMIKFRGHFLIEG Cas9 155 160 165 170 LIYLALAHMIKFRGHFLIEG Cas9 LIYLALAHMIKFRGHFLIEG QDIQGQGVHDLEPAVEQDLA g c t g a t c t a t c t g g c c c t g g c c c a c a t g a t c a a g t t c c g g g g c c a c t t c c t g a t c g a g g g c g a c t a g a t a g a c c g g g a c c g g g t g t a c t a g t t c a a g g c c c c g g t g a a g g a c t a g c t c c c

2760 175 180 185 190 DLNPDNSDVDKLFIQLVQTY Cas9 175 180 185 190 DLNPDNSDVDKLFIQLVQTY Cas9 DLNPDNSDVDKLFIQLVQTY VQVGVVAVHVLQEDLQHLGV c g a c c t g a a c c c c g a c a a c a g c g a c g t g g a c a a g c t g t t c a t c c a g c t g g t g c a g a c c t a g c t g g a c t t g g g g c t g t t g t c g c t g c a c c t g t t c g a c a a g t a g g t c g a c c a c g t c t g g a t

2820 195 200 205 210 NQLFEENPINASGVDAKAIL Cas9 195 200 205 210 NQLFEENPINASGVDAKAIL Cas9 NQLFEENPINASGVDAKAIL VLQELFVGDVGAAHVGLGDQ c a a c c a g c t g t t c g a g g a a a a c c c c a t c a a c g c c a g c g g c g t g g a c g c c a a g g c c a t c c t g t t g g t c g a c a a g c t c c t t t t g g g g t a g t t g c g g t c g c c g c a c c t g c g g t t c c g g t a g g a

2880 215 220 225 230 SARLSKSRRLENLIAQLPGE Cas9 215 220 225 230 SARLSKSRRLENLIAQLPGE Cas9 SARLSKSRRLENLIAQLPGE RGSQALASPQFIQDGLQGAL g t c t g c c a g a c t g a g c a a g a g c a g a c g g c t g g a a a a t c t g a t c g c c c a g c t g c c c g g c g a c a g a c g g t c t g a c t c g t t c t c g t c t g c c g a c c t t t t a g a c t a g c g g g t c g a c g g g c c g c t

2940 235 240 245 250 KKNGLFGNLIALSLGLTPNF Cas9 235 240 245 250 KKNGLFGNLIALSLGLTPNF Cas9 KKNGLFGNLIALSLGLTPNF LLIAQESVQNGQAQAQGGVE g a a g a a g a a t g g c c t g t t c g g a a a c c t g a t t g c c c t g a g c c t g g g c c t g a c c c c c a a c t t c t t c t t c t t a c c g g a c a a g c c t t t g g a c t a a c g g g a c t c g g a c c c g g a c t g g g g g t t g a a

3000 255 260 265 270 KSNFDLAEDAKLQLSKDTYD Cas9 255 260 265 270 KSNFDLAEDAKLQLSKDTYD Cas9 KSNFDLAEDAKLQLSKDTYD LAVEVQGLIGFQLQALVGVV c a a g a g c a a c t t c g a c c t g g c c g a g g a t g c c a a a c t g c a g c t g a g c a a g g a c a c c t a c g a g t t c t c g t t g a a g c t g g a c c g g c t c c t a c g g t t t g a c g t c g a c t c g t t c c t g t g g a t g c t

3060 275 280 285 290 DDLDNLLAQIGDQYADLFLA Cas9 275 280 285 290 DDLDNLLAQIGDQYADLFLA Cas9 DDLDNLLAQIGDQYADLFLA VVQVVQQGLDAVLVGVQKQG c g a c g a c c t g g a c a a c c t g c t g g c c c a g a t c g g c g a c c a g t a c g c c g a c c t g t t t c t g g c g c t g c t g g a c c t g t t g g a c g a c c g g g t c t a g c c g c t g g t c a t g c g g c t g g a c a a a g a c c g

3120 295 300 305 310 AKNLSDAILLSDILRVNTEI Cas9 295 300 305 310 AKNLSDAILLSDILRVNTEI Cas9 AKNLSDAILLSDILRVNTEI GLVQGVGDQQAVDQSHVGLD c g c c a a g a a c c t g t c c g a c g c c a t c c t g c t g a g c g a c a t c c t g a g a g t g a a c a c c g a g a t g c g g t t c t t g g a c a g g c t g c g g t a g g a c g a c t c g c t g t a g g a c t c t c a c t t g t g g c t c t a

3180 315 320 325 330 TKAPLSASMIKRYDEHHQDL Cas9 315 320 325 330 TKAPLSASMIKRYDEHHQDL Cas9 TKAPLSASMIKRYDEHHQDL GLGGQAGRHDLSVVLVVLVQ c a c c a a g g c c c c c c t g a g c g c c t c t a t g a t c a a g a g a t a c g a c g a g c a c c a c c a g g a c c t g t g g t t c c g g g g g g a c t c g c g g a g a t a c t a g t t c t c t a t g c t g c t c g t g g t g g t c c t g g a

3240 335 340 345 350 TLLKALVRQQLPEKYKEIFF Cas9 335 340 345 350 TLLKALVRQQLPEKYKEIFF Cas9 TLLKALVRQQLPEKYKEIFF GQQFSEHPLLQRLLVFLNEE g a c c c t g c t g a a a g c t c t c g t g c g g c a g c a g c t g c c t g a g a a g t a c a a a g a g a t t t t c t t c t g g g a c g a c t t t c g a g a g c a c g c c g t c g t c g a c g g a c t c t t c a t g t t t c t c t a a a a g a a

3300 355 360 365 370 DQSKNGYAGYIDGGASQEEF Cas9 355 360 365 370 DQSKNGYAGYIDGGASQEEF Cas9 DQSKNGYAGYIDGGASQEEF VLALVAVGAVNVASGALFLE c g a c c a g a g c a a g a a c g g c t a c g c c g g c t a c a t t g a c g g c g g a g c c a g c c a g g a a g a g t t g c t g g t c t c g t t c t t g c c g a t g c g g c c g a t g t a a c t g c c g c c t c g g t c g g t c c t t c t c a a

3360 375 380 385 390 YKFIKPILEKMDGTEELLVK Cas9 375 380 385 390 YKFIKPILEKMDGTEELLVK Cas9 YKFIKPILEKMDGTEELLVK VLEDLGDQFLHVAGLFQEHL c t a c a a g t t c a t c a a g c c c a t c c t g g a a a a g a t g g a c g g c a c c g a g g a a c t g c t c g t g a a g a t g t t c a a g t a g t t c g g g t a g g a c c t t t t c t a c c t g c c g t g g c t c c t t g a c g a g c a c t t

3420 395 400 405 410 LNREDLLRKQRTFDNGSIPH Cas9 395 400 405 410 LNREDLLRKQRTFDNGSIPH Cas9 LNREDLLRKQRTFDNGSIPH QVSLVQQPLLPGEVVAADGV g c t g a a c a g a g a g g a c c t g c t g c g g a a g c a g c g g a c c t t c g a c a a c g g c a g c a t c c c c c a c g a c t t g t c t c t c c t g g a c g a c g c c t t c g t c g c c t g g a a g c t g t t g c c g t c g t a g g g g g t

3480 415 420 425 430 QIHLGELHAILRRQEDFYPF Cas9 415 420 425 430 QIHLGELHAILRRQEDFYPF Cas9 QIHLGELHAILRRQEDFYPF LDVQSLQVGNQPPLFIKVWE c c a g a t c c a c c t g g g a g a g c t g c a c g c c a t t c t g c g g c g g c a g g a a g a t t t t t a c c c a t t g g t c t a g g t g g a c c c t c t c g a c g t g c g g t a a g a c g c c g c c g t c c t t c t a a a a a t g g g t a a

3540 435 440 445 450 LKDNREKIEKILTFRIPYYV Cas9 435 440 445 450 LKDNREKIEKILTFRIPYYV Cas9 LKDNREKIEKILTFRIPYYV QLVVPFLDLLDQGEADGVVH c c t g a a g g a c a a c c g g g a a a a g a t c g a g a a g a t c c t g a c c t t c c g c a t c c c c t a c t a c g t g g a c t t c c t g t t g g c c c t t t t c t a g c t c t t c t a g g a c t g g a a g g c g t a g g g g a t g a t g c a

3600 455 460 465 470 GPLARGNSRFAWMTRKSEET Cas9 455 460 465 470 GPLARGNSRFAWMTRKSEET Cas9 GPLARGNSRFAWMTRKSEET ARQGPSVASEGPHGSLALFG g g g c c c t c t g g c c a g g g g a a a c a g c a g a t t c g c c t g g a t g a c c a g a a a g a g c g a g g a a a c c c c g g g a g a c c g g t c c c c t t t g t c g t c t a a g c g g a c c t a c t g g t c t t t c t c g c t c c t t t g

3660 475 480 485 490 ITPWNFEEVVDKGASAQSFI Cas9 475 480 485 490 ITPWNFEEVVDKGASAQSFI Cas9 ITPWNFEEVVDKGASAQSFI DGGPVELFHHVLASGGLAED c a t c a c c c c c t g g a a c t t c g a g g a a g t g g t g g a c a a g g g c g c t t c c g c c c a g a g c t t c a t g t a g t g g g g g a c c t t g a a g c t c c t t c a c c a c c t g t t c c c g c g a a g g c g g g t c t c g a a g t a

3720 495 500 505 510 ERMTNFDKNLPNEKVLPKHS Cas9 495 500 505 510 ERMTNFDKNLPNEKVLPKHS Cas9 ERMTNFDKNLPNEKVLPKHS LPHGVEILVQGVLLHQGLVA c g a g c g g a t g a c c a a c t t c g a t a a g a a c c t g c c c a a c g a g a a g g t g c t g c c c a a g c a c a g g c t c g c c t a c t g g t t g a a g c t a t t c t t g g a c g g g t t g c t c t t c c a c g a c g g g t t c g t g t c

3780 515 520 525 530 LLYEYFTVYNELTKVKYVTE Cas9 515 520 525 530 LLYEYFTVYNELTKVKYVTE Cas9 LLYEYFTVYNELTKVKYVTE QQVLVEGHIVLQGFHFVHGL c c t g c t g t a c g a g t a c t t c a c c g t g t a t a a c g a g c t g a c c a a a g t g a a a t a c g t g a c c g a g g a c g a c a t g c t c a t g a a g t g g c a c a t a t t g c t c g a c t g g t t t c a c t t t a t g c a c t g g c t

3840 535 540 545 550 GMRKPAFLSGEQKKAIVDLL Cas9 535 540 545 550 GMRKPAFLSGEQKKAIVDLL Cas9 GMRKPAFLSGEQKKAIVDLL SHSLGGEQAALLFLGDHVQQ g g g a a t g a g a a a g c c c g c c t t c c t g a g c g g c g a g c a g a a a a a g g c c a t c g t g g a c c t g c t c c c t t a c t c t t t c g g g c g g a a g g a c t c g c c g c t c g t c t t t t t c c g g t a g c a c c t g g a c g a

3900 555 560 565 570 FKTNRKVTVKQLKEDYFKKI Cas9 555 560 565 570 FKTNRKVTVKQLKEDYFKKI Cas9 FKTNRKVTVKQLKEDYFKKI ELGVPFHGHLLQFLVVELFD g t t c a a g a c c a a c c g g a a a g t g a c c g t g a a g c a g c t g a a a g a g g a c t a c t t c a a g a a a a t c a a g t t c t g g t t g g c c t t t c a c t g g c a c t t c g t c g a c t t t c t c c t g a t g a a g t t c t t t t a

PasI 3960 575 580 585 590 ECFDSVEISGVEDRFNASLG Cas9 575 580 585 590 ECFDSVEISGVEDRFNASLG Cas9 ECFDSVEISGVEDRFNASLG LAEVGHFDGAHFIPEVGGQA c g a g t g c t t c g a c t c c g t g g a a a t c t c c g g c g t g g a a g a t c g g t t c a a c g c c t c c c t g g g g c t c a c g a a g c t g a g g c a c c t t t a g a g g c c g c a c c t t c t a g c c a a g t t g c g g a g g g a c c c

4020 595 600 605 610 TYHDLLKIIKDKDFLDNEEN Cas9 595 600 605 610 TYHDLLKIIKDKDFLDNEEN Cas9 TYHDLLKIIKDKDFLDNEEN CVVIQQFNDLVLVEQVILFV c a c a t a c c a c g a t c t g c t g a a a a t t a t c a a g g a c a a g g a c t t c c t g g a c a a t g a g g a a a a g t g t a t g g t g c t a g a c g a c t t t t a a t a g t t c c t g t t c c t g a a g g a c c t g t t a c t c c t t t t

4080 615 620 625 630 EDILEDIVLTLTLFEDREMI Cas9 615 620 625 630 EDILEDIVLTLTLFEDREMI Cas9 EDILEDIVLTLTLFEDREMI LVNQFIDHQGQCQKLVSLHD c g a g g a c a t t c t g g a a g a t a t c g t g c t g a c c c t g a c a c t g t t t g a g g a c a g a g a g a t g a t g c t c c t g t a a g a c c t t c t a t a g c a c g a c t g g g a c t g t g a c a a a c t c c t g t c t c t c t a c t a

4140 635 640 645 650 EERLKTYAHLFDDKVMKQLK Cas9 635 640 645 650 EERLKTYAHLFDDKVMKQLK Cas9 EERLKTYAHLFDDKVMKQLK *AWRNSSLTIFCSF LFPQFGIGVQEVVFHHLLQL c g a g g a a c g g c t g a a a a c c t a t g c c c a c c t g t t c g a c g a c a a a g t g a t g a a g c a g c t g a a g c t c c t t g c c g a c t t t t g g a t a c g g g t g g a c a a g c t g c t g t t t c a c t a c t t c g t c g a c t t

4200 655 660 665 670 RRRYTGWGRLSRKLINGIRD Cas9 655 660 665 670 RRRYTGWGRLSRKLINGIRD Cas9 RRRYTGWGRLSRKLINGIRD RRLYVPQPLSLRFSILPMRS PPSVGAPAPQAPLQDVADPV g c g g c g g a g a t a c a c c g g c t g g g g c a g g c t g a g c c g g a a g c t g a t c a a c g g c a t c c g g g a c g c c g c c t c t a t g t g g c c g a c c c c g t c c g a c t c g g c c t t c g a c t a g t t g c c g t a g g c c c t

4260 675 680 685 690 KQSGKTILDFLKSDGFANRN Cas9 675 680 685 690 KQSGKTILDFLKSDGFANRN Cas9 KQSGKTILDFLKSDGFANRN LCDPLVIRSKRFDSPKALLF LLGALCDQIEQLGVAEGVSV c a a g c a g t c c g g c a a g a c a a t c c t g g a t t t c c t g a a g t c c g a c g g c t t c g c c a a c a g a a a g t t c g t c a g g c c g t t c t g t t a g g a c c t a a a g g a c t t c a g g c t g c c g a a g c g g t t g t c t t t

4320 695 700 705 710 FMQLIHDDSLTFKEDIQKAQ Cas9 695 700 705 710 FMQLIHDDSLTFKEDIQKAQ Cas9 FMQLIHDDSLTFKEDIQKAQ KMCSIWSSLRVKLSSMWFAW EHLQDVVVAQGKFLVDLFGL c t t c a t g c a g c t g a t c c a c g a c g a c a g c c t g a c c t t t a a a g a g g a c a t c c a g a a a g c c c a g a a g t a c g t c g a c t a g g t g c t g c t g t c g g a c t g g a a a t t t c t c c t g t a g g t c t t t c g g g t

4380 715 720 725 730 VSGQGDSLHEHIANLAGSPA Cas9 715 720 725 730 VSGQGDSLHEHIANLAGSPA Cas9 VSGQGDSLHEHIANLAGSPA TDPWPSLRCSCMALRAPLGA HGALAIAQVLVNGIQGAAGG g g t g t c c g g c c a g g g c g a t a g c c t g c a c g a g c a c a t t g c c a a t c t g g c c g g c a g c c c c g c c c a c a g g c c g g t c c c g c t a t c g g a c g t g c t c g t g t a a c g g t t a g a c c g g c c g t c g g g g c g

4440 735 740 745 750 IKKGILQTVKVVDELVKVMG Cas9 735 740 745 750 IKKGILQTVKVVDELVKVMG Cas9 IKKGILQTVKVVDELVKVMG MLFPMRCVTFTTSSSTFTIP NLLADQLCHLHHVLEHFHHA c a t t a a g a a g g g c a t c c t g c a g a c a g t g a a g g t g g t g g a c g a g c t c g t g a a a g t g a t g g g g t a a t t c t t c c c g t a g g a c g t c t g t c a c t t c c a c c a c c t g c t c g a g c a c t t t c a c t a c c c

4500 755 760 765 770 RHKPENIVIEMARENQTTQK Cas9 755 760 765 770 RHKPENIVIEMARENQTTQK Cas9 RHKPENIVIEMARENQTTQK RCLGSFMTISIALSFWVVWF PVLGLVDHDFHGSLVLGGLL c c g g c a c a a g c c c g a g a a c a t c g t g a t c g a a a t g g c c a g a g a g a a c c a g a c c a c c c a g a a g g c c g t g t t c g g g c t c t t g t a g c a c t a g c t t t a c c g g t c t c t c t t g g t c t g g t g g g t c t t

4560 775 780 785 790 GQKNSRERMKRIEEGIKELG Cas9 775 780 785 790 GQKNSRERMKRIEEGIKELG Cas9 GQKNSRERMKRIEEGIKELG PCFFLRSLIFRISSPM SLLVAALSHLPDFLADFLQA g g g a c a g a a g a a c a g c c g c g a g a g a a t g a a g c g g a t c g a a g a g g g c a t c a a a g a g c t g g g c c c t g t c t t c t t g t c g g c g c t c t c t t a c t t c g c c t a g c t t c t c c c g t a g t t t c t c g a c c c

4620 795 800 805 810 SQILKEHPVENTQLQNEKLY Cas9 795 800 805 810 SQILKEHPVENTQLQNEKLY Cas9 SQILKEHPVENTQLQNEKLY ALDQFFVGHFVGLQLVLLQV c a g c c a g a t c c t g a a a g a a c a c c c c g t g g a a a a c a c c c a g c t g c a g a a c g a g a a g c t g t a g t c g g t c t a g g a c t t t c t t g t g g g g c a c c t t t t g t g g g t c g a c g t c t t g c t c t t c g a c a t

4680 815 820 825 830 LYYLQNGRDMYVDQELDINR Cas9 815 820 825 830 LYYLQNGRDMYVDQELDINR Cas9 LYYLQNGRDMYVDQELDINR QVVQLIPPIHVHVLFQVDVP c c t g t a c t a c c t g c a g a a t g g g c g g g a t a t g t a c g t g g a c c a g g a a c t g g a c a t c a a c c g g g a c a t g a t g g a c g t c t t a c c c g c c c t a t a c a t g c a c c t g g t c c t t g a c c t g t a g t t g g c

4740 835 840 845 850 LSDYDVDHIVPQSFLKDDSI Cas9 835 840 845 850 LSDYDVDHIVPQSFLKDDSI Cas9 LSDYDVDHIVPQSFLKDDSI QGVVIHVM g c t g t c c g a c t a c g a t g t g g a c c a t a t c g t g c c t c a g a g c t t t c t g a a g g a c g a c t c c a t c g a c a g g c t g a t g c t a c a c c t g g t a t a g c a c g g a g t c t c g a a a g a c t t c c t g c t g a g g t a

4800 855 860 865 870 DNKVLTRSDKNRGKSDNVPS Cas9 855 860 865 870 DNKVLTRSDKNRGKSDNVPS Cas9 DNKVLTRSDKNRGKSDNVPS c g a c a a c a a g g t g c t g a c c a g a a g c g a c a a g a a c c g g g g c a a g a g c g a c a a c g t g c c c t c g c t g t t g t t c c a c g a c t g g t c t t c g c t g t t c t t g g c c c c g t t c t c g c t g t t g c a c g g g a g

4860 875 880 885 890 EEVVKKMKNYWRQLLNAKLI Cas9 875 880 885 890 EEVVKKMKNYWRQLLNAKLI Cas9 EEVVKKMKNYWRQLLNAKLI c g a a g a g g t c g t g a a g a a g a t g a a g a a c t a c t g g c g g c a g c t g c t g a a c g c c a a g c t g a t g c t t c t c c a g c a c t t c t t c t a c t t c t t g a t g a c c g c c g t c g a c g a c t t g c g g t t c g a c t a

4920 895 900 905 910 TQRKFDNLTKAERGGLSELD Cas9 895 900 905 910 TQRKFDNLTKAERGGLSELD Cas9 TQRKFDNLTKAERGGLSELD t a c c c a g a g a a a g t t c g a c a a t c t g a c c a a g g c c g a g a g a g g c g g c c t g a g c g a a c t g g a a t g g g t c t c t t t c a a g c t g t t a g a c t g g t t c c g g c t c t c t c c g c c g g a c t c g c t t g a c c t

PmlI 4980 915 920 925 930 KAGFIKRQLVETRQITKHVA Cas9 915 920 925 930 KAGFIKRQLVETRQITKHVA Cas9 KAGFIKRQLVETRQITKHVA t a a g g c c g g c t t c a t c a a g a g a c a g c t g g t g g a a a c c c g g c a g a t c a c a a a g c a c g t g g c a t t c c g g c c g a a g t a g t t c t c t g t c g a c c a c c t t t g g g c c g t c t a g t g t t t c g t g c a c c g

5040 935 940 945 950 QILDSRMNTKYDENDKLIRE Cas9 935 940 945 950 QILDSRMNTKYDENDKLIRE Cas9 QILDSRMNTKYDENDKLIRE a c a g a t c c t g g a c t c c c g g a t g a a c a c t a a g t a c g a c g a g a a t g a c a a g c t g a t c c g g g a t g t c t a g g a c c t g a g g g c c t a c t t g t g a t t c a t g c t g c t c t t a c t g t t c g a c t a g g c c c t

5100 955 960 965 970 VKVITLKSKLVSDFRKDFQF Cas9 955 960 965 970 VKVITLKSKLVSDFRKDFQF Cas9 VKVITLKSKLVSDFRKDFQF a g t g a a a g t g a t c a c c c t g a a g t c c a a g c t g g t g t c c g a t t t c c g g a a g g a t t t c c a g t t t c a c t t t c a c t a g t g g g a c t t c a g g t t c g a c c a c a g g c t a a a g g c c t t c c t a a a g g t c a a

5160 975 980 985 990 YKVREINNYHHAHDAYLNAV Cas9 975 980 985 990 YKVREINNYHHAHDAYLNAV Cas9 YKVREINNYHHAHDAYLNAV t t a c a a a g t g c g c g a g a t c a a c a a c t a c c a c c a c g c c c a c g a c g c c t a c c t g a a c g c c g t a a t g t t t c a c g c g c t c t a g t t g t t g a t g g t g g t g c g g g t g c t g c g g a t g g a c t t g c g g c a

5220 995 1000 1005 1010 VGTALIKKYPKLESEFVYGD Cas9 995 1000 1005 1010 VGTALIKKYPKLESEFVYGD Cas9 VGTALIKKYPKLESEFVYGD c g t g g g a a c c g c c c t g a t c a a a a a g t a c c c t a a g c t g g a a a g c g a g t t c g t g t a c g g c g a g c a c c c t t g g c g g g a c t a g t t t t t c a t g g g a t t c g a c c t t t c g c t c a a g c a c a t g c c g c t

5280 1015 1020 1025 1030 YKVYDVRKMIAKSEQEIGKA Cas9 1015 1020 1025 1030 YKVYDVRKMIAKSEQEIGKA Cas9 YKVYDVRKMIAKSEQEIGKA c t a c a a g g t g t a c g a c g t g c g g a a g a t g a t c g c c a a g a g c g a g c a g g a a a t c g g c a a g g c g a t g t t c c a c a t g c t g c a c g c c t t c t a c t a g c g g t t c t c g c t c g t c c t t t a g c c g t t c c g

5340 1035 1040 1045 1050 TAKYFFYSNIMNFFKTEITL Cas9 1035 1040 1045 1050 TAKYFFYSNIMNFFKTEITL Cas9 TAKYFFYSNIMNFFKTEITL t a c c g c c a a g t a c t t c t t c t a c a g c a a c a t c a t g a a c t t t t t c a a g a c c g a g a t t a c c c t a t g g c g g t t c a t g a a g a a g a t g t c g t t g t a g t a c t t g a a a a a g t t c t g g c t c t a a t g g g a

5400 1055 1060 1065 1070 ANGEIRKRPLIETNGETGEI Cas9 1055 1060 1065 1070 ANGEIRKRPLIETNGETGEI Cas9 ANGEIRKRPLIETNGETGEI g g c c a a c g g c g a g a t c c g g a a g c g g c c t c t g a t c g a g a c a a a c g g c g a a a c c g g g g a g a t c c g g t t g c c g c t c t a g g c c t t c g c c g g a g a c t a g c t c t g t t t g c c g c t t t g g c c c c t c t a

5460 1075 1080 1085 1090 VWDKGRDFATVRKVLSMPQV Cas9 1075 1080 1085 1090 VWDKGRDFATVRKVLSMPQV Cas9 VWDKGRDFATVRKVLSMPQV c g t g t g g g a t a a g g g c c g g g a t t t t g c c a c c g t g c g g a a a g t g c t g a g c a t g c c c c a a g t g c a c a c c c t a t t c c c g g c c c t a a a a c g g t g g c a c g c c t t t c a c g a c t c g t a c g g g g t t c a

5520 1095 1100 1105 1110 NIVKKTEVQTGGFSKESILP Cas9 1095 1100 1105 1110 NIVKKTEVQTGGFSKESILP Cas9 NIVKKTEVQTGGFSKESILP g a a t a t c g t g a a a a a g a c c g a g g t g c a g a c a g g c g g c t t c a g c a a a g a g t c t a t c c t g c c c t t a t a g c a c t t t t t c t g g c t c c a c g t c t g t c c g c c g a a g t c g t t t c t c a g a t a g g a c g g

5580 1115 1120 1125 1130 KRNSDKLIARKKDWDPKKYG Cas9 1115 1120 1125 1130 KRNSDKLIARKKDWDPKKYG Cas9 KRNSDKLIARKKDWDPKKYG c a a g a g g a a c a g c g a t a a g c t g a t c g c c a g a a a g a a g g a c t g g g a c c c t a a g a a g t a c g g g t t c t c c t t g t c g c t a t t c g a c t a g c g g t c t t t c t t c c t g a c c c t g g g a t t c t t c a t g c c

5640 1135 1140 1145 1150 GFDSPTVAYSVLVVAKVEKG Cas9 1135 1140 1145 1150 GFDSPTVAYSVLVVAKVEKG Cas9 GFDSPTVAYSVLVVAKVEKG c g g c t t c g a c a g c c c c a c c g t g g c c t a t t c t g t g c t g g t g g t g g c c a a a g t g g a a a a g g g g c c g a a g c t g t c g g g g t g g c a c c g g a t a a g a c a c g a c c a c c a c c g g t t t c a c c t t t t c c c

5700 1155 1160 1165 1170 KSKKLKSVKELLGITIMERS Cas9 1155 1160 1165 1170 KSKKLKSVKELLGITIMERS Cas9 KSKKLKSVKELLGITIMERS c a a g t c c a a g a a a c t g a a g a g t g t g a a a g a g c t g c t g g g g a t c a c c a t c a t g g a a a g a a g g t t c a g g t t c t t t g a c t t c t c a c a c t t t c t c g a c g a c c c c t a g t g g t a g t a c c t t t c t t c

5760 1175 1180 1185 1190 SFEKNPIDFLEAKGYKEVKK Cas9 1175 1180 1185 1190 SFEKNPIDFLEAKGYKEVKK Cas9 SFEKNPIDFLEAKGYKEVKK c a g c t t c g a g a a g a a t c c c a t c g a c t t t c t g g a a g c c a a g g g c t a c a a a g a a g t g a a a a a g t c g a a g c t c t t c t t a g g g t a g c t g a a a g a c c t t c g g t t c c c g a t g t t t c t t c a c t t t t t

5820 1195 1200 1205 1210 DLIIKLPKYSLFELENGRKR Cas9 1195 1200 1205 1210 DLIIKLPKYSLFELENGRKR Cas9 DLIIKLPKYSLFELENGRKR g g a c c t g a t c a t c a a g c t g c c t a a g t a c t c c c t g t t c g a g c t g g a a a a c g g c c g g a a g a g c c t g g a c t a g t a g t t c g a c g g a t t c a t g a g g g a c a a g c t c g a c c t t t t g c c g g c c t t c t c

5880 1215 1220 1225 1230 MLASAGELQKGNELALPSKY Cas9 1215 1220 1225 1230 MLASAGELQKGNELALPSKY Cas9 MLASAGELQKGNELALPSKY a a t g c t g g c c t c t g c c g g c g a a c t g c a g a a g g g a a a c g a a c t g g c c c t g c c c t c c a a a t a t t a c g a c c g g a g a c g g c c g c t t g a c g t c t t c c c t t t g c t t g a c c g g g a c g g g a g g t t t a t

5940 1235 1240 1245 1250 VNFLYLASHYEKLKGSPEDN Cas9 1235 1240 1245 1250 VNFLYLASHYEKLKGSPEDN Cas9 VNFLYLASHYEKLKGSPEDN t g t g a a c t t c c t g t a c c t g g c c a g c c a c t a t g a g a a g c t g a a g g g c t c c c c c g a g g a t a a a c a c t t g a a g g a c a t g g a c c g g t c g g t g a t a c t c t t c g a c t t c c c g a g g g g g c t c c t a t t

6000 1255 1260 1265 1270 EQKQLFVEQHKHYLDEIIEQ Cas9 1255 1260 1265 1270 EQKQLFVEQHKHYLDEIIEQ Cas9 EQKQLFVEQHKHYLDEIIEQ t g a g c a g a a a c a g c t g t t t g t g g a a c a g c a c a a g c a c t a c c t g g a c g a g a t c a t c g a g c a a c t c g t c t t t g t c g a c a a a c a c c t t g t c g t g t t c g t g a t g g a c c t g c t c t a g t a g c t c g t

6060 1275 1280 1285 1290 ISEFSKRVILADANLDKVLS Cas9 1275 1280 1285 1290 ISEFSKRVILADANLDKVLS Cas9 ISEFSKRVILADANLDKVLS g a t c a g c g a g t t c t c c a a g a g a g t g a t c c t g g c c g a c g c t a a t c t g g a c a a a g t g c t g t c c t a g t c g c t c a a g a g g t t c t c t c a c t a g g a c c g g c t g c g a t t a g a c c t g t t t c a c g a c a g

6120 1295 1300 1305 1310 AYNKHRDKPIREQAENIIHL Cas9 1295 1300 1305 1310 AYNKHRDKPIREQAENIIHL Cas9 AYNKHRDKPIREQAENIIHL c g c c t a c a a c a a g c a c c g g g a t a a g c c c a t c a g a g a g c a g g c c g a g a a t a t c a t c c a c c t g c g g a t g t t g t t c g t g g c c c t a t t c g g g t a g t c t c t c g t c c g g c t c t t a t a g t a g g t g g a

6180 1315 1320 1325 1330 FTLTNLGAPAAFKYFDTTID Cas9 1315 1320 1325 1330 FTLTNLGAPAAFKYFDTTID Cas9 FTLTNLGAPAAFKYFDTTID g t t t a c c c t g a c c a a t c t g g g a g c c c c t g c c g c c t t c a a g t a c t t t g a c a c c a c c a t c g a c a a a t g g g a c t g g t t a g a c c c t c g g g g a c g g c g g a a g t t c a t g a a a c t g t g g t g g t a g c t

6240 1335 1340 1345 1350 RKRYTSTKEVLDATLIHQSI Cas9 1335 1340 1345 1350 RKRYTSTKEVLDATLIHQSI Cas9 RKRYTSTKEVLDATLIHQSI c c g g a a g a g g t a c a c c a g c a c c a a a g a g g t g c t g g a c g c c a c c c t g a t c c a c c a g a g c a t g g c c t t c t c c a t g t g g t c g t g g t t t c t c c a c g a c c t g c g g t g g g a c t a g g t g g t c t c g t a

6300 1355 1360 1365 TGLYETRIDLSQLGGD KRPA Cas9 1355 1360 1365 TGLYETRIDLSQLGGD 1 KRPA Cas9 nucleoplasmin NLS TGLYETRIDLSQLGGDKRPA c a c c g g c c t g t a c g a g a c a c g g a t c g a c c t g t c t c a g c t g g g a g g c g a c a a a a g g c c g g c g t g g c c g g a c a t g c t c t g t g c c t a g c t g g a c a g a g t c g a c c c t c c g c t g t t t t c c g g c c g

6360 (in frame with Cas9) ATKKAGQAKKKKEGRGSLLT 5 10 15 ATKKAGQAKKKK 140 145 EGRGSLLT nucleoplasmin NLS T2A ATKKAGQAKKKKEGRGSLLT g g c c a c g a a a a a g g c c g g c c a g g c a a a a a a g a a a a a g g a g g g c a g a g g a a g t c t t c t a a c c c g g t g c t t t t t c c g g c c g g t c c g t t t t t t c t t t t t c c t c c c g t c t c c t t c a g a a g a t t g

6420 (in frame with Cas9) CGDVEENPGPMVSKGEELFT 150 155 CGDVEENPGP 1 M 1a V 5 SKGEELFT T2A EGFP CGDVEENPGPMVSKGEELFT a t g c g g t g a c g t g g a g g a g a a t c c c g g c c c t a t g g t g a g c a a g g g c g a g g a g c t g t t c a c t a c g c c a c t g c a c c t c c t c t t a g g g c c g g g a t a c c a c t c g t t c c c g c t c c t c g a c a a g t g

6480 (in frame with Cas9) GVVPILVELDGDVNGHKFSV 10 15 20 25 GVVPILVELDGDVNGHKFSV EGFP GVVPILVELDGDVNGHKFSV c g g g g t g g t g c c c a t c c t g g t c g a g c t g g a c g g c g a c g t a a a c g g c c a c a a g t t c a g c g t g c c c c a c c a c g g g t a g g a c c a g c t c g a c c t g c c g c t g c a t t t g c c g g t g t t c a a g t c g c a

6540 (in frame with Cas9) SGEGEGDATYGKLTLKFICT 30 35 40 45 SGEGEGDATYGKLTLKFICT EGFP SGEGEGDATYGKLTLKFICT g t c c g g c g a g g g c g a g g g c g a t g c c a c c t a c g g c a a g c t g a c c c t g a a g t t c a t c t g c a c c a g g c c g c t c c c g c t c c c g c t a c g g t g g a t g c c g t t c g a c t g g g a c t t c a a g t a g a c g t g

6600 (in frame with Cas9) TSKLPVPWPTLVTTLTYGVQ 50 55 60 65 TSKLPVPWPTLVTTLTYGVQ EGFP TSKLPVPWPTLVTTLTYGVQ c a c c a g c a a g c t g c c c g t g c c c t g g c c c a c c c t c g t g a c c a c c c t g a c c t a c g g c g t g c a g t g g t c g t t c g a c g g g c a c g g g a c c g g g t g g g a g c a c t g g t g g g a c t g g a t g c c g c a c g t

6660 (in frame with Cas9) CFSRYPDHMKQHDFFKSAMP 70 75 80 85 CFSRYPDHMKQHDFFKSAMP EGFP CFSRYPDHMKQHDFFKSAMP g t g c t t c a g c c g c t a c c c c g a c c a c a t g a a g c a g c a c g a c t t c t t c a a g t c c g c c a t g c c c a c g a a g t c g g c g a t g g g g c t g g t g t a c t t c g t c g t g c t g a a g a a g t t c a g g c g g t a c g g

6720 (in frame with Cas9) EGYVQERTIFFKDDGNYKTR 90 95 100 105 EGYVQERTIFFKDDGNYKTR EGFP C T G C T G C C G T T G A T G T T C T G EXFP-R EGYVQERTIFFKDDGNYKTR c g a a g g c t a c g t c c a g g a g c g c a c c a t c t t c t t c a a g g a c g a c g g c a a c t a c a a g a c c c g g c t t c c g a t g c a g g t c c t c g c g t g g t a g a a g a a g t t c c t g c t g c c g t t g a t g t t c t g g g c

6780 (in frame with Cas9) AEVKFEGDTLVNRIELKGID 110 115 120 125 AEVKFEGDTLVNRIELKGID EGFP AEVKFEGDTLVNRIELKGID c g c c g a g g t g a a g t t c g a g g g c g a c a c c c t g g t g a a c c g c a t c g a g c t g a a g g g c a t c g a g c g g c t c c a c t t c a a g c t c c c g c t g t g g g a c c a c t t g g c g t a g c t c g a c t t c c c g t a g c t

6840 (in frame with Cas9) FKEDGNILGHKLEYNYNSHN 130 135 140 145 FKEDGNILGHKLEYNYNSHN EGFP FKEDGNILGHKLEYNYNSHN c t t c a a g g a g g a c g g c a a c a t c c t g g g g c a c a a g c t g g a g t a c a a c t a c a a c a g c c a c a a g a a g t t c c t c c t g c c g t t g t a g g a c c c c g t g t t c g a c c t c a t g t t g a t g t t g t c g g t g t t

6900 (in frame with Cas9) VYIMADKQKNGIKVNFKIRH 150 155 160 165 VYIMADKQKNGIKVNFKIRH EGFP VYIMADKQKNGIKVNFKIRH c g t c t a t a t c a t g g c c g a c a a g c a g a a g a a c g g c a t c a a g g t g a a c t t c a a g a t c c g c c a g c a g a t a t a g t a c c g g c t g t t c g t c t t c t t g c c g t a g t t c c a c t t g a a g t t c t a g g c g g t

6960 (in frame with Cas9) NIEDGSVQLADHYQQNTPIG 170 175 180 185 NIEDGSVQLADHYQQNTPIG EGFP NIEDGSVQLADHYQQNTPIG c a a c a t c g a g g a c g g c a g c g t g c a g c t c g c c g a c c a c t a c c a g c a g a a c a c c c c c a t c g g g t t g t a g c t c c t g c c g t c g c a c g t c g a g c g g c t g g t g a t g g t c g t c t t g t g g g g g t a g c c

7020 (in frame with Cas9) DGPVLLPDNHYLSTQSALSK 190 195 200 205 DGPVLLPDNHYLSTQSALSK EGFP DGPVLLPDNHYLSTQSALSK c g a c g g c c c c g t g c t g c t g c c c g a c a a c c a c t a c c t g a g c a c c c a g t c c g c c c t g a g c a a g c t g c c g g g g c a c g a c g a c g g g c t g t t g g t g a t g g a c t c g t g g g t c a g g c g g g a c t c g t t

7080 (in frame with Cas9) DPNEKRDHMVLLEFVTAAGI 210 215 220 225 DPNEKRDHMVLLEFVTAAGI EGFP C A T G G T C C T G C T G G A G T T C G T G EGFP-C DPNEKRDHMVLLEFVTAAGI a g a c c c c a a c g a g a a g c g c g a t c a c a t g g t c c t g c t g g a g t t c g t g a c c g c c g c c g g g a t t c t g g g g t t g c t c t t c g c g c t a g t g t a c c a g g a c g a c c t c a a g c a c t g g c g g c g g c c c t a

PacI 7140 (in frame with Cas9) TLGMDELYK* 230 235 TLGMDELYK* EGFP TLGMDELYK* *F c a c t c t c g g c a t g g a c g a g c t g t a c a a g t a a t t a a t t a a c t a g a a t g a a t c g t t t t t a a a g t g a g a g c c g t a c c t g c t c g a c a t g t t c a t t a a t t a a t t g a t c t t a c t t a g c a a a a a t t t

7200 LLDITKYYEYSEKIIYYFTI a t a a c a a a t c a a t t g t t t t a t a a t a t t c g t a c g a t t c t t t g a t t a t g t a a t a a a a t g t g a t a t t g t t t a g t t a a c a a a a t a t t a t a a g c a t g c t a a g a a a c t a a t a c a t t a t t t t a c a c t

7260 MLFIVFFIFFILETHIVFAT t c a t t a g g a a g a t t a c g a a a a a t a t a a a a a a t a t g a g t t c t g t g t g t a t a a c a a a t g c t g a g t a a t c c t t c t a a t g c t t t t t a t a t t t t t t a t a c t c a a g a c a c a c a t a t t g t t t a c g a c

7320 FAVITNTAFLGMIIQNFNNN t a a a c g c c a c a a t t g t g t t t g t t g c a a a t a a a c c c a t g a t t a t t t g a t t a a a a t t g t t g t a t t t g c g g t g t t a a c a c a a a c a a c g t t t a t t t g g g t a c t a a t a a a c t a a t t t t a a c a a c a

7380 EKNMSLLTNQRFTYQMFEMR t t t c t t t g t t c a t a g a c a a t a g t g t g t t t t g c c t a a a c g t g t a c t g c a t a a a c t c c a t g c a a a g a a a c a a g t a t c t g t t a t c a c a c a a a a c g g a t t t g c a c a t g a c g t a t t t g a g g t a c g

7440 TYLSSTALAQGVLTSEDFDE g a g t g t a t a g c g a g c t a g t g g c t a a c g c t t g c c c c a c c a a a g t a g a t t c g t c a a a a t c c t c t c a c a t a t c g c t c g a t c a c c g a t t g c g a a c g g g g t g g t t t c a t c t a a g c a g t t t t a g g a

7500 IEDGEELNLMQGDSNVELSA c a a t t t c a t c a c c c t c c t c c a a g t t t a a c a t t t g g c c g t c g g a a t t a a c t t c t a a a g a t g g t t a a a g t a g t g g g a g g a g g t t c a a a t t g t a a a c c g g c a g c c t t a a t t g a a g a t t t c t a c

7560 VYDLLHFLSEFTADDDTEVM c c a c a t a a t c t a a t a a a t g a a a t a g a g a t t c a a a c g t g g c g t c a t c g t c c g t t t c g a c c a g g t g t a t t a g a t t a t t t a c t t t a t c t c t a a g t t t g c a c c g c a g t a g c a g g c a a a g c t g g t

7620 ESFFEPMFEIIERSTTNDFS t t t c c g a a a a g a a c t c g g g c a t a a a c t c t a t g a t t t c t c t g g a c g t g g t g t t g t c g a a a c a a a g g c t t t t c t t g a g c c c g t a t t t g a g a t a c t a a a g a g a c c t g c a c c a c a a c a g c t t t g

7680 EFYATLFTRSMDDPFERPFM t c t c a a a g t a c g c a g t c a g g a a c g t g c g c g a c a t g t c g t c g g g a a a c t c g c g c g g a a a c a a g a g t t t c a t g c g t c a g t c c t t g c a c g c g c t g t a c a g c a g c c c t t t g a g c g c g c c t t t g t

7740 NNYGFPDWLALVLDALTLLI t g t t g t t g t a a c c g a a c g g g t c c c a t a g c g c c a a a a c c a a a t c t g c c a g c g t c a a t a g a a a c a a c a a c a t t g g c t t g c c c a g g g t a t c g c g g t t t t g g t t t a g a c g g t c g c a g t t a t c t t

AscI 7800 LVIGVISSAQIAIRTLPRPA t g a g c a c g a t g c c g a c a a t g g a g c t g g c t t g g a t a g c g a t t c g a g t t a a c g g c c g g g g c g a c t c g t g c t a c g g c t g t t a c c t c g a c c g a a c c t a t c g c t a a g c t c a a t t g c c g g c c c c g c

7860 GMSPPRGVVLKKASCFFVSK c g c c c a t a c t c g g t g g c c t c c c c a c c a c c a a c t t t t t t g c a c t g c a a a a a a a c a c g c t t t g c g g g t a t g a g c c a c c g g a g g g g t g g t g g t t g a a a a a a c g t g a c g t t t t t t t g t g c g a a a

7920 CAPGYM t g c a c g c g g g c c c a t a c a t a g t a c a a a c t c t a c g t t t c g t a g a c t a t t t t a c a t a a a t a g a c g t g c g c c c g g g t a t g t a t c a t g t t t g a g a t g c a a a g c a t c t g a t a a a a t g t a t t t a t c

7980 t c t a c a c c g t t g t a t a c g c t c c a a a t a c a c t a c c a c a c a t t g a a c c t t t t t g c a g t g c a a a g a t g t g g c a a c a t a t g c g a g g t t t a t g t g a t g g t g t g t a a c t t g g a a a a a c g t c a c g t t

8040 a a a a g t a c g t g t c g g c a g t c a c g t a g g c c g g c c t t a t c g g g t c g c g t c c t g t c a c g t a c g t t t t c a t g c a c a g c c g t c a g t g c a t c c g g c c g g a a t a g c c c a g c g c a g g a c a g t g c a t g c

RsrII 8100 a a t c a c a t t a t c g g a c c g g a c g a g t g t t g t c t t a t c g t g a c a g g a c g c c a g c t t c c t g t g t t a g t g t a a t a g c c t g g c c t g c t c a c a a c a g a a t a g c a c t g t c c t g c g g t c g a a g g a c a c

8160 t t g c t a a c c g c a g c c g g a c g c a a c t c c t t a t c g g a a c a g g a c g c g c c t c c a t a t c a g c c g a a c g a t t g g c g t c g g c c t g c g t t g a g g a a t a g c c t t g t c c t g c g c g g a g g t a t a g t c g g c

8220 MRVTGHEAPVPLIAPI c g c g t t a t c t c a t g c g c g t g a c c g g a c a c g a g g c g c c c g t c c c g c t t a t c g c g c c t a t a a g c g c a a t a g a g t a c g c g c a c t g g c c t g t g c t c c g c g g g c a g g g c g a a t a g c g c g g a t a t t

8280 NTARNDLVNTVEQHLLQRHN a t a c a g c c c g c a a c g a t c t g g t a a a c a c a g t t g a a c a g c a t c t g t t a c a g c g a c a c a a c a t a t g t c g g g c g t t g c t a g a c c a t t t g t g t c a a c t t g t c g t a g a c a a t g t c g c t g t g t t g t

8340 MSRSNNANAPTPSNRRRNLS t g a g c c g g t c c a a c a a c g c c a a c g c g c c c a c g c c a t c c a a c c g c c g c c g c a a c c t g t c t c a c t c g g c c a g g t t g t t g c g g t t g c g c g g g t g c g g t a g g t t g g c g g c g g c g t t g g a c a g a g

FspAI 8400 1 M 1a V 5 10 15 RSSKNVIKEFMRFKVR DsRed1 LVMVRSSKNVIKEFMRFKVR t g g t g a t g g t g c g c t c c t c c a a g a a c g t c a t c a a g g a g t t c a t g c g c t t c a a g g t g c g c a a c c a c t a c c a c g c g a g g a g g t t c t t g c a g t a g t t c c t c a a g t a c g c g a a g t t c c a c g c g t

8460 20 25 30 35 MEGTVNGHEFEIEGEGEGRP DsRed1 MEGTVNGHEFEIEGEGEGRP t g g a g g g c a c c g t g a a c g g c c a c g a g t t c g a g a t c g a g g g c g a g g g c g a g g g c c g c c c c t a c c t c c c g t g g c a c t t g c c g g t g c t c a a g c t c t a g c t c c c g c t c c c g c t c c c g g c g g g g a

8520 40 45 50 55 YEGHNTVKLKVTKGGPLPFA DsRed1 YEGHNTVKLKVTKGGPLPFA a c g a g g g c c a c a a c a c c g t g a a g c t g a a g g t g a c c a a g g g c g g c c c c c t g c c c t t c g c c t t g c t c c c g g t g t t g t g g c a c t t c g a c t t c c a c t g g t t c c c g c c g g g g g a c g g g a a g c g g a

8580 60 65 70 75 WDILSPQFQYGSKVYVKHPA DsRed1 G A C A G G G G G G T C A A G G T C A T G DsRed1-N WDILSPQFQYGSKVYVKHPA g g g a c a t c c t g t c c c c c c a g t t c c a g t a c g g c t c c a a g g t g t a c g t g a a g c a c c c c g c c g c c c t g t a g g a c a g g g g g g t c a a g g t c a t g c c g a g g t t c c a c a t g c a c t t c g t g g g g c g g c

8640 80 85 90 95 DIPDYKKLSFPEGFKWERVM DsRed1 DIPDYKKLSFPEGFKWERVM a c a t c c c c g a c t a c a a g a a g c t g t c c t t c c c c g a g g g c t t c a a g t g g g a g c g c g t g a t g a t g t a g g g g c t g a t g t t c t t c g a c a g g a a g g g g c t c c c g a a g t t c a c c c t c g c g c a c t a c t

SbfI 8700 100 105 110 115 NFEDGGVVTVTQDSSLQDGC DsRed1 NFEDGGVVTVTQDSSLQDGC a c t t c g a g g a c g g c g g c g t g g t g a c c g t g a c c c a g g a c t c c t c c c t g c a g g a c g g c t g c t t g a a g c t c c t g c c g c c g c a c c a c t g g c a c t g g g t c c t g a g g a g g g a c g t c c t g c c g a c g a

8760 120 125 130 135 FIYKVKFIGVNFPSDGPVMQ DsRed1 C C C C G T A A T G C A G A mCherry-F FIYKVKFIGVNFPSDGPVMQ t c a t c t a c a a g g t g a a g t t c a t c g g c g t g a a c t t c c c c t c c g a c g g c c c c g t a a t g c a g a a g t a g a t g t t c c a c t t c a a g t a g c c g c a c t t g a a g g g g a g g c t g c c g g g g c a t t a c g t c t

StuINcoI 8820 140 145 150 155 KKTMGWEASTERLYPRDGVL DsRed1 A G A A G A mCherry-F KKTMGWEASTERLYPRDGVL a g a a g a c c a t g g g c t g g g a g g c c t c c a c c g a g c g c c t g t a c c c c c g c g a c g g c g t g c t g a t c t t c t g g t a c c c g a c c c t c c g g a g g t g g c t c g c g g a c a t g g g g g c g c t g c c g c a c g a c t

8880 160 165 170 175 KGEIHKALKLKDGGHYLVEF DsRed1 KGEIHKALKLKDGGHYLVEF a g g g c g a g a t c c a c a a g g c c c t g a a g c t g a a g g a c g g c g g c c a c t a c c t g g t g g a g t t c a t c c c g c t c t a g g t g t t c c g g g a c t t c g a c t t c c t g c c g c c g g t g a t g g a c c a c c t c a a g t

8940 180 185 190 195 KSIYMAKKPVQLPGYYYVDS DsRed1 KSIYMAKKPVQLPGYYYVDS a g t c c a t c t a c a t g g c c a a g a a g c c c g t g c a g c t g c c c g g c t a c t a c t a c g t g g a c t c c a t c a g g t a g a t g t a c c g g t t c t t c g g g c a c g t c g a c g g g c c g a t g a t g a t g c a c c t g a g g t

9000 200 205 210 215 KLDITSHNEDYTIVEQYERT DsRed1 A G C T G G A C A T C A C C T C C C A C A A C G DsRed1-C KLDITSHNEDYTIVEQYERT a g c t g g a c a t c a c c t c c c a c a a c g a g g a c t a c a c c a t c g t g g a g c a g t a c g a g c g c a c c g t c g a c c t g t a g t g g a g g g t g t t g c t c c t g a t g t g g t a g c a c c t c g t c a t g c t c g c g t g g c

9060 220 225 EGRHHLFL* DsRed1 EGRHHLFL* a g g g c c g c c a c c a c c t g t t c c t g t a g g a c t c t a g a t c a t a a t c a g c c a t a c c a c a t t t g t t c c c g g c g g t g g t g g a c a a g g a c a t c c t g a g a t c t a g t a t t a g t c g g t a t g g t g t a a a c a

9120 a g a g g t t t t a c t t g c t t t a a a a a a c c t c c c a c a c c t c c c c c t g a a c c t g a a a c a t a a a a t t c t c c a a a a t g a a c g a a a t t t t t t g g a g g g t g t g g a g g g g g a c t t g g a c t t t g t a t t t t a

9180 SV40 poly(A) signal C G T T SV40pA-R g a a t g c a a t t g t t g t t g t t a a c t t g t t t a t t g c a g c t t a t a a t g g t t a c a a a t a a a g c a a c t t a c g t t a a c a a c a a c a a t t g a a c a a a t a a c g t c g a a t a t t a c c a a t g t t t a t t t c g t t

9240 SV40 poly(A) signal G T G G T T T G T C EBV-rev A T C G T A G T G T T T A A A G SV40pA-R t a g c a t c a c a a a t t t c a c a a a t a a a g c a t t t t t t t c a c t g c a t t c t a g t t g t g g t t t g t c a t c g t a g t g t t t a a a g t g t t t a t t t c g t a a a a a a a g t g a c g t a a g a t c a a c a c c a a a c a g

9300 SV40 poly(A) signal C A A A C T C A T C EBV-rev c a a a c t c a t c a a t g t a t c t t a a a g c t t c t a g g c c g g c c g a t c t c g g a t c t g a c a a t g t t c g t t t g a g t a g t t a c a t a g a a t t t c g a a g a t c c g g c c g g c t a g a g c c t a g a c t g t t a c a a g

9360 a g t g c a g a g a c t c g g c t a c g c c t c g t g g a c t t t g a a g t t g a c c a a c a a t g t t t a t t c t t a t c a c g t c t c t g a g c c g a t g c g g a g c a c c t g a a a c t t c a a c t g g t t g t t a c a a a t a a g a a t

9420 c c t c t a a t a g t c c t c t g t g g c a a g g t c a a g a t t c t g t t a g a a g c c a a t g a a g a a c c t g g t g g a g a t t a t c a g g a g a c a c c g t t c c a g t t c t a a g a c a a t c t t c g g t t a c t t c t t g g a c c a

9480 t g t t c a a t a a c a t t t t g t t c g t c t a a t a t t t c a c t a c c g c t t g a c g t t g g c t g c a c t t c a a c a a g t t a t t g t a a a a c a a g c a g a t t a t a a a g t g a t g g c g a a c t g c a a c c g a c g t g a a g t

9540 t g t a c c t c a t c t a t a a a c g c t t c t t c t g t a t c g c t c t g g a c g t c a t c t t c a c t t a c g t g a a c a t g g a g t a g a t a t t t g c g a a g a a g a c a t a g c g a g a c c t g c a g t a g a a g t g a a t g c a c t

9600 t c t g a t a t t t c a c t g t c a g a a t c c t c a c c a a c a a g c t c g t c a t c g c t t t g c a g a a g a g c a a g a c t a t a a a g t g a c a g t c t t a g g a g t g g t t g t t c g a g c a g t a g c g a a a c g t c t t c t c g t

9660 g a g a g g a t a t g c t c a t c g t c t a a a g a a c t a c c c a t t t t a t t a t a t a t t a g t c a c g a t a t c c t c t c c t a t a c g a g t a g c a g a t t t c t t g a t g g g t a a a a t a a t a t a t a a t c a g t g c t a t a g

9720 t a t a a c a a g a a a a t a t a t a t a t a a t a a g t t a t c a c g t a a g t a g a a c a t g a a a t a a c a a t a a t a t t g t t c t t t t a t a t a t a t a t t a t t c a a t a g t g c a t t c a t c t t g t a c t t t a t t g t t a t

9780 t a a t t a t c g t a t g a g t t a a a t c t t a a a a g t c a c g t a a a a g a t a a t c a t g c g t c a t t t t g a a t t a a t a g c a t a c t c a a t t t a g a a t t t t c a g t g c a t t t t c t a t t a g t a c g c a g t a a a a c t

9840 c t c a c g c g g t c g t t a t a g t t c a a a a t c a g t g a c a c t t a c c g c a t t g a c a a g c a c g c c t c a g a g t g c g c c a g c a a t a t c a a g t t t t a g t c a c t g t g a a t g g c g t a a c t g t t c g t g c g g a g t

9900 c g g g a g c t c c a a g c g g c g a c t g a g a t g t c c t a a a t g c a c a g c g a c g g a t t c g c g c t a t t t g c c c t c g a g g t t c g c c g c t g a c t c t a c a g g a t t t a c g t g t c g c t g c c t a a g c g c g a t a a a

9960 piggyBac left (5') inverted repeat a g a a a g a g a g a g c a a t a t t t c a a g a a t g c a t g c g t c a a t t t t a c g c a g a c t a t c t t t c t a t c t t t c t c t c t c g t t a t a a a g t t c t t a c g t a c g c a g t t a a a a t g c g t c t g a t a g a a a g a t

10,020 piggyBac left (5') inverted repeat g g g t t a a a a a a g a t t t g c g c t t t a c t c g a c c t a a a c t t t a a a c a c g t c a t a g a a t c t t c g c c c a a t t t t t t c t a a a c g c g a a a t g a g c t g g a t t t g a a a t t t g t g c a g t a t c t t a g a a g c

10,080 t t t g a c a a a a a c c a c a t t g t g g c c a a g c t g t g t g a c g c g a c g c g c g c t a a a g a a t g g c a a a a a c t g t t t t t g g t g t a a c a c c g g t t c g a c a c a c t g c g c t g c g c g c g a t t t c t t a c c g t t

EcoRIBamHIAcc65IKpnI 10,140 a c c a a g t c g c g c g a g c g t c g a c t c t a g a g g a t c c c c g g g t a c c g a g c t c g a a t t c g t a a t t g g t t c a g c g c g c t c g c a g c t g a g a t c t c c t a g g g g c c c a t g g c t c g a g c t t a a g c a t t a

10,200 -10 M13 rev lac operator lac promoter C A G T A T C G A C A A A G G A C M13 Reverse G G A C A C A C T T T A A C A A T A G G C G A M13/pUC Reverse c a t g t c a t a g c t g t t t c c t g t g t g a a a t t g t t a t c c g c t c a c a a t t c c a c a c a a c a t a c g g t a c a g t a t c g a c a a a g g a c a c a c t t t a a c a a t a g g c g a g t g t t a a g g t g t g t t g t a t g c

10,260 -35 lac promoter CAP binding site a g c c g g a a g c a t a a a g t g t a a a g c c t g g g g t g c c t a a t g a g t g a g c t a a c t c a c a t t a a t t c g g c c t t c g t a t t t c a c a t t t c g g a c c c c a c g g a t t a c t c a c t c g a t t g a g t g t a a t t a

10,320 t g c g t t g c g c t c a c t g c c c g c t t t c c a g t c g g g a a a c c t g t c g t g c c a g c t g c a t t a a t g a c g c a a c g c g a g t g a c g g g c g a a a g g t c a g c c c t t t g g a c a g c a c g g t c g a c g t a a t t a c

10,380 G A G C G A L4440 a a t c g g c c a a c g c g c g g g g a g a g g c g g t t t g c g t a t t g g g c g c t c t t c c g c t t c c t c g c t t t a g c c g g t t g c g c g c c c c t c t c c g c c a a a c g c a t a a c c c g c g a g a a g g c g a a g g a g c g a

10,440 G T G A C T G A G C G A L4440 c a c t g a c t c g c t g c g c t c g g t c g t t c g g c t g c g g c g a g c g g t a t c a g c t c a c t c a a a g g c g t g a c t g a g c g a c g c g a g c c a g c a a g c c g a c g c c g c t c g c c a t a g t c g a g t g a g t t t c c g

10,500 g g t a a t a c g g t t a t c c a c a g a a t c a g g g g a t a a c g c a g g a a a g a a c a t g t g a g c a a a a g g c c a t t a t g c c a a t a g g t g t c t t a g t c c c c t a t t g c g t c c t t t c t t g t a c a c t c g t t t t c c

10,560 ori c c a g c a a a a g g c c a g g a a c c g t a a a a a g g c c g c g t t g c t g g c g t t t t t c c a t a g g c t c c g g g t c g t t t t c c g g t c c t t g g c a t t t t t c c g g c g c a a c g a c c g c a a a a a g g t a t c c g a g g c

10,620 ori c c c c c c t g a c g a g c a t c a c a a a a a t c g a c g c t c a a g t c a g a g g t g g c g a a a c c c g a c a g g g g g g g g a c t g c t c g t a g t g t t t t t a g c t g c g a g t t c a g t c t c c a c c g c t t t g g g c t g t c c

10,680 ori T T T C T A T G G T C C G C A A A G G G pBR322ori-F a c t a t a a a g a t a c c a g g c g t t t c c c c c t g g a a g c t c c c t c g t g c g c t c t c c t g t t c c g a c t g a t a t t t c t a t g g t c c g c a a a g g g g g a c c t t c g a g g g a g c a c g c g a g a g g a c a a g g c t g

10,740 ori c c t g c c g c t t a c c g g a t a c c t g t c c g c c t t t c t c c c t t c g g g a a g c g t g g c g c t t t c t c a g g a c g g c g a a t g g c c t a t g g a c a g g c g g a a a g a g g g a a g c c c t t c g c a c c g c g a a a g a g t

10,800 ori t a g c t c a c g c t g t a g g t a t c t c a g t t c g g t g t a g g t c g t t c g c t c c a a g c t g g g c t g t g t a t c g a g t g c g a c a t c c a t a g a g t c a a g c c a c a t c c a g c a a g c g a g g t t c g a c c c g a c a c a

10,860 ori g c a c g a a c c c c c c g t t c a g c c c g a c c g c t g c g c c t t a t c c g g t a a c t a t c g t c t t g a g t c c g t g c t t g g g g g g c a a g t c g g g c t g g c g a c g c g g a a t a g g c c a t t g a t a g c a g a a c t c a g

10,920 ori c a a c c c g g t a a g a c a c g a c t t a t c g c c a c t g g c a g c a g c c a c t g g t a a c a g g a t t a g c a g g t t g g g c c a t t c t g t g c t g a a t a g c g g t g a c c g t c g t c g g t g a c c a t t g t c c t a a t c g t c

10,980 ori a g c g a g g t a t g t a g g c g g t g c t a c a g a g t t c t t g a a g t g g t g g c c t a a c t a c g g c t a c a c t c g c t c c a t a c a t c c g c c a c g a t g t c t c a a g a a c t t c a c c a c c g g a t t g a t g c c g a t g t g

11,040 ori t a g a a g a a c a g t a t t t g g t a t c t g c g c t c t g c t g a a g c c a g t t a c c t t c g g a a a a a g a g t a t c t t c t t g t c a t a a a c c a t a g a c g c g a g a c g a c t t c g g t c a a t g g a a g c c t t t t t c t c a

11,100 ori t g g t a g c t c t t g a t c c g g c a a a c a a a c c a c c g c t g g t a g c g g t g g t t t t t t t g t t t g c a a a c c a t c g a g a a c t a g g c c g t t t g t t t g g t g g c g a c c a t c g c c a c c a a a a a a a c a a a c g t t

11,160 ori g c a g c a g a t t a c g c g c a g a a a a a a a g g a t c t c a a g a a g a t c c t t t g a t c t t t t c t a c g g g c g t c g t c t a a t g c g c g t c t t t t t t t c c t a g a g t t c t t c t a g g a a a c t a g a a a a g a t g c c c

11,220 g t c t g a c g c t c a g t g g a a c g a a a a c t c a c g t t a a g g g a t t t t g g t c a t g a g a t t a t c a a a c a g a c t g c g a g t c a c c t t g c t t t t g a g t g c a a t t c c c t a a a a c c a g t a c t c t a a t a g t t t

11,280 a a g g a t c t t c a c c t a g a t c c t t t t a a a t t a a a a a t g a a g t t t t a a a t c a a t c t a a a g t a t t t c c t a g a a g t g g a t c t a g g a a a a t t t a a t t t t t a c t t c a a a a t t t a g t t a g a t t t c a t a

11,340 285 280 *WHKILSAGIEA AmpR *WHKILSAGIEA a t a t g a g t a a a c t t g g t c t g a c a g t t a c c a a t g c t t a a t c a g t g a g g c a c c t a t c t c a g c t a t a c t c a t t t g a a c c a g a c t g t c a a t g g t t a c g a a t t a g t c a c t c c g t g g a t a g a g t c g

11,400 275 270 265 260 IQRNREDMTAQSGTTYIVVI AmpR IQRNREDMTAQSGTTYIVVI g a t c t g t c t a t t t c g t t c a t c c a t a g t t g c c t g a c t c c c c g t c g t g t a g a t a a c t a c g a t c t a g a c a g a t a a a g c a a g t a g g t a t c a a c g g a c t g a g g g g c a g c a c a t c t a t t g a t g c t a

11,460 255 250 245 240 RSPKGDPGLAAIIGRSGREG AmpR RSPKGDPGLAAIIGRSGREG a c g g g a g g g c t t a c c a t c t g g c c c c a g t g c t g c a a t g a t a c c g c g a g a c c c a c g c t c a c c t g c c c t c c c g a a t g g t a g a c c g g g g t c a c g a c g t t a c t a t g g c g c t c t g g g t g c g a g t g g

11,520 235 230 225 220 AGSKDAIFWGAPLASRLLPG AmpR AGSKDAIFWGAPLASRLLPG g g c t c c a g a t t t a t c a g c a a t a a a c c a g c c a g c c g g a a g g g c c g a g c g c a g a a g t g g t c c c c g a g g t c t a a a t a g t c g t t a t t t g g t c g g t c g g c c t t c c c g g c t c g c g t c t t c a c c a g g

11,580 215 210 205 200 AVKDAEMWDILQQRSALTLL AmpR AVKDAEMWDILQQRSALTLL t g c a a c t t t a t c c g c c t c c a t c c a g t c t a t t a a t t g t t g c c g g g a a g c t a g a g t a a g t a g a c g t t g a a a t a g g c g g a g g t a g g t c a g a t a a t t a a c a a c g g c c c t t c g a t c t c a t t c a t c

11,640 195 190 185 180 EGTLLKRLTTAMAVPMTTDR AmpR EGTLLKRLTTAMAVPMTTDR t t c g c c a g t t a a t a g t t t g c g c a a c g t t g t t g c c a t t g c t a c a g g c a t c g t g g t g t c a c g a a g c g g t c a a t t a t c a a a c g c g t t g c a a c a a c g g t a a c g a t g t c c g t a g c a c c a c a g t g c

11,700 175 170 165 160 EDNPIAENLEPEWRDLRTVH AmpR EDNPIAENLEPEWRDLRTVH c t c g t c g t t t g g t a t g g c t t c a t t c a g c t c c g g t t c c c a a c g a t c a a g g c g a g t t a c a t g g a g c a g c a a a c c a t a c c g a a g t a a g t c g a g g c c a a g g g t t g c t a g t t c c g c t c a a t g t a c

11,760 155 150 145 140 DGMNHLFATLEKPGGITTLL AmpR DGMNHLFATLEKPGGITTLL a t c c c c c a t g t t g t g c a a a a a a g c g g t t a g c t c c t t c g g t c c t c c g a t c g t t g t c a g a a g t a g g g g g t a c a a c a c g t t t t t t c g c c a a t c g a g g a a g c c a g g a g g c t a g c a a c a g t c t t c

11,820 135 130 125 120 LNAATNDSMTIAASCLERVT AmpR LNAATNDSMTIAASCLERVT t a a g t t g g c c g c a g t g t t a t c a c t c a t g g t t a t g g c a g c a c t g c a t a a t t c t c t t a c t g t a t t c a a c c g g c g t c a c a a t a g t g a g t a c c a a t a c c g t c g t g a c g t a t t a a g a g a a t g a c a

11,880 115 110 105 100 MGDTLHKETVPSYEVLDNQS AmpR MGDTLHKETVPSYEVLDNQS c a t g c c a t c c g t a a g a t g c t t t t c t g t g a c t g g t g a g t a c t c a a c c a a g t c a t t c t g a g a g t a c g g t a g g c a t t c t a c g a a a a g a c a c t g a c c a c t c a t g a g t t g g t t c a g t a a g a c t c t

11,940 95 90 85 80 YHIRRGLQEQGADIRSLVAG AmpR A T A A T A C C G C G C C Amp-R YHIRRGLQEQGADIRSLVAG a t a g t g t a t g c g g c g a c c g a g t t g c t c t t g c c c g g c g t c a a t a c g g g a t a a t a c c g c g c c t a t c a c a t a c g c c g c t g g c t c a a c g a g a a c g g g c c g c a g t t a t g c c c t a t t a t g g c g c g g

12,000 75 70 65 60 CLLVKFTSMMPFREEPRFSE AmpR A C A T A G C Amp-R CLLVKFTSMMPFREEPRFSE a c a t a g c a g a a c t t t a a a a g t g c t c a t c a t t g g a a a a c g t t c t t c g g g g c g a a a a c t c t c t g t a t c g t c t t g a a a t t t t c a c g a g t a g t a a c c t t t t g c a a g a a g c c c c g c t t t t g a g a g

12,060 55 50 45 40 LIKGSNLDLEIYGVRAGLQD AmpR LIKGSNLDLEIYGVRAGLQD a a g g a t c t t a c c g c t g t t g a g a t c c a g t t c g a t g t a a c c c a c t c g t g c a c c c a a c t g a t c t t c c t a g a a t g g c g a c a a c t c t a g g t c a a g c t a c a t t g g g t g a g c a c g t g g g t t g a c t a g

12,120 35 30 25 EADKVKVLTEPH 20 signal sequence AFVPLCFA AmpR EADKVKVLTEPHAFVPLCFA t t c a g c a t c t t t t a c t t t c a c c a g c g t t t c t g g g t g a g c a a a a a c a g g a a g g c a a a a t g c a a g t c g t a g a a a a t g a a a g t g g t c g c a a a g a c c c a c t c g t t t t t g t c c t t c c g t t t t a c g

12,180 15 10 5 1 signal sequence AFFPILAVRFHQISM AmpR AmpR promoter AFFPILAVRFHQISM c g c a a a a a a g g g a a t a a g g g c g a c a c g g a a a t g t t g a a t a c t c a t a c t c t t c c t t t t t c a g c g t t t t t t c c c t t a t t c c c g c t g t g c c t t t a c a a c t t a t g a g t a t g a g a a g g a a a a a g t

12,240 AmpR promoter a t a t t a t t g a a g c a t t t a t c a g g g t t a t t g t c t c a t g a g c g g a t a c a t a t t t g a a t g t a t t a t a a t a a c t t c g t a a a t a g t c c c a a t a a c a g a g t a c t c g c c t a t g t a t a a a c t t a c a t a

12,300 AmpR promoter t t a g a a a a a t a a a c a a a t a g g g g t t c c g c g c a c a t t t c c c c g a a a a g t g c c a c c t g a c g t a a t c t t t t t a t t t g t t t a t c c c c a a g g c g c g t g t a a a g g g g c t t t t c a c g g t g g a c t g c a

12,360 A A T A G G C G T A T C A C G A G G C pBRforEco c t a a g a a a c c a t t a t t a t c a t g a c a t t a a c c t a t a a a a a t a g g c g t a t c a c g a g g c c c t t g a t t c t t t g g t a a t a a t a g t a c t g t a a t t g g a t a t t t t t a t c c g c a t a g t g c t c c g g g a a

12,420 G G A G A C T G T G T A C G T C G A G G G C C pGEX 3' t c g t c t c g c g c g t t t c g g t g a t g a c g g t g a a a a c c t c t g a c a c a t g c a g c t c c c g g a g a c a g c a g a g c g c g c a a a g c c a c t a c t g c c a c t t t t g g a g a c t g t g t a c g t c g a g g g c c t c t g

12,480 g g t c a c a g c t t g t c t g t a a g c g g a t g c c g g g a g c a g a c a a g c c c g t c a g g g c g c g t c a g c c c a g t g t c g a a c a g a c a t t c g c c t a c g g c c c t c g t c t g t t c g g g c a g t c c c g c g c a g t c g

12,540 C G G C A T C A G A G C A G A T T G T A pRS-marker g g g t g t t g g c g g g t g t c g g g g c t g g c t t a a c t a t g c g g c a t c a g a g c a g a t t g t a c t g a g c c c a c a a c c g c c c a c a g c c c c g a c c g a a t t g a t a c g c c g t a g t c t c g t c t a a c a t g a c t c

NdeI 12,600 a g t g c a c c a t a t g c g g t g t g a a a t a c c g c a c a g a t g c g t a a g g a g a a a a t a c c g c a t c a g t c a c g t g g t a t a c g c c a c a c t t t a t g g c g t g t c t a c g c a t t c c t c t t t t a t g g c g t a g t c

12,660 *AACSNPLAIPAPRK g c g c c a t t c g c c a t t c a g g c t g c g c a a c t g t t g g g a a g g g c g a t c g g t g c g g g c c t c t t c c g c g g t a a g c g g t a a g t c c g a c g c g t t g a c a a c c c t t c c c g c t a g c c a c g c c c g g a g a a g

12,720 AIVGAPSLPIHQLAILNPLA g c t a t t a c g c c a g c t g g c g a a a g g g g g a t g t g c t g c a a g g c g a t t a a g t t g g g t a a c g c c c g a t a a t g c g g t c g a c c g c t t t c c c c c t a c a c g a c g t t c c g c t a a t t c a a c c c a t t g c g g

12,780 M13 fwd C C C A G T C A C G A C G T T G T A A A A C G M13/pUC Forward T G T A A A A C G A C G G C C A G T M13 Forward LTKGTVVNYFSPWHWAKNLF a g g g t t t t c c c a g t c a c g a c g t t g t a a a a c g a c g g c c a g t g c c a a g c t t t g t t t a a a a a t t c c c a a a a g g g t c a g t g c t g c a a c a t t t t g c t g c c g g t c a c g g t t c g a a a c a a a t t t t t a

12,840 IVFNHDWLIFHPLILYNITD a t a a c a a a a t t g t g a t c c c a c a a a a t g a a g t g g g g c a a a a t c a a a t a a t t a a t a g t g t c c t a t t g t t t t a a c a c t a g g g t g t t t t a c t t c a c c c c g t t t t a g t t t a t t a a t t a t c a c a g g

12,900 TFKNTKLKKLFVNSPLDTVI g t a a a c t t g t t g g t c t t c a a c t t t t t g a g g a a c a c g t t g g a c g g c a a a t c c g t g a c t a t a c a t t t g a a c a a c c a g a a g t t g a a a a a c t c c t t g t g c a a c c t g c c g t t t a g g c a c t g a t a t

12,960 VCTSKIIKALVDPQTNKASA a c a c a a g t t g a t t t a a t a a t t t t a g c c a a c a c g t c g g g c t g c g t g t t t t t t g c c g a c g c g t g t g t t c a a c t a a a t t a t t a a a a t c g g t t g t g c a g c c c g a c g c a c a a a a a a c g g c t g c g c

13,020 DTYVNILQDILSNFYNLKQN t c t g t g t a c a c g t t g a t t a a c t g g t c g a t t a a a c t g t t g a a a t a a t t t a a t t t t t g g t t c a g a c a c a t g t g c a a c t a a t t g a c c a g c t a a t t t g a c a a c t t t a t t a a a t t a a a a a c c a a g

13,080 KKLDTIFNKLIVKFEENTFF t t c t t t a a a t c t g t g a t g a a a t t t t t t a a a a t a a c t t t a a a t t c t t c a t t g g t a a a a a a t a a g a a a t t t a g a c a c t a c t t t a a a a a a t t t t a t t g a a a t t t a a g a a g t a a c c a t t t t t t a

13,140 AVNQLKHPDLILDFETPTKD g c c a c g t t t t g c a a c t t g t g a g g g t c t a a t a t g a g g t c a a a c t c a g t a g g a g t t t t a t c c c g g t g c a a a a c g t t g a a c a c t c c c a g a t t a t a c t c c a g t t t g a g t c a t c c t c a a a a t a g g

3ʹ 5ʹ 13,159 LFSFMIV a a a a a a g a a a a c a t g a t t a t t t t t t c t t t t g t a c t a a t

Restriction Enzymes

Instructions: By default, all cutters are shown. Filter on number of cut sites or search by enzyme name.

Filter

Features

Primers

BLAST

BLAST (Basic Local Alignment Search Tool) finds regions of similarity between biological sequences. Click on the buttons below to submit a BLAST search to NCBI. The results will appear in a new window. See your recent BLAST results on NCBI's website.

  • Nucleotide-Nucleotide BLAST (BLASTN)

  • Translated Nucleotide-Protein BLAST (BLASTX)

  • Sequence alignment using BLAST (BLAST2)

Sequence Analyzer Guide

Map

Displays a graphical map based on nucleotide sequence data labeled with restriction enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site)

To select a portion of sequence, click one location on the plasmid and then a second location to display the sequence between the two locations.

Sequence

Displays both strands of base paired nucleotide sequences with annotated enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site).

To select a portion of sequence, click one location on the sequence and then a second location to display the sequence between the two locations.

Enzymes

List of restriction enzymes that can cut a given nucleotide sequence. Table lists enzyme name and the sequence location of the cut.

Features

List of common features detected in a given nucleotide sequence. Table lists feature name, location, size, color used to indicate its position on the map, and direction (if relevant).

Primers

List of commonly used primers detected in a given nucleotide sequence. Table lists primer name, sequence, length, binding site location, and direction.

BLAST

Use Basic Local Alignment Search Tool (BLAST) via the NCBI website to determine similarity between a given sequence and nucleotide (BLASTN) or protein (BLASTX) sequence databases. Additionally, align a custom nucleotide sequence against a given sequence using BLAST2.

File Downloads

GenBank

File contains the nucleotide sequence and annotated features in GenBank flat file format. Open the file with a text editor or plasmid mapping software to view the sequence.

SnapGene

File contains the nucleotide sequence and enhanced annotations from SnapGene Server. Open the file with SnapGene software or the free Viewer to view the plasmid map, sequence, and perform additional sequence analysis.