Addgene pYPQ269A2 Sequencing Result - Sequence Analyzer Skip to main content
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Sequence Analyzer: pYPQ269A2 Sequencing Result


Map View

M13 rev T7 (9229 .. 9248) T7 promoter attR5 BstZ17I (9098) MauBI (9016) NsiI (8912) BsaBI (8855) EagI - NotI (8777) KpnI (8721) Acc65I (8717) SV40 NLS SpeI (8417) SspI (8280) SacI (8229) Eco53kI (8227) SV40 NLS BmgBI (7924) T2A MfeI (7885) SV40 NLS PshAI (6725) MscI (6587) CsiI - SexAI * (6002) KflI (5805) SmaI (5723) TspMI - XmaI (5721) PflFI - Tth111I (5366) AbsI (5276) BmtI (4890) M13 Reverse (9253 .. 9269) BsaAI (1113) pBR322ori-F (1731 .. 1750) L4440 (1984 .. 2001) rrnB T2 terminator AclI (2256) M13/pUC Forward (2414 .. 2436) M13 Forward (2428 .. 2445) M13 fwd PspOMI (2455) ApaI (2459) PsiI (2530) BsrGI (2543) NcoI (3312) SV40 NLS SbfI (3678) BstXI (3697) SgrDI (4046) NheI (4886) pYPQ269A2 9342 bp

Sequence View

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

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

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

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

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

360 1 M SmR WSSNDVTQQGSRPKTKLNIM t g g a g c a g c a a c g a t g t t a c g c a g c a g g g c a g t c g c c c t a a a a c a a a g t t a a a c a t t a t g a c c t c g t c g t t g c t a c a a t g c g t c g t c c c g t c a g c g g g a t t t t g t t t c a a t t t g t a a t a c

420 5 10 15 20 REAVIAEVSTQLSEVVGVIE SmR REAVIAEVSTQLSEVVGVIE a g g g a a g c g g t g a t c g c c g a a g t a t c g a c t c a a c t a t c a g a g g t a g t t g g c g t c a t c g a g t c c c t t c g c c a c t a g c g g c t t c a t a g c t g a g t t g a t a g t c t c c a t c a a c c g c a g t a g c t c

480 25 30 35 40 RHLEPTLLAVHLYGSAVDGG SmR RHLEPTLLAVHLYGSAVDGG c g c c a t c t c g a a c c g a c g t t g c t g g c c g t a c a t t t g t a c g g c t c c g c a g t g g a t g g c g g c g c g g t a g a g c t t g g c t g c a a c g a c c g g c a t g t a a a c a t g c c g a g g c g t c a c c t a c c g c c g

540 45 50 55 60 LKPHSDIDLLVTVTVRLDET SmR LKPHSDIDLLVTVTVRLDET c t g a a g c c a c a c a g t g a t a t t g a t t t g c t g g t t a c g g t g a c c g t a a g g c t t g a t g a a a c a g a c t t c g g t g t g t c a c t a t a a c t a a a c g a c c a a t g c c a c t g g c a t t c c g a a c t a c t t t g t

600 65 70 75 80 TRRALINDLLETSASPGESE SmR TRRALINDLLETSASPGESE a c g c g g c g a g c t t t g a t c a a c g a c c t t t t g g a a a c t t c g g c t t c c c c t g g a g a g a g c g a g t g c g c c g c t c g a a a c t a g t t g c t g g a a a a c c t t t g a a g c c g a a g g g g a c c t c t c t c g c t c

660 85 90 95 100 ILRAVEVTIVVHDDIIPWRY SmR ILRAVEVTIVVHDDIIPWRY a t t c t c c g c g c t g t a g a a g t c a c c a t t g t t g t g c a c g a c g a c a t c a t t c c g t g g c g t t a t t a a g a g g c g c g a c a t c t t c a g t g g t a a c a a c a c g t g c t g c t g t a g t a a g g c a c c g c a a t a

720 105 110 115 120 PAKRELQFGEWQRNDILAGI SmR PAKRELQFGEWQRNDILAGI c c a g c t a a g c g c g a a c t g c a a t t t g g a g a a t g g c a g c g c a a t g a c a t t c t t g c a g g t a t c g g t c g a t t c g c g c t t g a c g t t a a a c c t c t t a c c g t c g c g t t a c t g t a a g a a c g t c c a t a g

780 125 130 135 140 FEPATIDIDLAILLTKAREH SmR FEPATIDIDLAILLTKAREH t t c g a g c c a g c c a c g a t c g a c a t t g a t c t g g c t a t c t t g c t g a c a a a a g c a a g a g a a c a t a a g c t c g g t c g g t g c t a g c t g t a a c t a g a c c g a t a g a a c g a c t g t t t t c g t t c t c t t g t a

840 145 150 155 160 SVALVGPAAEELFDPVPEQD SmR SVALVGPAAEELFDPVPEQD a g c g t t g c c t t g g t a g g t c c a g c g g c g g a g g a a c t c t t t g a t c c g g t t c c t g a a c a g g a t t c g c a a c g g a a c c a t c c a g g t c g c c g c c t c c t t g a g a a a c t a g g c c a a g g a c t t g t c c t a

900 165 170 175 180 LFEALNETLTLWNSPPDWAG SmR LFEALNETLTLWNSPPDWAG c t a t t t g a g g c g c t a a a t g a a a c c t t a a c g c t a t g g a a c t c g c c g c c c g a c t g g g c t g g c g a t a a a c t c c g c g a t t t a c t t t g g a a t t g c g a t a c c t t g a g c g g c g g g c t g a c c c g a c c g

960 185 190 195 200 DERNVVLTLSRIWYSAVTGK SmR DERNVVLTLSRIWYSAVTGK g a t g a g c g a a a t g t a g t g c t t a c g t t g t c c c g c a t t t g g t a c a g c g c a g t a a c c g g c a a a c t a c t c g c t t t a c a t c a c g a a t g c a a c a g g g c g t a a a c c a t g t c g c g t c a t t g g c c g t t t

1020 205 210 215 220 IAPKDVAADWAMERLPAQYQ SmR IAPKDVAADWAMERLPAQYQ a t c g c g c c g a a g g a t g t c g c t g c c g a c t g g g c a a t g g a g c g c c t g c c g g c c c a g t a t c a g t a g c g c g g c t t c c t a c a g c g a c g g c t g a c c c g t t a c c t c g c g g a c g g c c g g g t c a t a g t c

1080 225 230 235 240 PVILEARQAYLGQEEDRLAS SmR PVILEARQAYLGQEEDRLAS c c c g t c a t a c t t g a a g c t a g a c a g g c t t a t c t t g g a c a a g a a g a a g a t c g c t t g g c c t c g g g g c a g t a t g a a c t t c g a t c t g t c c g a a t a g a a c c t g t t c t t c t t c t a g c g a a c c g g a g c

BsaAI 1140 245 250 255 260 RADQLEEFVHYVKGEITKVV SmR RADQLEEFVHYVKGEITKVV c g c g c a g a t c a g t t g g a a g a a t t t g t c c a c t a c g t g a a a g g c g a g a t c a c c a a g g t a g t c g c g c g t c t a g t c a a c c t t c t t a a a c a g g t g a t g c a c t t t c c g c t c t a g t g g t t c c a t c a g

1200 GK* SmR GK* g g c a a a t a a c c c t c g a g c c a c c c a t g a c c a a a a t c c c t t a a c g t g a g t t a c g c g t c g t t c c c g t t t a t t g g g a g c t c g g t g g g t a c t g g t t t t a g g g a a t t g c a c t c a a t g c g c a g c a a g

1260 ori c a c t g a g c g t c a g a c c c c g t a g a a a a g a t c a a a g g a t c t t c t t g a g a t c c t t t t t t t c t g g t g a c t c g c a g t c t g g g g c a t c t t t t c t a g t t t c c t a g a a g a a c t c t a g g a a a a a a a g a c

1320 ori c g c g t a a t c t g c t g c t t g c a a a c a a a a a a a c c a c c g c t a c c a g c g g t g g t t t g t t t g c c g g c g c a t t a g a c g a c g a a c g t t t g t t t t t t t g g t g g c g a t g g t c g c c a c c a a a c a a a c g g c

1380 ori g a t c a a g a g c t a c c a a c t c t t t t t c c g a a g g t a a c t g g c t t c a g c a g a g c g c a g a t a c c a c t a g t t c t c g a t g g t t g a g a a a a a g g c t t c c a t t g a c c g a a g t c g t c t c g c g t c t a t g g t

1440 ori a a t a c t g t t c t t c t a g t g t a g c c g t a g t t a g g c c a c c a c t t c a a g a a c t c t g t a g c a c c g t t a t g a c a a g a a g a t c a c a t c g g c a t c a a t c c g g t g g t g a a g t t c t t g a g a c a t c g t g g c

1500 ori c c t a c a t a c c t c g c t c t g c t a a t c c t g t t a c c a g t g g c t g c t g c c a g t g g c g a t a a g t c g g g a t g t a t g g a g c g a g a c g a t t a g g a c a a t g g t c a c c g a c g a c g g t c a c c g c t a t t c a g c

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

1620 ori a c g g g g g g t t c g t g c a c a c a g c c c a g c t t g g a g c g a a c g a c c t a c a c c g a a c t g a g a t a c t g c c c c c c a a g c a c g t g t g t c g g g t c g a a c c t c g c t t g c t g g a t g t g g c t t g a c t c t a t g

1680 ori c t a c a g c g t g a g c t a t g a g a a a g c g c c a c g c t t c c c g a a g g g a g a a a g g c g g a c a g g t a t g a t g t c g c a c t c g a t a c t c t t t c g c g g t g c g a a g g g c t t c c c t c t t t c c g c c t g t c c a t a

1740 ori G G G A A A C G C C pBR322ori-F c c g g t a a g c g g c a g g g t c g g a a c a g g a g a g c g c a c g a g g g a g c t t c c a g g g g g a a a c g c c g g c c a t t c g c c g t c c c a g c c t t g t c c t c t c g c g t g c t c c c t c g a a g g t c c c c c t t t g c g g

1800 ori T G G T A T C T T T pBR322ori-F t g g t a t c t t t a t a g t c c t g t c g g g t t t c g c c a c c t c t g a c t t g a g c g t c g a t t t t t g t g a a c c a t a g a a a t a t c a g g a c a g c c c a a a g c g g t g g a g a c t g a a c t c g c a g c t a a a a a c a c t

1860 ori t g c t c g t c a g g g g g g c g g a g c c t a t g g a a a a a c g c c a g c a a c g c g g c c t t t t t a c g g t t c a c g a g c a g t c c c c c c g c c t c g g a t a c c t t t t t g c g g t c g t t g c g c c g g a a a a a t g c c a a g

1920 c t g g c c t t t t g c t g g c c t t t t g c t c a c a t g t t c t t t c c t g c g t t a t c c c c t g a t t c t g t g g a c c g g a a a a c g a c c g g a a a a c g a g t g t a c a a g a a a g g a c g c a a t a g g g g a c t a a g a c a c

1980 g a t a a c c g t a t t a c c g c c t t t g a g t g a g c t g a t a c c g c t c g c c g c a g c c g a a c g a c c g a g c t a t t g g c a t a a t g g c g g a a a c t c a c t c g a c t a t g g c g a g c g g c g t c g g c t t g c t g g c t c

2040 A G C G A G T C A G T G A G C G A G L4440 c g c a g c g a g t c a g t g a g c g a g g a a g c g g a a g a g c g c c c a a t a c g c a a a c c g c c t c t c c c c g c g t c g c t c a g t c a c t c g c t c c t t c g c c t t c t c g c g g g t t a t g c g t t t g g c g g a g a g g g g

2100 g c g c g t t g g c c g a t t c a t t a a t g c a g c t g g c a c g a c a g g t t t c c c g a c t g g a a a g c g g g c c g c g c a a c c g g c t a a g t a a t t a c g t c g a c c g t g c t g t c c a a a g g g c t g a c c t t t c g c c c g

2160 rrnB T2 terminator a g t g a g c g c a a c g c a a t t a a t a c g c g t a c c g c g a g c c a g g a a g a g t t t g t a g a a a c g c a a t c a c t c g c g t t g c g t t a a t t a t g c g c a t g g c g c t c g g t c c t t c t c a a a c a t c t t t g c g t t

2220 rrnB T2 terminator a a a g g c c a t c c g t c a g g a t g g c c t t c t g c t t a g t t t g a t g c c t g g c a g t t t a t g g c g g g c t t t c c g g t a g g c a g t c c t a c c g g a a g a c g a a t c a a a c t a c g g a c c g t c a a a t a c c g c c c g

AclI 2280 rrnB T1 terminator g t c c t g c c c g c c a c c c t c c g g g c c g t t g c t t c a c a a c g t t c a a a t c c g c t c c c g g c g g a t c a g g a c g g g c g g t g g g a g g c c c g g c a a c g a a g t g t t g c a a g t t t a g g c g a g g g c c g c c t a

2340 rrnB T1 terminator t t g t c c t a c t c a g g a g a g c g t t c a c c g a c a a a c a a c a g a t a a a a c g a a a g g c c c a g t c t t a a c a g g a t g a g t c c t c t c g c a a g t g g c t g t t t g t t g t c t a t t t t g c t t t c c g g g t c a g a a

2400 rrnB T1 terminator c c g a c t g a g c c t t t c g t t t t a t t t g a t g c c t g g c a g t t c c c t a c t c t c g c g t t a a c g c t t g g c t g a c t c g g a a a g c a a a a t a a a c t a c g g a c c g t c a a g g g a t g a g a g c g c a a t t g c g a a

PspOMIApaI 2460 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 g c a t g g a t 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 c t t a a g c t c g g g c c c c g t a c c t a 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 g a a t t c g a g c c c g g g

2520 attL1 c a a a t a a t g a t t t t a t t t t g a c t g a t a g t g a c c t g t t c g t t g c a a c a a a t t g a t g a g c a a g t t t a t t a c t a a a a t a a a a c t g a c t a t c a c t g g a c a a g c a a c g t t g t t t a a c t a c t c g t t

BsrGIPsiI 2580 1 MS attL1 APOBEC-1 MS t g c t t t t t t a t a a t g c c a a c t t t g t a c a a a a a a g c a g g c t c c g a a t t c g c c c t t c a t g a g a c g a a a a a a t a t t a c g g t t g a a a c a t g t t t t t t c g t c c g a g g c t t a a g c g g g a a g t a c t c

2640 5 10 15 20 SETGPVAVDPTLRRRIEPHE APOBEC-1 SETGPVAVDPTLRRRIEPHE t t c t g a a a c a g g t c c a g t g g c t g t c g a t c c g a c t c t g a g g a g g c g c a t c g a a c c a c a c g a a a g a c t t t g t c c a g g t c a c c g a c a g c t a g g c t g a g a c t c c t c c g c g t a g c t t g g t g t g c t

2700 25 30 35 40 FEVFFDPRELRKETCLLYEI APOBEC-1 FEVFFDPRELRKETCLLYEI a t t t g a g g t t t t t t t t g a t c c a a g a g a g c t t c g c a a g g a a a c t t g t c t c c t g t a t g a g a t t a a a c t c c a a a a a a a a c t a g g t t c t c t c g a a g c g t t c c t t t g a a c a g a g g a c a t a c t c t a

2760 45 50 55 60 NWGGRHSIWRHTSQNTNKHV APOBEC-1 NWGGRHSIWRHTSQNTNKHV a a a t t g g g g a g g a a g a c a t a g t a t c t g g a g a c a c a c a a g c c a g a a c a c c a a t a a g c a t g t t t t a a c c c c t c c t t c t g t a t c a t a g a c c t c t g t g t g t t c g g t c t t g t g g t t a t t c g t a c a

2820 65 70 75 80 EVNFIEKFTTERYFCPNTRC APOBEC-1 EVNFIEKFTTERYFCPNTRC c g a a g t t a a c t t c a t c g a a a a a t t t a c g a c c g a a c g c t a c t t t t g t c c t a a t a c a c g c t g g c t t c a a t t g a a g t a g c t t t t t a a a t g c t g g c t t g c g a t g a a a a c a g g a t t a t g t g c g a c

2880 85 90 95 100 SITWFLSWSPCGECSRAITE APOBEC-1 SITWFLSWSPCGECSRAITE c a g t a t c a c t t g g t t c c t t t c g t g g t c a c c t t g c g g t g a a t g t t c c a g a g c a a t t a c g g a g t c a t a g t g a a c c a a g g a a a g c a c c a g t g g a a c g c c a c t t a c a a g g t c t c g t t a a t g c c t

2940 105 110 115 120 FLSRYPHVTLFIYIARLYHH APOBEC-1 FLSRYPHVTLFIYIARLYHH g t t t c t c t c g a g a t a t c c a c a c g t t a c a c t t t t c a t t t a c a t a g c t c g g c t t t a t c a t c a c a a a g a g a g c t c t a t a g g t g t g c a a t g t g a a a a g t a a a t g t a t c g a g c c g a a a t a g t a g t

3000 125 130 135 140 ADPRNRQGLRDLISSGVTIQ APOBEC-1 ADPRNRQGLRDLISSGVTIQ c g c a g a t c c g a g a a a t c g c c a g g g g c t g c g c g a t c t g a t t t c g t c c g g t g t c a c c a t t c a g c g t c t a g g c t c t t t a g c g g t c c c c g a c g c g c t a g a c t a a a g c a g g c c a c a g t g g t a a g t

3060 145 150 155 160 IMTEQESGYCWRNFVNYSPS APOBEC-1 IMTEQESGYCWRNFVNYSPS a a t t a t g a c t g a g c a g g a a t c t g g a t a c t g c t g g c g c a a c t t c g t g a a c t a c a g t c c g t c t t a a t a c t g a c t c g t c c t t a g a c c t a t g a c g a c c g c g t t g a a g c a c t t g a t g t c a g g c a g

3120 165 170 175 180 NEAHWPRYPHLWVRLYVLEL APOBEC-1 NEAHWPRYPHLWVRLYVLEL g a a t g a a g c g c a c t g g c c g a g a t a t c c c c a t c t c t g g g t g c g c t t g t a t g t t t t g g a g c t c t t a c t t c g c g t g a c c g g c t c t a t a g g g g t a g a g a c c c a c g c g a a c a t a c a a a a c c t c g a

3180 185 190 195 200 YCIILGLPPCLNILRRKQPQ APOBEC-1 YCIILGLPPCLNILRRKQPQ t t a t t g t a t c a t c t t g g g a t t g c c a c c t t g t c t t a a t a t a t t g c g c c g g a a a c a a c c g c a a a t a a c a t a g t a g a a c c c t a a c g g t g g a a c a g a a t t a t a t a a c g c g g c c t t t g t t g g c g t

3240 205 210 215 220 LTFFTIALQSCHYQRLPPHI APOBEC-1 LTFFTIALQSCHYQRLPPHI g c t c a c t t t t t t c a c c a t t g c t c t g c a g t c t t g t c a c t a c c a a c g c c t c c c c c c g c a c a t c g a g t g a a a a a a g t g g t a a c g a g a c g t c a g a a c a g t g a t g g t t g c g g a g g g g g g c g t g t a

3300 225 LWATGLK (in frame with APOBEC-1) SGSETPGTSESAT APOBEC-1 LWATGLKSGSETPGTSESAT c c t g t g g g c c a c a g g c c t t a a a t c a g g t t c t g a g a c c c c t g g c a c a a g t g a g t c a g c a a c g g a c a c c c g g t g t c c g g a a t t t a g t c c a a g a c t c t g g g g a c c g t g t t c a c t c a g t c g t t g

NcoI 3360 (in frame with APOBEC-1) PESTM 1 5 10 15 DYKDHDGDYKDHDID 3xFLAG PESTMDYKDHDGDYKDHDID a c c c g a g t c c a c c a t g g a t t a c a a g g a c c a c g a c g g g g a t t a c a a g g a c c a c g a c a t t g a t g g g c t c a g g t g g t a c c t a a t g t t c c t g g t g c t g c c c c t a a t g t t c c t g g t g c t g t a a c t

3420 20 YKDDDDK MA 130 PKKKRKV GIHG 3xFLAG SV40 NLS YKDDDDKMAPKKKRKVGIHG t t a c a a g g a t g a t g a t g a c a a g a t g g c t c c g a a g a a g a a g a g g a a g g t t g g c a t c c a c g g a a t g t t c c t a c t a c t a c t g t t c t a c c g a g g c t t c t t c t t c t c c t t c c a a c c g t a g g t g c c

3480 VPAA 5 DKKYSIGL 10 A 15 IGTNSVG Cas9(D10A) VPAADKKYSIGLAIGTNSVG g g t g c c a g c t g c t g a c a a g a a g t a c t c g a t c g g c c t c g c t a t t g g g a c t a a c t c t g t t g g c c a c g g t c g a c g a c t g t t c t t c a t g a g c t a g c c g g a g c g a t a a c c c t g a t t g a g a c a a c c

3540 20 25 30 35 WAVITDEYKVPSKKFKVLGN Cas9(D10A) WAVITDEYKVPSKKFKVLGN 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 t c a a a g a a g t t c a a g g t c c t g g g c a a 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 a g t t t c t t c a a g t t c c a g g a c c c g t t

3600 40 45 50 55 TDRHSIKKNLIGALLFDSGE Cas9(D10A) TDRHSIKKNLIGALLFDSGE c a c c g a t c g g c a t t c c a t c a a g a a g a a t c t c a t t g g c g c t c t c c t g t t c g a c a g c g g c g a g t g g c t a g c c g t a a g g t a g t t c t t c t t a g a g t a a c c g c g a g a g g a c a a g c t g t c g c c g c t

3660 60 65 70 75 TAEATRLKRTARRRYTRRKN Cas9(D10A) TAEATRLKRTARRRYTRRKN g a c g g c t g a g g c t a c g c g g c t c a a g c g c a c c g c c c g c a g g c g g t a c a c g c g c a g g a a g a a c t g c c g a c t c c g a t g c g c c g a g t t c g c g t g g c g g g c g t c c g c c a t g t g c g c g t c c t t c t t

BstXISbfI 3720 80 85 90 95 RICYLQEIFSNEMAKVDDSF Cas9(D10A) RICYLQEIFSNEMAKVDDSF t c g c a t c t g c t a c c t g c a g g a g a t t t t c t c c a a c g a g a t g g c g a a g g t t g a c g a t t c t t t a g c g t a g a c g a t g g a c g t c c t c t a a a a g a g g t t g c t c t a c c g c t t c c a a c t g c t a a g a a a

3780 100 105 110 115 FHRLEESFLVEEDKKHERHP Cas9(D10A) FHRLEESFLVEEDKKHERHP c t t c c a c a g g c t g g a g g a g t c a t t c c t c g t g g a g g a g g a t a a g a a g c a c g a g c g g c a t c c g a a g g t g t c c g a c c t c c t c a g t a a g g a g c a c c t c c t c c t a t t c t t c g t g c t c g c c g t a g g

3840 120 125 130 135 IFGNIVDEVAYHEKYPTIYH Cas9(D10A) IFGNIVDEVAYHEKYPTIYH a a t c t t c g g c a a c a t t g t c g a c g a g g t t g c c t a c c a c g a g a a g t a c c c t a c g a t c t a c c a t t a g a a g c c g t t g t a a c a g c t g c t c c a a c g g a t g g t g c t c t t c a t g g g a t g c t a g a t g g t

3900 140 145 150 155 LRKKLVDSTDKADLRLIYLA Cas9(D10A) LRKKLVDSTDKADLRLIYLA t c t g c g g a a g a a g c t c g t g g a c t c c a c a g a t a a g g c g g a c c t c c g c c t g a t c t a c c t c g c a g a c g c c t t c t t c g a g c a c c t g a g g t g t c t a t t c c g c c t g g a g g c g g a c t a g a t g g a g c g

3960 160 165 170 175 LAHMIKFRGHFLIEGDLNPD Cas9(D10A) LAHMIKFRGHFLIEGDLNPD t c t g g c c c a c a t g a t t a a g t t c a g g g g c c a t t t c c t g a t c g a g g g g g a t c t c a a c c c g g a a g a c c g g g t g t a c t a a t t c a a g t c c c c g g t a a a g g a c t a g c t c c c c c t a g a g t t g g g c c t

4020 180 185 190 195 NSDVDKLFIQLVQTYNQLFE Cas9(D10A) NSDVDKLFIQLVQTYNQLFE c a a t a g c g a t g t t g a c a a g c t g t t c a t c c a g c t c g t g c a g a c g t a c a a c c a g c t c t t c g a g t t a t c g c t a c a a c t g t t c g a c a a g t a g g t c g a g c a c g t c t g c a t g t t g g t c g a g a a g c t

SgrDI 4080 200 205 210 215 ENPINASGVDAKAILSARLS Cas9(D10A) ENPINASGVDAKAILSARLS g g a g a a c c c c a t t a a t g c g t c a g g c g t c g a c g c g a a g g c t a t c c t g t c c g c t a g g c t c t c c c t c t t g g g g t a a t t a c g c a g t c c g c a g c t g c g c t t c c g a t a g g a c a g g c g a t c c g a g a g

4140 220 225 230 235 KSRRLENLIAQLPGEKKNGL Cas9(D10A) KSRRLENLIAQLPGEKKNGL g a a g t c t c g g c g c c t c g a g a a c c t g a t c g c c c a g c t g c c g g g c g a g a a g a a g a a c g g c c t c t t c a g a g c c g c g g a g c t c t t g g a c t a g c g g g t c g a c g g c c c g c t c t t c t t c t t g c c g g a

4200 240 245 250 255 FGNLIALSLGLTPNFKSNFD Cas9(D10A) FGNLIALSLGLTPNFKSNFD g t t c g g g a a t c t c a t t g c g c t c a g c c t g g g g c t c a c g c c c a a c t t c a a g t c g a a t t t c g a c a a g c c c t t a g a g t a a c g c g a g t c g g a c c c c g a g t g c g g g t t g a a g t t c a g c t t a a a g c t

4260 260 265 270 275 LAEDAKLQLSKDTYDDDLDN Cas9(D10A) LAEDAKLQLSKDTYDDDLDN t c t c g c t g a g g a c g c c a a g c t g c a g c t c t c c a a g g a c a c a t a c g a c g a t g a c c t g g a t a a a g a g c g a c t c c t g c g g t t c g a c g t c g a g a g g t t c c t g t g t a t g c t g c t a c t g g a c c t a t t

4320 280 285 290 295 LLAQIGDQYADLFLAAKNLS Cas9(D10A) LLAQIGDQYADLFLAAKNLS c c t c c t g g c c c a g a t c g g c g a t c a g t a c g c g g a c c t g t t c c t c g c t g c c a a g a a t c t g t c g g a g g a c c g g g t c t a g c c g c t a g t c a t g c g c c t g g a c a a g g a g c g a c g g t t c t t a g a c a g

4380 300 305 310 315 DAILLSDILRVNTEITKAPL Cas9(D10A) DAILLSDILRVNTEITKAPL g g a c g c c a t c c t c c t g t c t g a t a t t c t c a g g g t g a a c a c c g a g a t t a c g a a g g c t c c g c t c c t g c g g t a g g a g g a c a g a c t a t a a g a g t c c c a c t t g t g g c t c t a a t g c t t c c g a g g c g a

4440 320 325 330 335 SASMIKRYDEHHQDLTLLKA Cas9(D10A) SASMIKRYDEHHQDLTLLKA c t c a g c c t c c a t g a t c a a g c g c t a c g a c g a g c a c c a t c a g g a t c t g a c c c t c c t g a a g g c g a g t c g g a g g t a c t a g t t c g c g a t g c t g c t c g t g g t a g t c c t a g a c t g g g a g g a c t t c c g

4500 340 345 350 355 LVRQQLPEKYKEIFFDQSKN Cas9(D10A) LVRQQLPEKYKEIFFDQSKN g c t g g t c a g g c a g c a g c t c c c c g a g a a g t a c a a g g a g a t c t t c t t c g a t c a g t c g a a g a a c g a c c a g t c c g t c g t c g a g g g g c t c t t c a t g t t c c t c t a g a a g a a g c t a g t c a g c t t c t t

4560 360 365 370 375 GYAGYIDGGASQEEFYKFIK Cas9(D10A) GYAGYIDGGASQEEFYKFIK c g g c t a c g c t g g g t a c a t t g a c g g c g g g g c c t c t c a g g a g g a g t t c t a c a a g t t c a t c a a g c c g a t g c g a c c c a t g t a a c t g c c g c c c c g g a g a g t c c t c c t c a a g a t g t t c a a g t a g t t

4620 380 385 390 395 PILEKMDGTEELLVKLNRED Cas9(D10A) PILEKMDGTEELLVKLNRED g c c g a t t c t g g a g a a g a t g g a c g g c a c g g a g g a g c t g c t g g t g a a g c t c a a t c g c g a g g a c g g c t a a g a c c t c t t c t a c c t g c c g t g c c t c c t c g a c g a c c a c t t c g a g t t a g c g c t c c t

4680 400 405 410 415 LLRKQRTFDNGSIPHQIHLG Cas9(D10A) LLRKQRTFDNGSIPHQIHLG c c t c c t g a g g a a g c a g c g g a c a t t c g a t a a c g g c a g c a t c c c a c a c c a g a t t c a t c t c g g g g a g g a c t c c t t c g t c g c c t g t a a g c t a t t g c c g t c g t a g g g t g t g g t c t a a g t a g a g c c

4740 420 425 430 435 ELHAILRRQEDFYPFLKDNR Cas9(D10A) ELHAILRRQEDFYPFLKDNR g g a g c t g c a c g c t a t c c t g a g g a g g c a g g a g g a c t t c t a c c c t t t c c t c a a g g a t a a c c g c c t c g a c g t g c g a t a g g a c t c c t c c g t c c t c c t g a a g a t g g g a a a g g a g t t c c t a t t g g c

4800 440 445 450 455 EKIEKILTFRIPYYVGPLAR Cas9(D10A) EKIEKILTFRIPYYVGPLAR c g a g a a g a t c g a g a a g a t t c t g a c t t t c a g g a t c c c g t a c t a c g t c g g c c c a c t c g c t a g g c t c t t c t a g c t c t t c t a a g a c t g a a a g t c c t a g g g c a t g a t g c a g c c g g g t g a g c g a t c

4860 460 465 470 475 GNSRFAWMTRKSEETITPWN Cas9(D10A) GNSRFAWMTRKSEETITPWN *LLRDRRPV g g g c a a c t c c c g c t t c g c t t g g a t g a c c c g c a a g t c a g a g g a g a c g a t c a c g c c g t g g a a c c c g t t g a g g g c g a a g c g a a c c t a c t g g g c g t t c a g t c t c c t c t g c t a g t g c g g c a c c t t

BmtINheI 4920 480 485 490 495 FEEVVDKGASAQSFIERMTN Cas9(D10A) FEEVVDKGASAQSFIERMTN ELLHDVLASASLREDLPHRI c t t c g a g g a g g t g g t c g a c a a g g g c g c t a g c g c t c a g t c g t t c a t c g a g a g g a t g a c g a a g a a g c t c c t c c a c c a g c t g t t c c c g c g a t c g c g a g t c a g c a a g t a g c t c t c c t a c t g c t t

4980 500 505 510 515 FDKNLPNEKVLPKHSLLYEY Cas9(D10A) FDKNLPNEKVLPKHSLLYEY EVLVQWILLHERLVREQVLV t t t c g a c a a g a a c c t g c c a a a t g a g a a g g t g c t c c c t a a g c a c t c g c t c c t g t a c g a g t a a a a g c t g t t c t t g g a c g g t t t a c t c t t c c a c g a g g g a t t c g t g a g c g a g g a c a t g c t c a t

5040 520 525 530 535 FTVYNELTKVKYVTEGMRKP Cas9(D10A) FTVYNELTKVKYVTEGMRKP ECDVVLQSLHLIHGLAHPLR c t t c a c a g t c t a c a a c g a g c t g a c t a a g g t g a a g t a t g t g a c c g a g g g c a t g a g g a a g c c g a a g t g t c a g a t g t t g c t c g a c t g a t t c c a c t t c a t a c a c t g g c t c c c g t a c t c c t t c g g

5100 540 545 550 555 AFLSGEQKKAIVDLLFKTNR Cas9(D10A) AFLSGEQKKAIVDLLFKTNR SEQRPLLLLGDHVEQELGVP g g c t t t c c t g t c t g g g g a g c a g a a g a a g g c c a t c g t g g a c c t c c t g t t c a a g a c c a a c c g c c g a a a g g a c a g a c c c c t c g t c t t c t t c c g g t a g c a c c t g g a g g a c a a g t t c t g g t t g g c

5160 560 565 570 575 KVTVKQLKEDYFKKIECFDS Cas9(D10A) KVTVKQLKEDYFKKIECFDS LDRNLLELLVVELLNLAEIR g a a g g t c a c g g t t a a g c a g c t c a a g g a g g a c t a c t t c a a g a a g a t t g a g t g c t t c g a t t c c t t c c a g t g c c a a t t c g t c g a g t t c c t c c t g a t g a a g t t c t t c t a a c t c a c g a a g c t a a g

5220 580 585 590 595 VEISGVEDRFNASLGTYHDL Cas9(D10A) VEISGVEDRFNASLGTYHDL DLDRANLVAEVGGQPGVVIE g g t c g a g a t c t c t g g c g t t g a g g a c c g c t t c a a c g c c t c c c t g g g g a c c t a c c a c g a t c t c c a g c t c t a g a g a c c g c a a c t c c t g g c g a a g t t g c g g a g g g a c c c c t g g a t g g t g c t a g a

AbsI 5280 600 605 610 615 LKIIKDKDFLDNEENEDILE Cas9(D10A) LKIIKDKDFLDNEENEDILE QLDNLILVEQVVLLILIDEL c c t g a a g a t c a t t a a g g a t a a g g a c t t c c t g g a c a a c g a g g a g a a t g a g g a t a t c c t c g a g g a c t t c t a g t a a t t c c t a t t c c t g a a g g a c c t g t t g c t c c t c t t a c t c c t a t a g g a g c t

5340 620 625 630 635 DIVLTLTLFEDREMIEERLK Cas9(D10A) DIVLTLTLFEDREMIEERLK VNHQCESQELVPLHDLLAQL g g a c a t t g t g c t g a c a c t c a c t c t g t t c g a g g a c c g g g a g a t g a t c g a g g a g c g c c t g a a c c t g t a a c a c g a c t g t g a g t g a g a c a a g c t c c t g g c c c t c t a c t a g c t c c t c g c g g a c t t

Tth111IPflFI 5400 640 645 650 655 TYAHLFDDKVMKQLKRRRYT Cas9(D10A) TYAHLFDDKVMKQLKRRRYT SVGMEEIVLDHLLELPPPVG g a c t t a c g c c c a t c t c t t c g a t g a c a a g g t c a t g a a g c a g c t c a a g a g g a g g a g g t a c a c c t g a a t g c g g g t a g a g a a g c t a c t g t t c c a g t a c t t c g t c g a g t t c t c c t c c t c c a t g t g

5460 660 665 670 675 GWGRLSRKLINGIRDKQSGK Cas9(D10A) GWGRLSRKLINGIRDKQSGK APPPQAPLEDVANPVLLGPL c g g c t g g g g g a g g c t g a g c a g g a a g c t c a t c a a c g g c a t t c g g g a c a a g c a g t c c g g g a a g c c g a c c c c c t c c g a c t c g t c c t t c g a g t a g t t g c c g t a a g c c c t g t t c g t c a g g c c c t t

5520 680 685 690 695 TILDFLKSDGFANRNFMQLI Cas9(D10A) TILDFLKSDGFANRNFMQLI RDEVEQLAIAERVAIEHLQN g a c g a t c c t c g a c t t c c t g a a g a g c g a t g g c t t c g c g a a c c g c a a t t t c a t g c a g c t g a t c t g c t a g g a g c t g a a g g a c t t c t c g c t a c c g a a g c g c t t g g c g t t a a a g t a c g t c g a c t a

5580 700 705 710 715 HDDSLTFKEDIQKAQVSGQG Cas9(D10A) HDDSLTFKEDIQKAQVSGQG VIVAECELLIDLLSLHAALP t c a c g a t g a c a g c c t c a c a t t c a a g g a g g a t a t c c a g a a g g c t c a g g t g a g c g g c c a g g g a g t g c t a c t g t c g g a g t g t a a g t t c c t c c t a t a g g t c t t c c g a g t c c a c t c g c c g g t c c c

5640 720 725 730 735 DSLHEHIANLAGSPAIKKGI Cas9(D10A) DSLHEHIANLAGSPAIKKGI VRQVLM g g a c t c g c t g c a c g a g c a t a t c g c g a a c c t c g c t g g c t c g c c a g c t a t c a a g a a g g g g a t c c t g a g c g a c g t g c t c g t a t a g c g c t t g g a g c g a c c g a g c g g t c g a t a g t t c t t c c c c t a

5700 740 745 750 755 LQTVKVVDELVKVMGRHKPE Cas9(D10A) LQTVKVVDELVKVMGRHKPE t c t g c a g a c c g t g a a g g t t g t g g a c g a g c t g g t g a a g g t c a t g g g c a g g c a c a a g c c t g a a g a c g t c t g g c a c t t c c a a c a c c t g c t c g a c c a c t t c c a g t a c c c g t c c g t g t t c g g a c t

TspMIXmaISmaI 5760 760 765 770 775 NIVIEMARENQTTQKGQKNS Cas9(D10A) NIVIEMARENQTTQKGQKNS g a a c a t c g t c a t t g a g a t g g c c c g g g a g a a t c a g a c c a c g c a g a a g g g c c a g a a g a a c t c c t t g t a g c a g t a a c t c t a c c g g g c c c t c t t a g t c t g g t g c g t c t t c c c g g t c t t c t t g a g

KflI 5820 780 785 790 795 RERMKRIEEGIKELGSQILK Cas9(D10A) RERMKRIEEGIKELGSQILK a c g c g a g a g g a t g a a g a g g a t c g a g g a g g g c a t t a a g g a g c t g g g g t c c c a g a t c c t c a a t g c g c t c t c c t a c t t c t c c t a g c t c c t c c c g t a a t t c c t c g a c c c c a g g g t c t a g g a g t t

5880 800 805 810 815 EHPVENTQLQNEKLYLYYLQ Cas9(D10A) EHPVENTQLQNEKLYLYYLQ g g a g c a c c c g g t g g a g a a c a c g c a g c t g c a g a a t g a g a a g c t c t a c c t g t a c t a c c t c c a c c t c g t g g g c c a c c t c t t g t g c g t c g a c g t c t t a c t c t t c g a g a t g g a c a t g a t g g a g g t

5940 820 825 830 835 NGRDMYVDQELDINRLSDYD Cas9(D10A) NGRDMYVDQELDINRLSDYD g a a t g g c c g c g a t a t g t a t g t g g a c c a g g a g c t g g a t a t t a a c a g g c t c a g c g a t t a c g a c t t a c c g g c g c t a t a c a t a c a c c t g g t c c t c g a c c t a t a a t t g t c c g a g t c g c t a a t g c t

6000 840 845 850 855 VDHIVPQSFLKDDSIDNKVL Cas9(D10A) VDHIVPQSFLKDDSIDNKVL c g t c g a t c a t a t c g t t c c a c a g t c a t t c c t g a a g g a t g a c t c c a t t g a c a a c a a g g t c c t g c a g c t a g t a t a g c a a g g t g t c a g t a a g g a c t t c c t a c t g a g g t a a c t g t t g t t c c a g g a

SexAI*CsiI 6060 860 865 870 875 TRSDKNRGKSDNVPSEEVVK Cas9(D10A) TRSDKNRGKSDNVPSEEVVK c a c c a g g t c g g a c a a g a a c c g g g g c a a g t c t g a t a a t g t t c c t t c a g a g g a g g t c g t t a a g t g g t c c a g c c t g t t c t t g g c c c c g t t c a g a c t a t t a c a a g g a a g t c t c c t c c a g c a a t t

6120 880 885 890 895 KMKNYWRQLLNAKLITQRKF Cas9(D10A) KMKNYWRQLLNAKLITQRKF g a a g a t g a a g a a c t a c t g g c g c c a g c t c c t g a a t g c c a a g c t g a t c a c g c a g c g g a a g t t c t t c t a c t t c t t g a t g a c c g c g g t c g a g g a c t t a c g g t t c g a c t a g t g c g t c g c c t t c a a

6180 900 905 910 915 DNLTKAERGGLSELDKAGFI Cas9(D10A) DNLTKAERGGLSELDKAGFI c g a t a a c c t c a c a a a g g c t g a g a g g g g c g g g c t c t c t g a g c t g g a c a a g g c g g g c t t c a t g c t a t t g g a g t g t t t c c g a c t c t c c c c g c c c g a g a g a c t c g a c c t g t t c c g c c c g a a g t a

6240 920 925 930 935 KRQLVETRQITKHVAQILDS Cas9(D10A) KRQLVETRQITKHVAQILDS c a a g a g g c a g c t g g t c g a g a c a c g g c a g a t c a c t a a g c a c g t t g c g c a g a t t c t c g a c t c g t t c t c c g t c g a c c a g c t c t g t g c c g t c t a g t g a t t c g t g c a a c g c g t c t a a g a g c t g a g

6300 940 945 950 955 RMNTKYDENDKLIREVKVIT Cas9(D10A) RMNTKYDENDKLIREVKVIT a c g g a t g a a c a c t a a g t a c g a t g a g a a t g a c a a g c t g a t c c g c g a g g t g a a g g t c a t c a c t g c c t a c t t g t g a t t c a t g c t a c t c t t a c t g t t c g a c t a g g c g c t c c a c t t c c a g t a g t g

6360 960 965 970 975 LKSKLVSDFRKDFQFYKVRE Cas9(D10A) LKSKLVSDFRKDFQFYKVRE c c t g a a g t c a a a g c t c g t c t c c g a c t t c a g g a a g g a t t t c c a g t t c t a c a a g g t t c g g g a g g a c t t c a g t t t c g a g c a g a g g c t g a a g t c c t t c c t a a a g g t c a a g a t g t t c c a a g c c c t

6420 980 985 990 995 INNYHHAHDAYLNAVVGTAL Cas9(D10A) INNYHHAHDAYLNAVVGTAL g a t c a a c a a t t a c c a c c a t g c c c a t g a c g c g t a c c t g a a c g c g g t g g t c g g c a c a g c t c t c t a g t t g t t a a t g g t g g t a c g g g t a c t g c g c a t g g a c t t g c g c c a c c a g c c g t g t c g a g a

6480 1000 1005 1010 1015 IKKYPKLESEFVYGDYKVYD Cas9(D10A) IKKYPKLESEFVYGDYKVYD g a t c a a g a a g t a c c c a a a g c t c g a g a g c g a g t t c g t g t a c g g g g a c t a c a a g g t t t a c g a c t a g t t c t t c a t g g g t t t c g a g c t c t c g c t c a a g c a c a t g c c c c t g a t g t t c c a a a t g c t

6540 1020 1025 1030 1035 VRKMIAKSEQEIGKATAKYF Cas9(D10A) VRKMIAKSEQEIGKATAKYF t g t g a g g a a g a t g a t c g c c a a g t c g g a g c a g g a g a t t g g c a a g g c t a c c g c c a a g t a c t t a c a c t c c t t c t a c t a g c g g t t c a g c c t c g t c c t c t a a c c g t t c c g a t g g c g g t t c a t g a a

MscI 6600 1040 1045 1050 1055 FYSNIMNFFKTEITLANGEI Cas9(D10A) FYSNIMNFFKTEITLANGEI c t t c t a c t c t a a c a t t a t g a a t t t c t t c a a g a c a g a g a t c a c t c t g g c c a a t g g c g a g a t g a a g a t g a g a t t g t a a t a c t t a a a g a a g t t c t g t c t c t a g t g a g a c c g g t t a c c g c t c t a

6660 1060 1065 1070 1075 RKRPLIETNGETGEIVWDKG Cas9(D10A) RKRPLIETNGETGEIVWDKG c c g g a a g c g c c c c c t c a t c g a g a c g a a c g g c g a g a c g g g g g a g a t c g t g t g g g a c a a g g g g g c c t t c g c g g g g g a g t a g c t c t g c t t g c c g c t c t g c c c c c t c t a g c a c a c c c t g t t c c c

6720 1080 1085 1090 1095 RDFATVRKVLSMPQVNIVKK Cas9(D10A) RDFATVRKVLSMPQVNIVKK c a g g g a t t t c g c g a c c g t c a g g a a g g t t c t c t c c a t g c c a c a a g t g a a t a t c g t c a a g a a g t c c c t a a a g c g c t g g c a g t c c t t c c a a g a g a g g t a c g g t g t t c a c t t a t a g c a g t t c t t

PshAI 6780 1100 1105 1110 1115 TEVQTGGFSKESILPKRNSD Cas9(D10A) TEVQTGGFSKESILPKRNSD g a c a g a g g t c c a g a c t g g c g g g t t c t c t a a g g a g t c a a t t c t g c c t a a g c g g a a c a g c g a c t g t c t c c a g g t c t g a c c g c c c a a g a g a t t c c t c a g t t a a g a c g g a t t c g c c t t g t c g c t

6840 1120 1125 1130 1135 KLIARKKDWDPKKYGGFDSP Cas9(D10A) KLIARKKDWDPKKYGGFDSP c a a g c t c a t c g c c c g c a a g a a g g a c t g g g a t c c g a a g a a g t a c g g c g g g t t c g a c a g c c c g t t c g a g t a g c g g g c g t t c t t c c t g a c c c t a g g c t t c t t c a t g c c g c c c a a g c t g t c g g g

6900 1140 1145 1150 1155 TVAYSVLVVAKVEKGKSKKL Cas9(D10A) TVAYSVLVVAKVEKGKSKKL c a c t g t g g c c t a c t c g g t c c t g g t t g t g g c g a a g g t t g a g a a g g g c a a g t c c a a g a a g c t g t g a c a c c g g a t g a g c c a g g a c c a a c a c c g c t t c c a a c t c t t c c c g t t c a g g t t c t t c g a

6960 1160 1165 1170 1175 KSVKELLGITIMERSSFEKN Cas9(D10A) KSVKELLGITIMERSSFEKN c a a g a g c g t g a a g g a g c t g c t g g g g a t c a c g a t t a t g g a g c g c t c c a g c t t c g a g a a g a a g t t c t c g c a c t t c c t c g a c g a c c c c t a g t g c t a a t a c c t c g c g a g g t c g a a g c t c t t c t t

7020 1180 1185 1190 1195 PIDFLEAKGYKEVKKDLIIK Cas9(D10A) PIDFLEAKGYKEVKKDLIIK c c c g a t c g a t t t c c t g g a g g c g a a g g g c t a c a a g g a g g t g a a g a a g g a c c t g a t c a t t a a g g g c t a g c t a a a g g a c c t c c g c t t c c c g a t g t t c c t c c a c t t c t t c c t g g a c t a g t a a t t

7080 1200 1205 1210 1215 LPKYSLFELENGRKRMLASA Cas9(D10A) LPKYSLFELENGRKRMLASA g c t c c c c a a g t a c t c a c t c t t c g a g c t g g a g a a c g g c a g g a a g c g g a t g c t g g c t t c c g c c g a g g g g t t c a t g a g t g a g a a g c t c g a c c t c t t g c c g t c c t t c g c c t a c g a c c g a a g g c g

7140 1220 1225 1230 1235 GELQKGNELALPSKYVNFLY Cas9(D10A) GELQKGNELALPSKYVNFLY t g g c g a g c t g c a g a a g g g g a a c g a g c t g g c t c t g c c g t c c a a g t a t g t g a a c t t c c t c t a a c c g c t c g a c g t c t t c c c c t t g c t c g a c c g a g a c g g c a g g t t c a t a c a c t t g a a g g a g a t

7200 1240 1245 1250 1255 LASHYEKLKGSPEDNEQKQL Cas9(D10A) LASHYEKLKGSPEDNEQKQL c c t g g c c t c c c a c t a c g a g a a g c t c a a g g g c a g c c c c g a g g a c a a c g a g c a g a a g c a g c t g g a c c g g a g g g t g a t g c t c t t c g a g t t c c c g t c g g g g c t c c t g t t g c t c g t c t t c g t c g a

7260 1260 1265 1270 1275 FVEQHKHYLDEIIEQISEFS Cas9(D10A) FVEQHKHYLDEIIEQISEFS g t t c g t c g a g c a g c a c a a g c a t t a c c t c g a c g a g a t c a t t g a g c a g a t t t c c g a g t t c t c c a a g c a g c t c g t c g t g t t c g t a a t g g a g c t g c t c t a g t a a c t c g t c t a a a g g c t c a a g a g

7320 1280 1285 1290 1295 KRVILADANLDKVLSAYNKH Cas9(D10A) KRVILADANLDKVLSAYNKH c a a g c g c g t g a t c c t g g c c g a c g c g a a t c t g g a t a a g g t c c t c t c c g c g t a c a a c a a g c a g t t c g c g c a c t a g g a c c g g c t g c g c t t a g a c c t a t t c c a g g a g a g g c g c a t g t t g t t c g t

7380 1300 1305 1310 1315 RDKPIREQAENIIHLFTLTN Cas9(D10A) RDKPIREQAENIIHLFTLTN c c g c g a c a a g c c a a t c a g g g a g c a g g c t g a g a a t a t c a t t c a t c t c t t c a c c c t g a c g a a g g c g c t g t t c g g t t a g t c c c t c g t c c g a c t c t t a t a g t a a g t a g a g a a g t g g g a c t g c t t

7440 1320 1325 1330 1335 LGAPAAFKYFDTTIDRKRYT Cas9(D10A) LGAPAAFKYFDTTIDRKRYT c c t c g g c g c c c c t g c t g c t t t c a a g t a c t t c g a c a c a a c t a t c g a t c g c a a g a g g t a c a c g g a g c c g c g g g g a c g a c g a a a g t t c a t g a a g c t g t g t t g a t a g c t a g c g t t c t c c a t g t g

7500 1340 1345 1350 1355 STKEVLDATLIHQSITGLYE Cas9(D10A) Shine-Dalgarno sequence STKEVLDATLIHQSITGLYE a a g c a c t a a g g a g g t c c t g g a c g c g a c c c t c a t c c a c c a g t c g a t t a c c g g c c t c t a c g a t t c g t g a t t c c t c c a g g a c c t g c g c t g g g a g t a g g t g g t c a g c t a a t g g c c g g a g a t g c t

7560 1360 1365 TRIDLSQLGGD 1 5 KRPAATKKA Cas9(D10A) nucleoplasmin NLS TRIDLSQLGGDKRPAATKKA g a c g c g c a t c g a c c t g t c t c a g c t c g g g g g c g a c a a g c g g c c a g c g g c g a c g a a g a a g g c c t g c g c g t a g c t g g a c a g a g t c g a g c c c c c g c t g t t c g c c g g t c g c c g c t g c t t c t t c c g

7620 10 15 GQAKKKK (in frame with nucleoplasmin NLS) GDGSGGS 5 TNLSDI nucleoplasmin NLS UGI GQAKKKKGDGSGGSTNLSDI g g g g c a g g c g a a g a a g a a g a a g g g a g a c g g a t c a g g c g g g t c a a c a a a c c t c a g t g a c a t c c c c g t c c g c t t c t t c t t c t t c c c t c t g c c t a g t c c g c c c a g t t g t t t g g a g t c a c t g t a

7680 10 15 20 25 IEKETGKQLVIQESILMLPE UGI IEKETGKQLVIQESILMLPE c a t a g a g a a g g a a a c t g g t a a g c a a c t g g t t a t t c a a g a g a g c a t t c t g a t g c t c c c t g a g t a t c t c t t c c t t t g a c c a t t c g t t g a c c a a t a a g t t c t c t c g t a a g a c t a c g a g g g a c t

7740 30 35 40 45 EVEEVIGNKPESDILVHTAY UGI EVEEVIGNKPESDILVHTAY g g a a g t c g a g g a a g t t a t a g g a a a c a a g c c t g a g a g c g a t a t a c t c g t g c a t a c g g c g t a c c t t c a g c t c c t t c a a t a t c c t t t g t t c g g a c t c t c g c t a t a t g a g c a c g t a t g c c g c a t

7800 50 55 60 65 DESTDENVMLLTSDAPEYKP UGI DESTDENVMLLTSDAPEYKP t g a c g a g a g c a c g g a t g a a a a t g t g a t g c t c t t g a c c a g c g a t g c g c c a g a a t a t a a a c c a c t g c t c t c g t g c c t a c t t t t a c a c t a c g a g a a c t g g t c g c t a c g c g g t c t t a t a t t t g g

7860 70 75 80 WALVIQDSNGENKIKML SGG UGI WALVIQDSNGENKIKMLSGG c t g g g c a c t g g t t a t t c a g g a c t c t a a c g g g g a a a a t a a a a t c a a g a t g c t c t c a g g t g g g a c c c g t g a c c a a t a a g t c c t g a g a t t g c c c c t t t t a t t t t a g t t c t a c g a g a g t c c a c c

MfeI 7920 S 130 PKKKRKV NW 140 145 EGRGSLLTCG SV40 NLS T2A SPKKKRKVNWEGRGSLLTCG c t c c c c a a a g a a g a a a c g c a a g g t c a a t t g g g a g g g c a g a g g a a g t c t t c t a a c a t g c g g g a g g g g t t t c t t c t t t g c g t t c c a g t t a a c c c t c c c g t c t c c t t c a g a a g a t t g t a c g c c

BmgBI 7980 150 155 DVEENPGP MA 130 PKKKRKV GIH T2A SV40 NLS DVEENPGPMAPKKKRKVGIH 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 c t c c t a a g a a g a a g c g g a a g g t t g g t a t t c a 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 g a g g a t t c t t c t t c g c c t t c c a a c c a t a a g t

8040 (in frame with SV40 NLS) GVPAAM 1 5 10 ASNFTQFVLVDNGG MS2-N55K GVPAAMASNFTQFVLVDNGG c g g g g t g c c t g c g g c t a t g g c g t c a a a t t t c a c g c a g t t t g t t t t g g t t g a t a a c g g c g g g c c c c a c g g a c g c c g a t a c c g c a g t t t a a a g t g c g t c a a a c a a a a c c a a c t a t t g c c g c c

8100 15 20 25 30 TGDVTVAPSNFANGVAEWIS MS2-N55K TGDVTVAPSNFANGVAEWIS g a c t g g c g a c g t t a c a g t a g c t c c a t c a a a t t t t g c g a a c g g a g t c g c t g a g t g g a t t a g c t g a c c g c t g c a a t g t c a t c g a g g t a g t t t a a a a c g c t t g c c t c a g c g a c t c a c c t a a t c

8160 35 40 45 50 SNSRSQAYKVTCSVRQSSAQ MS2-N55K SNSRSQAYKVTCSVRQSSAQ c t c a a a t t c a a g g t c c c a g g c c t a c a a g g t t a c c t g t t c t g t t a g g c a g a g t t c t g c g c a g a g t t t a a g t t c c a g g g t c c g g a t g t t c c a a t g g a c a a g a c a a t c c g t c t c a a g a c g c g t

8220 55 60 65 70 KRKYTIKVEVPKVATQTVGG MS2-N55K KRKYTIKVEVPKVATQTVGG a a a a a g a a a a t a t a c c a t c a a g g t t g a a g t c c c t a a a g t t g c a a c a c a a a c a g t c g g t g g t t t t t c t t t t a t a t g g t a g t t c c a a c t t c a g g g a t t t c a a c g t t g t g t t t g t c a g c c a c c

SspIEco53kISacI 8280 75 80 85 90 VELPVAAWRSYLNMELTIPI MS2-N55K VELPVAAWRSYLNMELTIPI t g t t g a g c t c c c t g t g g c a g c c t g g a g a t c t t a c t t a a a c a t g g a g c t a a c a a t t c c a a t a c a a c t c g a g g g a c a c c g t c g g a c c t c t a g a a t g a a t t t g t a c c t c g a t t g t t a a g g t t a

8340 95 100 105 110 FATNSDCELIVKAMQGLLKD MS2-N55K FATNSDCELIVKAMQGLLKD a t t c g c t a c a a a c t c t g a t t g t g a a c t g a t t g t t a a g g c g a t g c a a g g t c t c t t g a a a g a t a a g c g a t g t t t g a g a c t a a c a c t t g a c t a a c a a t t c c g c t a c g t t c c a g a g a a c t t t c t

8400 115 120 125 GNPIPSAIAANSGIY GSGSG MS2-N55K GNPIPSAIAANSGIYGSGSG t g g a a a c c c t a t a c c g t c c g c t a t c g c a g c t a a c a g c g g t a t c t a t g g c t c t g g a t c g g g a c c t t t g g g a t a t g g c a g g c g a t a g c g t c g a t t g t c g c c a t a g a t a c c g a g a c c t a g c c c

SpeI 8460 (in frame with MS2-N55K) SGSGSTSSGGS 5 10 TNLSDIIEK UGI SGSGSTSSGGSTNLSDIIEK g t c g g g t t c t g g c t c a a c t a g t t c a g g c g g g t c a a c a a a c c t c a g t g a c a t c a t a g a g a a c a g c c c a a g a c c g a g t t g a t c a a g t c c g c c c a g t t g t t t g g a g t c a c t g t a g t a t c t c t t

8520 15 20 25 30 ETGKQLVIQESILMLPEEVE UGI ETGKQLVIQESILMLPEEVE g g a a a c t g g t a a g c a a c t g g t t a t t c a a g a g a g c a t t c t g a t g c t c c c t g a g g a a g t c g a c c t t t g a c c a t t c g t t g a c c a a t a a g t t c t c t c g t a a g a c t a c g a g g g a c t c c t t c a g c t

8580 35 40 45 50 EVIGNKPESDILVHTAYDES UGI EVIGNKPESDILVHTAYDES g g a a g t t a t a g g a a a c a a g c c t g a g a g c g a t a t a c t c g t g c a t a c g g c g t a t g a c g a g a g c c t t c a a t a t c c t t t g t t c g g a c t c t c g c t a t a t g a g c a c g t a t g c c g c a t a c t g c t c t c

8640 55 60 65 70 TDENVMLLTSDAPEYKPWAL UGI TDENVMLLTSDAPEYKPWAL c a c g g a t g a a a a t g t g a t g c t c t t g a c c a g c g a t g c g c c a g a a t a t a a a c c c t g g g c a c t g t g c c t a c t t t t a c a c t a c g a g a a c t g g t c g c t a c g c g g t c t t a t a t t t g g g a c c c g t g a

8700 75 80 VIQDSNGENKIKML (in frame with UGI) SGGS PK UGI SV40 NLS VIQDSNGENKIKMLSGGSPK g g t t a t t c a g g a c t c t a a c g g g g a a a a t a a a a t c a a g a t g c t c t c a g g t g g c t c c c c a a a c c a a t a a g t c c t g a g a t t g c c c c t t t t a t t t t a g t t c t a c g a g a g t c c a c c g a g g g g t t t

Acc65IKpnI 8760 130 KKRKV GT 1 5 10 KRPAATKKAGQAK SV40 NLS nucleoplasmin NLS KKRKVGTKRPAATKKAGQAK g a a g a a a c g c a a g g t t g g t a c c a a g c g t c c t g c t g c c a c c a a a a a g g c c g g a c a g g c t a a c t t c t t t g c g t t c c a a c c a t g g t t c g c a g g a c g a c g g t g g t t t t t c c g g c c t g t c c g a t t

EagINotI 8820 15 KKK * nucleoplasmin NLS NOS terminator KKK* g a a a a a g a a g t a g t g g c g g c c g c c g a c g t c c g a t c g t t c a a a c a t t t g g c a a t a a a g t t t c t t t t t c t t c a t c a c c g c c g g c g g c t g c a g g c t a g c a a g t t t g t a a a c c g t t a t t t c a a a

BsaBI 8880 NOS terminator c t t a a g a t t g a a t c c t g t t g c c g g t c t t g c g a t g a t t a t c a t a t a a t t t c t g t t g a a t t a g a a t t c t a a c t t a g g a c a a c g g c c a g a a c g c t a c t a a t a g t a t a t t a a a g a c a a c t t a a t

NsiI 8940 NOS terminator c g t t a a g c a t g t a a t a a t t a a c a t g t a a t g c a t g a c g t t a t t t a t g a g a t g g g t t t t t a t g c a a t t c g t a c a t t a t t a a t t g t a c a t t a c g t a c t g c a a t a a a t a c t c t a c c c a a a a a t a

9000 NOS terminator g a t t a g a g t c c c g c a a t t a t a c a t t t a a t a c g c g a t a g a a a a c a a a a t a t a g c g c g c a a a c t a a t c t c a g g g c g t t a a t a t g t a a a t t a t g c g c t a t c t t t t g t t t t a t a t c g c g c g t t t

MauBI 9060 NOS terminator c t a g g a t a a a t t a t c g c g c g c g g t g t c a t c t a t g t t a c t a g a t c g g g a a t t g a t c c c c c c g a t c c t a t t t a a t a g c g c g c g c c a c a g t a g a t a c a a t g a t c t a g c c c t t a a c t a g g g g g g

BstZ17I 9120 attR5 t c g a c a g c t t c c g g a a a g g g c g a a t t c g c a a c t t t g t a t a c a a a a g t t g a a c g a g a a a c g a g c t g t c g a a g g c c t t t c c c g c t t a a g c g t t g a a a c a t a t g t t t t c a a c t t g c t c t t t g c

9180 attR5 t a a a a t g a t a t a a a t a t c a a t a t a t t a a a t t a g a t t t t g c a t a a a a a a c a g a c t a c a t a a a t t t t a c t a t a t t t a t a g t t a t a t a a t t t a a t c t a a a a c g t a t t t t t t g t c t g a t g t a t t

9240 attR5 T7 promoter G G G A T A T C A C T C T7 t a c t g t a a a a c a c a a c a t a t c c a g t c a c t a t g c c a t c c a g c t g a t a t c c c c t a t a g t g a g a t g a c a t t t t g t g t t g t a t a g g t c a g t g a t a c g g t a g g t c g a c t a t a g g g g a t a t c a c t c

9300 T7 promoter M13 rev A G C A T A A T T7 C A G T A T C G A C A A A G G A C M13 Reverse t c g t a t t a c a t g g t c a t a g c t g t t t c c t g g c a g c t c t g g c c c g t g t c t c a a a a t c t c t g a a g c a t a a t g t a c c a g t a t c g a c a a a g g a c c g t c g a g a c c g g g c a c a g a g t t t t a g a g a c t

3ʹ 5ʹ 9342 t g t t a c a t t g c a c a a g a t a a a a a t a t a t c a t c a t g c c t c c t c a c a a t g t a a c g t g t t c t a t t t t t a t a t a g t a g t a c g g a g g a g

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.