Addgene pLenti-HF1RA-P2A-GFP-PGK-Puro - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pLenti-HF1RA-P2A-GFP-PGK-Puro


Map View

M13 Reverse (12,370 .. 12,386) lac operator M13/pUC Reverse (12,351 .. 12,373) lac promoter CAP binding site L4440 (12,138 .. 12,155) AflIII - PciI (12,040) pBR322ori-F (11,885 .. 11,904) ori Amp-R (10,583 .. 10,602) AmpR promoter F1ori-F (10,076 .. 10,097) f1 ori F1ori-R (9866 .. 9885) M13/pUC Forward (9630 .. 9652) M13 Forward (9621 .. 9638) M13 fwd T7 (9592 .. 9611) T7 promoter AvrII (9571) SfiI (9524) SV40 ori EBV-rev (9367 .. 9386) SV40 poly(A) signal SV40pA-R (9313 .. 9332) SnaBI (9202) 3' LTR (ΔU3) WPRE WPRE-R (8358 .. 8378) XbaI (8291) CsiI - SexAI * (8257) Puro-F (8186 .. 8206) BstEII (7824) RsrII (7806) BsiWI (7746) PflFI - Tth111I (7732) Puro-R (7690 .. 7709) mPGK-F (7602 .. 7621) PGK promoter SpeI (7415) MSCV-rev (7205 .. 7228) BmtI (7163) NheI (7159) BsrGI (7148) EGFP-C (7092 .. 7113) M13 rev T3 (12,405 .. 12,425) T3 promoter RSV promoter 5' LTR (truncated) HIV-1 Ψ NruI * (611) NotI (922) BbvCI (1198) gp41 peptide cPPT/CTS PaeR7I - PspXI - XhoI (1883) EF-1α core promoter BamHI (2134) SV40 NLS PspOMI (3576) ApaI (3580) EcoRV (4074) TspMI - XmaI (4991) SmaI (4993) PmlI (5010) BsaBI * (5719) PshAI (6326) nucleoplasmin NLS P2A EXFP-R (6745 .. 6764) pLenti-HF1RA-P2A-GFP-PGK-Puro 12,448 bp

Sequence View

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

120 RSV promoter c a t g c c t t a c a a g g a g a g a a a a a g c a c c g t g c a t g c c g a t t g g t g g a a g t a a g g t g g t a c g t a c g g a a t g t t c c t c t c t t t t t c g t g g c a c g t a c g g c t a a c c a c c t t c a t t c c a c c a t g

180 RSV promoter g a t c g t g c c t t a t t a g g a a g g c a a c a g a c g g g t c t g a c a t g g a t t g g a c g a a c c a c t g a a c t a g c a c g g a a t a a t c c t t c c g t t g t c t g c c c a g a c t g t a c c t a a c c t g c t t g g t g a c t t

240 RSV promoter 5' LTR (truncated) t t g c c g c a t t g c a g a g a t a t t g t a t t t a a g t g c c t a g c t c g a t a c a t a a a c g g g t c t c t c a a c g g c g t a a c g t c t c t a t a a c a t a a a t t c a c g g a t c g a g c t a t g t a t t t g c c c a g a g a g

300 5' LTR (truncated) t g g t t a g a c c a g a t c t g a g c c t g g g a g c t c t c t g g c t a a c t a g g g a a c c c a c t g c t t a a g a c c a a t c t g g t c t a g a c t c g g a c c c t c g a g a g a c c g a t t g a t c c c t t g g g t g a c g a a t t c

360 5' LTR (truncated) c c t c a a t a a a g c t t g c c t t g a g t g c t t c a a g t a g t g t g t g c c c g t c t g t t g t g t g a c t c t g g a g t t a t t t c g a a c g g a a c t c a c g a a g t t c a t c a c a c a c g g g c a g a c a a c a c a c t g a g a

420 5' LTR (truncated) g g t a a c t a g a g a t c c c t c a g a c c c t t t t a g t c a g t g t g g a a a a t c t c t a g c a g t g g c g c c c c a t t g a t c t c t a g g g a g t c t g g g a a a a t c a g t c a c a c c t t t t a g a g a t c g t c a c c g c g g

480 HIV-1 Ψ c g a a c a g g g a c t t g a a a g c g a a a g g g a a a c c a g a g g a g c t c t c t c g a c g c a g g a c t c g g c g c t t g t c c c t g a a c t t t c g c t t t c c c t t t g g t c t c c t c g a g a g a g c t g c g t c c t g a g c c g

540 HIV-1 Ψ t t g c t g a a g c g c g c a c g g c a a g a g g c g a g g g g c g g c g a c t g g t g a g t a c g c c a a a a a t t t a a c g a c t t c g c g c g t g c c g t t c t c c g c t c c c c g c c g c t g a c c a c t c a t g c g g t t t t t a a a

600 HIV-1 Ψ t g a c t a g c g g a g g c t a g a a g g a g a g a g a t g g g t g c g a g a g c g t c a g t a t t a a g c g g g g g a a c t g a t c g c c t c c g a t c t t c c t c t c t c t a c c c a c g c t c t c g c a g t c a t a a t t c g c c c c c t

NruI* 660 g a a t t a g a t c g c g a t g g g a a a a a a t t c g g t t a a g g c c a g g g g g a a a g a a a a a a t a t a a a t c t t a a t c t a g c g c t a c c c t t t t t t a a g c c a a t t c c g g t c c c c c t t t c t t t t t t a t a t t t a

720 t a a a a c a t a t a g t a t g g g c a a g c a g g g a g c t a g a a c g a t t c g c a g t t a a t c c t g g c c t g t a t t t t g t a t a t c a t a c c c g t t c g t c c c t c g a t c t t g c t a a g c g t c a a t t a g g a c c g g a c a

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

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

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

NotI 960 MR g t a a g a c c a c c g c a c a g c a a g c g g c c g c t g a t c t t c a g a c c t g g a g g a g g a g a t a t g a g g c a t t c t g g t g g c g t g t c g t t c g c c g g c g a c t a g a a g t c t g g a c c t c c t c c t c t a t a c t c c

1020 DNWRSELYKYKVVKIEPLGV g a c a a t t g g a g a a g t g a a t t a t a t a a a t a t a a a g t a g t a a a a a t t g a a c c a t t a g g a g t a c t g t t a a c c t c t t c a c t t a a t a t a t t t a t a t t t c a t c a t t t t t a a c t t g g t a a t c c t c a t

1080 RRE APTKAKRRVVQREKRAVGIG g c a c c c a c c a a g g c a a a g a g a a g a g t g g t g c a g a g a g a a a a a a g a g c a g t g g g a a t a g g a c g t g g g t g g t t c c g t t t c t c t t c t c a c c a c g t c t c t c t t t t t t c t c g t c a c c c t t a t c c t

1140 RRE ALFLGFLGAAGSTMGAASMT g c t t t g t t c c t t g g g t t c t t g g g a g c a g c a g g a a g c a c t a t g g g c g c a g c g t c a a t g a c g c g a a a c a a g g a a c c c a a g a a c c c t c g t c g t c c t t c g t g a t a c c c g c g t c g c a g t t a c t g c

BbvCI 1200 RRE LTVQARQLLSGIVQQQNNLL c t g a c g g t a c a g g c c a g a c a a t t a t t g t c t g g t a t a g t g c a g c a g c a g a a c a a t t t g c t g g a c t g c c a t g t c c g g t c t g t t a a t a a c a g a c c a t a t c a c g t c g t c g t c t t g t t a a a c g a c

1260 RRE RAIEAQQHLLQLTVWGIKQL a g g g c t a t t g a g g c g c a a c a g c a t c t g t t g c a a c t c a c a g t c t g g g g c a t c a a g c a g c t c t c c c g a t a a c t c c g c g t t g t c g t a g a c a a c g t t g a g t g t c a g a c c c c g t a g t t c g t c g a g

1320 RRE QARILAVERYLKDQQLLGIW c a g g c a a g a a t c c t g g c t g t g g a a a g a t a c c t a a a g g a t c a a c a g c t c c t g g g g a t t t g g g t c c g t t c t t a g g a c c g a c a c c t t t c t a t g g a t t t c c t a g t t g t c g a g g a c c c c t a a a c c

1380 GCSGKLICTTAVPWNASWSN g g t t g c t c t g g a a a a c t c a t t t g c a c c a c t g c t g t g c c t t g g a a t g c t a g t t g g a g t a a t c c a a c g a g a c c t t t t g a g t a a a c g t g g t g a c g a c a c g g a a c c t t a c g a t c a a c c t c a t t a

1440 KSLEQIWNHTTWMEWDREIN a a a t c t c t g g a a c a g a t t t g g a a t c a c a c g a c c t g g a t g g a g t g g g a c a g a g a a a t t a a c t t t a g a g a c c t t g t c t a a a c c t t a g t g t g c t g g a c c t a c c t c a c c c t g t c t c t t t a a t t g

1500 1 KN gp41 peptide NYTSLIHSLIEESQNQQEKN a a t t a c a c a a g c t t a a t a c a c t c c t t a a t t g a a g a a t c g c a a a a c c a g c a a g a a a a g a a t t t a a t g t g t t c g a a t t a t g t g a g g a a t t a a c t t c t t a g c g t t t t g g t c g t t c t t t t c t t a

1560 5 10 15 EQELLELDKWASL (in frame with gp41 peptide) WNWFNIT gp41 peptide EQELLELDKWASLWNWFNIT g a a c a a g a a t t a t t g g a a t t a g a t a a a t g g g c a a g t t t g t g g a a t t g g t t t a a c a t a a c a c t t g t t c t t a a t a a c c t t a a t c t a t t t a c c c g t t c a a a c a c c t t a a c c a a a t t g t a t t g t

1620 (in frame with gp41 peptide) NWLWYIKLFIMIVGGLVGLR NWLWYIKLFIMIVGGLVGLR a a t t g g c t g t g g t a t a t a a a a t t a t t c a t a a t g a t a g t a g g a g g c t t g g t a g g t t t a a g a t t a a c c g a c a c c a t a t a t t t t a a t a a g t a t t a c t a t c a t c c t c c g a a c c a t c c a a a t t c t

1680 (in frame with gp41 peptide) IVFAVLSIVNRVRQGYSPLS IVFAVLSIVNRVRQGYSPLS a t a g t t t t t g c t g t a c t t t c t a t a g t g a a t a g a g t t a g g c a g g g a t a t t c a c c a t t a t c g t a t c a a a a a c g a c a t g a a a g a t a t c a c t t a t c t c a a t c c g t c c c t a t a a g t g g t a a t a g c

1740 (in frame with gp41 peptide) FQTHLPTPRGPRPQF* cPPT/CTS FQTHLPTPRGPRPQF* t t t c a g a c c c a c c t c c c a a c c c c g a g g g g a c c c a g a c c a c a a t t t t a a a a g a a a a g g g g g a a a g t c t g g g t g g a g g g t t g g g g c t c c c c t g g g t c t g g t g t t a a a a t t t t c t t t t c c c c c

1800 cPPT/CTS g a t t g g g g g g t a c a g t g c a g g g g a a a g a a t a g t a g a c a t a a t a g c a a c a g a c a t a c a a a c c t a a c c c c c c a t g t c a c g t c c c c t t t c t t a t c a t c t g t a t t a t c g t t g t c t g t a t g t t t g

1860 cPPT/CTS *LSL t a a a g a a t t a c a a a a a c a a a t t a c a a a a a t t c a a a a t t t t c g g g t t t a t t a c a g g g a c a g a t t t c t t a a t g t t t t t g t t t a a t g t t t t t a a g t t t t a a a a g c c c a a a t a a t g t c c c t g t c

PaeR7IPspXIXhoI 1920 EF-1 α core promoter LSGSQRRSSHSRHGDTPLAC c a g a g a t c c a c t t t g g c g c c g g c t c g a g t g g c t c c g g t g c c c g t c a g t g g g c a g a g c g c a g t c t c t a g g t g a a a c c g c g g c c g a g c t c a c c g a g g c c a c g g g c a g t c a c c c g t c t c g c g t

1980 EF-1 α core promoter MAWLGRSTPPLPRCNFRHRS c a t c g c c c a c a g t c c c c g a g a a g t t g g g g g g a g g g g t c g g c a a t t g a a c c g g t g c c t a g a g t a g c g g g t g t c a g g g g c t c t t c a a c c c c c c t c c c c a g c c g t t a a c t t g g c c a c g g a t c t

2040 EF-1 α core promoter FTARPLSPFHHRTSAGGKER g a a g g t g g c g c g g g g t a a a c t g g g a a a g t g a t g t c g t g t a c t g g c t c c g c c t t t t t c c c g c t t c c a c c g c g c c c c a t t t g a c c c t t t c a c t a c a g c a c a t g a c c g a g g c g g a a a a a g g g c

2100 EF-1 α core promoter PHPLVTYLHLLRRSREKECR a g g g t g g g g g a g a a c c g t a t a t a a g t g c a g t a g t c g c c g t g a a c g t t c t t t t t c g c a a c g t c c c a c c c c c t c t t g g c a t a t a t t c a c g t c a t c a g c g g c a c t t g c a a g a a a a a g c g t t g c

BamHI 2160 1 DYKD EF-1 α core promoter FLAG Kozak sequence MDYKD TQRWFVPTTVRSGGGHIVFV g g t t t g c c g c c a g a a c a c a g g t g t c g t g a c g c g g g a t c c g c c a c c a t g g a t t a c a a a g a c c c a a a c g g c g g t c t t g t g t c c a c a g c a c t g c g c c c t a g g c g g t g g t a c c t a a t g t t t c t g

2220 5 DDDK MA 130 PKKKRKV (in frame with SV40 NLS) GIHGVPA FLAG SV40 NLS DDDKMAPKKKRKVGIHGVPA IVILHGWLLLPLDTDVSDWC 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 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 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 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

2280 A 5 10 15 20 DKKYSIGLDIGTNSVGWAV Cas9 ADKKYSIGLDIGTNSVGWAV GVLLVADAQVDAGVRHAPGH 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 a c c a a c t c t g t g g g c t g g g c c g t g 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 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

2340 25 30 35 40 ITDEYKVPSKKFKVLGNTDR Cas9 ITDEYKVPSKKFKVLGNTDR DGVLVLHGALFELHQAVGVP 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 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 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 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

2400 45 50 55 60 HSIKKNLIGALLFDSGETAE Cas9 HSIKKNLIGALLFDSGETAE VADLLVQDSGQQEVAAFCGL 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 g a c a g c g g c g a a a c a g c c g a g 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 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

2460 65 70 75 80 ATRLKRTARRRYTRRKNRIC Cas9 ATRLKRTARRRYTRRKNRIC GGPQLSGGSSSVGSPLVPDA 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 c c a g a c g g a a g a a c c g g a t c t g 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 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

2520 85 90 95 100 YLQEIFSNEMAKVDDSFFHR Cas9 YLQEIFSNEMAKVDDSFFHR IQLLDEAVLHGLHVVAEEVS 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 g t g g a c g a c a g c t t c t t c c a c a g a 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 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

2580 105 110 115 120 LEESFLVEEDKKHERHPIFG Cas9 LEESFLVEEDKKHERHPIFG QFLGEQHFLILLVLPVGDEA 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 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 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 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

2640 125 130 135 140 NIVDEVAYHEKYPTIYHLRK Cas9 NIVDEVAYHEKYPTIYHLRK VDHVLHGVVLLVGGDVVQSL 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 c c c a c c a t c t a c c a c c t g a g a a a 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 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

2700 145 150 155 160 KLVDSTDKADLRLIYLALAH Cas9 KLVDSTDKADLRLIYLALAH FQHVAGVLGVQPQDIQGQGV 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 c t g a t c t a t c t g g c c c t g g c c c a 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 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

2760 165 170 175 180 MIKFRGHFLIEGDLNPDNSD Cas9 MIKFRGHFLIEGDLNPDNSD HDLEPAVEQDLAVQVGVVAV 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 c t g a a c c c c g a c a a c a g c g a c 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 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

2820 185 190 195 200 VDKLFIQLVQTYNQLFEENP Cas9 VDKLFIQLVQTYNQLFEENP HVLQEDLQHLGVVLQELFVG 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 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 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 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

2880 205 210 215 220 INASGVDAKAILSARLSKSR Cas9 INASGVDAKAILSARLSKSR DVGAAHVGLGDQRGSQALAS 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 c t g c c a g a c t g a g c a a g a g c a g a 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 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

2940 225 230 235 240 RLENLIAQLPGEKKNGLFGN Cas9 RLENLIAQLPGEKKNGLFGN PQFIQDGLQGALLLIAQESV 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 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 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 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

3000 245 250 255 260 LIALSLGLTPNFKSNFDLAE Cas9 LIALSLGLTPNFKSNFDLAE QNGQAQAQGGVELAVEVQGL 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 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 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 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

3060 265 270 275 280 DAKLQLSKDTYDDDLDNLLA Cas9 DAKLQLSKDTYDDDLDNLLA IGFQLQALVGVVVVQVVQQG 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 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 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 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

3120 285 290 295 300 QIGDQYADLFLAAKNLSDAI Cas9 QIGDQYADLFLAAKNLSDAI LDAVLVGVQKQGGLVQGVGD 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 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 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 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

3180 305 310 315 320 LLSDILRVNTEITKAPLSAS Cas9 LLSDILRVNTEITKAPLSAS QQAVDQSHVGLDGLGGQAGR 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 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 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 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

3240 325 330 335 340 MIKRYDEHHQDLTLLKALVR Cas9 MIKRYDEHHQDLTLLKALVR HDLSVVLVVLVQGQQFSEHP 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 a c c c t g c t g a a a g c t c t c g t g c g g 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 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

3300 345 350 355 360 QQLPEKYKEIFFDQSKNGYA Cas9 QQLPEKYKEIFFDQSKNGYA LLQRLLVFLNEEVLALVAVG 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 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 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 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

3360 365 370 375 380 GYIDGGASQEEFYKFIKPIL Cas9 GYIDGGASQEEFYKFIKPIL AVNVASGALFLEVLEDLGDQ 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 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 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 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

3420 385 390 395 400 EKMDGTEELLVKLNREDLLR Cas9 EKMDGTEELLVKLNREDLLR FLHVAGLFQEHLQVSLVQQP 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 c t g a a c a g a g a g g a c c t g c t g c g g 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 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

3480 405 410 415 420 KQRTFDNGSIPHQIHLGELH Cas9 KQRTFDNGSIPHQIHLGELH LLPGEVVAADGVLDVQSLQV 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 c a g a t c c a c c t g g g a g a g c t g c a 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 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

3540 425 430 435 440 AILRRQEDFYPFLKDNREKI Cas9 AILRRQEDFYPFLKDNREKI GNQPPLFIKVWEQLVVPFLD 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 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 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 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

PspOMIApaI 3600 445 450 455 460 EKILTFRIPYYVGPLARGNS Cas9 EKILTFRIPYYVGPLARGNS LLDQGEADGVVHARQGPSVA 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 g c c c t c t g g c c a g g g g a a a c a g c 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 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

3660 465 470 475 480 RFAWMTRKSEETITPWNFEE Cas9 RFAWMTRKSEETITPWNFEE SEGPHGSLALFGDGGPVELF 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 a t c a c c c c c t g g a a c t t c g a g g a a 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 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

3720 485 490 495 500 VVDKGASAQSFIERMTNFDK Cas9 VVDKGASAQSFIERMTNFDK HHVLASGGLAEDLPHGVEIL 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 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 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 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

3780 505 510 515 520 NLPNEKVLPKHSLLYEYFTV Cas9 NLPNEKVLPKHSLLYEYFTV VQGVLLHQGLVAQQVLVEGH 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 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 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 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

3840 525 530 535 540 YNELTKVKYVTEGMRKPAFL Cas9 YNELTKVKYVTEGMRKPAFL IVLQGFHFVHGLSHSLGGEQ 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 g a a t g a g a a a g c c c g c c t t c c t g 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 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

3900 545 550 555 560 SGEQKKAIVDLLFKTNRKVT Cas9 SGEQKKAIVDLLFKTNRKVT AALLFLGDHVQQELGVPFHG 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 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 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 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

3960 565 570 575 580 VKQLKEDYFKKIECFDSVEI Cas9 VKQLKEDYFKKIECFDSVEI HLLQFLVVELFDLAEVGHFD 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 g a g t g c t t c g a c t c c g t g g a a a t c 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 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

4020 585 590 595 600 SGVEDRFNASLGTYHDLLKI Cas9 SGVEDRFNASLGTYHDLLKI GAHFIPEVGGQACVVIQQFN 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 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 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 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

EcoRV 4080 605 610 615 620 IKDKDFLDNEENEDILEDIV Cas9 IKDKDFLDNEENEDILEDIV DLVLVEQVILFVLVNQFIDH 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 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 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 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

4140 625 630 635 640 LTLTLFEDREMIEERLKTYA Cas9 LTLTLFEDREMIEERLKTYA *A QGQCQKLVSLHDLFPQFGIG 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 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 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 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

4200 645 650 655 660 HLFDDKVMKQLKRRRYTGWG Cas9 HLFDDKVMKQLKRRRYTGWG WRNSSLTIFCSFRRLYVPQP VQEVVFHHLLQLPPSVGAPA 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 g g c g g a g a t a c a c c g g c t g g g g c 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 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

4260 665 670 675 680 ALSRKLINGIRDKQSGKTIL Cas9 ALSRKLINGIRDKQSGKTIL ARLRFSILPMRSLCDPLVIR GQAPLQDVADPVLLGALCDQ g c c 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 a a g c a g t c c g g c a a g a c a a t c c t g c g g 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 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

4320 685 690 695 700 DFLKSDGFANRNFMALIHDD Cas9 DFLKSDGFANRNFMALIHDD SKRFDSPKALLFKMARIWSS IEQLGVAEGVSVEHGQDVVV 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 c t t c a t g g c c c t g a t c c a c g a c 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 g a a g t a c c g g g a c t a g g t g c t g c t g

4380 705 710 715 720 SLTFKEDIQKAQVSGQGDSL Cas9 SLTFKEDIQKAQVSGQGDSL LRVKLSSMWFAWTDPWPSLR AQGKFLVDLFGLHGALAIAQ 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 g t g t c c g g c c a g g g c g a t a g c 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 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

4440 725 730 735 740 HEHIANLAGSPAIKKGILQT Cas9 HEHIANLAGSPAIKKGILQT CSCMALRAPLGAMLFPMRCV VLVNGIQGAAGGNLLADQLC 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 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 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 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

4500 745 750 755 760 VKVVDELVKVMGRHKPENIV Cas9 VKVVDELVKVMGRHKPENIV TFTTSSSTFTIPRCLGSFMT HLHHVLEHFHHAPVLGLVDH 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 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 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 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

4560 765 770 775 780 IEMARENQTTQKGQKNSRER Cas9 IEMARENQTTQKGQKNSRER ISIALSFWVVWFPCFFLRSL DFHGSLVLGGLLSLLVAALS 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 g a c a g a a g a a c a g c c g c g a g a g a 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 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

4620 785 790 795 800 MKRIEEGIKELGSQILKEHP Cas9 MKRIEEGIKELGSQILKEHP IFRISSPM HLPDFLADFLQAALDQFFVG 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 a g c c a g a t c c t g a a a g a a c a c c c c 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 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

4680 805 810 815 820 VENTQLQNEKLYLYYLQNGR Cas9 VENTQLQNEKLYLYYLQNGR HFVGLQLVLLQVQVVQLIPP 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 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 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 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

4740 825 830 835 840 DMYVDQELDINRLSDYDVDH Cas9 DMYVDQELDINRLSDYDVDH IHVHVLFQVDVPQGVVIHVM 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 c t g t c c g a c t a c g a t g t g g a c c a t 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 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

4800 845 850 855 860 IVPQSFLKDDSIDNKVLTRS Cas9 IVPQSFLKDDSIDNKVLTRS 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 a c a a g g t g c t g a c c a g a a g c 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 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

4860 865 870 875 880 DKNRGKSDNVPSEEVVKKMK Cas9 DKNRGKSDNVPSEEVVKKMK 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 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 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 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

4920 885 890 895 900 NYWRQLLNAKLITQRKFDNL Cas9 NYWRQLLNAKLITQRKFDNL 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 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 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 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

4980 905 910 915 920 TKAERGGLSELDKAGFIKRQ Cas9 TKAERGGLSELDKAGFIKRQ 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 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 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 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

PmlITspMIXmaISmaI 5040 925 930 935 940 LVETRAITKHVAQILDSRMN Cas9 LVETRAITKHVAQILDSRMN c t g g t g g a a a c c c g g g c c a t c a c a a a g c a c g t g g c 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 g a c c a c c t t t g g g c c c g g t a g t g t t t c g t g c a c c g 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

5100 945 950 955 960 TKYDENDKLIREVKVITLKS Cas9 TKYDENDKLIREVKVITLKS 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 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 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 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

5160 965 970 975 980 KLVSDFRKDFQFYKVREINN Cas9 KLVSDFRKDFQFYKVREINN 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 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 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 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

5220 985 990 995 1000 YHHAHDAYLNAVVGTALIKK Cas9 YHHAHDAYLNAVVGTALIKK 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 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 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 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

5280 1005 1010 1015 1020 YPKLESEFVYGDYKVYDVRK Cas9 YPKLESEFVYGDYKVYDVRK 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 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 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 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

5340 1025 1030 1035 1040 MIAKSEQEIGKATAKYFFYS Cas9 MIAKSEQEIGKATAKYFFYS 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 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 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 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

5400 1045 1050 1055 1060 NIMNFFKTEITLANGEIRKR Cas9 NIMNFFKTEITLANGEIRKR 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 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 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 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

5460 1065 1070 1075 1080 PLIETNGETGEIVWDKGRDF Cas9 PLIETNGETGEIVWDKGRDF 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 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 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 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

5520 1085 1090 1095 1100 ATVRKVLSMPQVNIVKKTEV Cas9 ATVRKVLSMPQVNIVKKTEV 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 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 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 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

5580 1105 1110 1115 1120 QTGGFSKESILPKRNSDKLI Cas9 QTGGFSKESILPKRNSDKLI 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 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 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 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

5640 1125 1130 1135 1140 ARKKDWDPKKYGGFDSPTVA Cas9 ARKKDWDPKKYGGFDSPTVA 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 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 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 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

5700 1145 1150 1155 1160 YSVLVVAKVEKGKSKKLKSV Cas9 YSVLVVAKVEKGKSKKLKSV 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 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 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 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

BsaBI* 5760 1165 1170 1175 1180 KELLGITIMERSSFEKNPID Cas9 KELLGITIMERSSFEKNPID 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 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 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 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

5820 1185 1190 1195 1200 FLEAKGYKEVKKDLIIKLPK Cas9 FLEAKGYKEVKKDLIIKLPK 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 g a c c t g a t c a t c a a g c t g c c t a a 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 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

5880 1205 1210 1215 1220 YSLFELENGRKRMLASAGEL Cas9 YSLFELENGRKRMLASAGEL 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 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 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 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

5940 1225 1230 1235 1240 QKGNELALPSKYVNFLYLAS Cas9 QKGNELALPSKYVNFLYLAS 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 g t g a a c t t c c t g t a c c t g g c c a g 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 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

6000 1245 1250 1255 1260 HYEKLKGSPEDNEQKQLFVE Cas9 HYEKLKGSPEDNEQKQLFVE 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 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 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 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

6060 1265 1270 1275 1280 QHKHYLDEIIEQISEFSKRV Cas9 QHKHYLDEIIEQISEFSKRV 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 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 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 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

6120 1285 1290 1295 1300 ILADANLDKVLSAYNKHRDK Cas9 ILADANLDKVLSAYNKHRDK 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 g c c t a c a a c a a g c a c c g g g a t a a g 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 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

6180 1305 1310 1315 1320 PIREQAENIIHLFTLTNLGA Cas9 PIREQAENIIHLFTLTNLGA 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 t t t a c c c t g a c c a a t c t g g g a g c 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 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

6240 1325 1330 1335 1340 PAAFKYFDTTIDRKRYTSTK Cas9 PAAFKYFDTTIDRKRYTSTK 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 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 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 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

6300 1345 1350 1355 1360 EVLDATLIHQSITGLYETRI Cas9 EVLDATLIHQSITGLYETRI 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 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 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 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

PshAI 6360 1365 DLSQLGGD 1 5 10 KRPAATKKAGQA Cas9 nucleoplasmin NLS DLSQLGGDKRPAATKKAGQA 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 g c g t c c t g c t g c t a c t a a g a a a g c t g g t c a a g c t 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 c g c a g g a c g a c g a t g a t t c t t t c g a c c a g t t c g a

6420 15 KKKK GSG 1 5 10 ATNFSLLKQAGDV nucleoplasmin NLS P2A KKKKGSGATNFSLLKQAGDV a a g a a a a a g a a a g g c a g c g g c g c c a c c a a c t t c a g c c t g c t g a a g c a g g c c g g c g a c g t g t t c t t t t t c t t t c c g t c g c c g c g g t g g t t g a a g t c g g a c g a c t t c g t c c g g c c g c t g c a c

6480 15 EENPGP 1 M 1a V 5 10 SKGEELFTGVVP P2A EGFP EENPGPMVSKGEELFTGVVP g a g g a g a a c c c c g g c c c c 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 c g g g g t g g t g c c c c t c c t c t t g g g g c c g g g g 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 g c c c c a c c a c g g g

6540 15 20 25 30 ILVELDGDVNGHKFSVSGEG EGFP ILVELDGDVNGHKFSVSGEG 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 t c t g g c g a g g g c 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 c a g a c c g c t c c c g

6600 35 40 45 50 EGDATYGKLTLKFICTTGKL EGFP EGDATYGKLTLKFICTTGKL 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 c c g g c a a g c t 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 g t g g c c g t t c g a c

6660 55 60 65 70 PVPWPTLVTTLTYGVQCFSR EGFP PVPWPTLVTTLTYGVQCFSR 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 c t t c a g c c g 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 c a c g a a g t c g g c g

6720 75 80 85 90 YPDHMKQHDFFKSAMPEGYV EGFP YPDHMKQHDFFKSAMPEGYV 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 g a a g g c t a c g t c 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 g c t t c c g a t g c a g

6780 95 100 105 110 QERTIFFKDDGNYKTRAEVK EGFP C T G C T G C C G T T G A T G T T C T G EXFP-R QERTIFFKDDGNYKTRAEVK 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 c g c c g a g g t g a 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 g c g g c t c c a c t t c

6840 115 120 125 130 FEGDTLVNRIELKGIDFKED EGFP FEGDTLVNRIELKGIDFKED 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 c t t c a a g g a g g a 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 g a a g t t c c t c c t g

6900 135 140 145 150 GNILGHKLEYNYNSHNVYIM EGFP GNILGHKLEYNYNSHNVYIM 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 c g t c t a t a t c a 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 g c a g a t a t a g t a c

6960 155 160 165 170 ADKQKNGIKANFKIRHNIED EGFP ADKQKNGIKANFKIRHNIED 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 c g a a c t t c a a g a t c c g c c a c a a c a t c g a g g 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 g c t t g a a g t t c t a g g c g g t g t t g t a g c t c c t g

7020 175 180 185 190 GSVQLADHYQQNTPIGDGPV EGFP GSVQLADHYQQNTPIGDGPV 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 c g a c g g c c c c g 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 g c t g c c g g g g c a c

7080 195 200 205 210 LLPDNHYLSTQSALSKDPNE EGFP LLPDNHYLSTQSALSKDPNE 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 a g a c c c c a a c g a g 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 t c t g g g g t t g c t c

7140 215 220 225 230 KRDHMVLLEFVTAAGITLGM 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 KRDHMVLLEFVTAAGITLGM 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 c a c t c t c g g c a t g 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 g t g a g a g c c g t a c

BmtINheIBsrGI 7200 235 DELYK* EGFP PGK promoter DELYK* g a c g a g c t g t a c a a g t a a g c t a g c c t a c c g g g t a g g g g a g g c g c t t t t c c c a a g g c a g t c c t g c t c g a c a t g t t c a t t c g a t c g g a t g g c c c a t c c c c t c c g c g a a a a g g g t t c c g t c a g

7260 PGK promoter C G T A C G C G A A A T C G T C G G G G C G A C MSCV-rev t g g a g c a t g c g c t t t a g c a g c c c c g c t g g g c a c t t g g c g c t a c a c a a g t g g c c t c t g g c c a c c t c g t a c g c g a a a t c g t c g g g g c g a c c c g t g a a c c g c g a t g t g t t c a c c g g a g a c c g g

7320 PGK promoter t c g c a c a c a t t c c a c a t c c a c c g g t a g g c g c c a a c c g g c t c c g t t c t t t g g t g g c c c c t t a g c g t g t g t a a g g t g t a g g t g g c c a t c c g c g g t t g g c c g a g g c a a g a a a c c a c c g g g g a a

7380 PGK promoter c g c g c c a c c t t c t a c t c c t c c c c t a g t c a g g a a g t t c c c c c c c g c c c c g c a g c t c g c g t c g c g c g g t g g a a g a t g a g g a g g g g a t c a g t c c t t c a a g g g g g g g c g g g g c g t c g a g c g c a g

SpeI 7440 PGK promoter g t g c a g g a c g t g a c a a a t g g a a g t a g c a c g t c t c a c t a g t c t c g t g c a g a t g g a c a g c a c c a c g t c c t g c a c t g t t t a c c t t c a t c g t g c a g a g t g a t c a g a g c a c g t c t a c c t g t c g t g

7500 PGK promoter MEAGRPLGQRPIAALLL c g c t g a g c a a t g g a a g c g g g t a g g c c t t t g g g g c a g c g g c c a a t a g c a g c t t t g c t c c t t g c g a c t c g t t a c c t t c g c c c a t c c g g a a a c c c c g t c g c c g g t t a t c g t c g a a a c g a g g a a

7560 PGK promoter RFLGSEAGKGWVRGRAQGRA c g c t t t c t g g g c t c a g a g g c t g g g a a g g g g t g g g t c c g g g g g c g g g c t c a g g g g c g g g c t g c g a a a g a c c c g a g t c t c c g a c c c t t c c c c a c c c a g g c c c c c g c c c g a g t c c c c g c c c g a

7620 PGK promoter C A T T C T G C A C G C T T C A A A A mPGK-F QGRGGRPKVLRRPGILHASK c a g g g g c g g g g c g g g c g c c c g a a g g t c c t c c g g a g g c c c g g c a t t c t g c a c g c t t c a a a a g t c c c c g c c c c g c c c g c g g g c t t c c a g g a g g c c t c c g g g c c g t a a g a c g t g c g a a g t t t t

7680 PGK promoter G mPGK-F AHVCRAVLLFLISGPFDLQP g c g c a c g t c t g c c g c g c t g t t c t c c t c t t c c t c a t c t c c g g g c c t t t c g a c c t g c a g c c c c g c g t g c a g a c g g c g c g a c a a g a g g a g a a g g a g t a g a g g c c c g g a a a g c t g g a c g t c g g g

Tth111IPflFI 7740 1 5 10 15 MTEYKPTVRLATRDDVP PuroR T A C T G G C T C A T G T T C G G G T G Puro-R KLTMTEYKPTVRLATRDDVP a a g c t t a c c a t g a c c g a g t a c a a g c c c a c g g t g c g c c t c g c c a c c c g c g a c g a c g t c c c c t t c g a a t g g t a c t g g c t c a t g t t c g g g t g c c a c g c g g a g c g g t g g g c g c t g c t g c a g g g g

BsiWI 7800 20 25 30 35 RAVRTLAAAFADYPATRHTV PuroR RAVRTLAAAFADYPATRHTV a g g g c c g t a c g c a c c c t c g c c g c c g c g t t c g c c g a c t a c c c c g c c a c g c g c c a c a c c g t c t c c c g g c a t g c g t g g g a g c g g c g g c g c a a g c g g c t g a t g g g g c g g t g c g c g g t g t g g c a g

BstEIIRsrII 7860 40 45 50 55 DPDRHIERVTELQELFLTRV PuroR DPDRHIERVTELQELFLTRV g a t c c g g a c c g c c a c a t c g a g c g g g t c a c c g a g c t g c a a g a a c t c t t c c t c a c g c g c g t c c t a g g c c t g g c g g t g t a g c t c g c c c a g t g g c t c g a c g t t c t t g a g a a g g a g t g c g c g c a g

7920 60 65 70 75 GLDIGKVWVADDGAAVAVWT PuroR GLDIGKVWVADDGAAVAVWT g g g c t c g a c a t c g g c a a g g t g t g g g t c g c g g a c g a c g g c g c c g c g g t g g c g g t c t g g a c c c c c g a g c t g t a g c c g t t c c a c a c c c a g c g c c t g c t g c c g c g g c g c c a c c g c c a g a c c t g g

7980 80 85 90 95 TPESVEAGAVFAEIGPRMAE PuroR TPESVEAGAVFAEIGPRMAE a c g c c g g a g a g c g t c g a a g c g g g g g c g g t g t t c g c c g a g a t c g g c c c g c g c a t g g c c g a g t g c g g c c t c t c g c a g c t t c g c c c c c g c c a c a a g c g g c t c t a g c c g g g c g c g t a c c g g c t c

8040 100 105 110 115 LSGSRLAAQQQMEGLLAPHR PuroR LSGSRLAAQQQMEGLLAPHR t t g a g c g g t t c c c g g c t g g c c g c g c a g c a a c a g a t g g a a g g c c t c c t g g c g c c g c a c c g g a a c t c g c c a a g g g c c g a c c g g c g c g t c g t t g t c t a c c t t c c g g a g g a c c g c g g c g t g g c c

8100 120 125 130 135 PKEPAWFLATVGVSPDHQGK PuroR PKEPAWFLATVGVSPDHQGK c c c a a g g a g c c c g c g t g g t t c c t g g c c a c c g t c g g c g t c t c g c c c g a c c a c c a g g g c a a g g g g t t c c t c g g g c g c a c c a a g g a c c g g t g g c a g c c g c a g a g c g g g c t g g t g g t c c c g t t c

8160 140 145 150 155 GLGSAVVLPGVEAAERAGVP PuroR GLGSAVVLPGVEAAERAGVP g g t c t g g g c a g c g c c g t c g t g c t c c c c g g a g t g g a g g c g g c c g a g c g c g c c g g g g t g c c c c c a g a c c c g t c g c g g c a g c a c g a g g g g c c t c a c c t c c g c c g g c t c g c g c g g c c c c a c g g g

8220 160 165 170 175 AFLETSAPRNLPFYERLGFT PuroR G C A A C C T C C C C T T C T A C G A G C Puro-F AFLETSAPRNLPFYERLGFT g c c t t c c t g g a g a c c t c c g c g c c c c g c a a c c t c c c c t t c t a c g a g c g g c t c g g c t t c a c c c g g a a g g a c c t c t g g a g g c g c g g g g c g t t g g a g g g g a a g a t g c t c g c c g a g c c g a a g t g g

SexAI*CsiI 8280 180 185 190 195 VTADVEVPEGPRTWCMTRKP PuroR VTADVEVPEGPRTWCMTRKP g t c a c c g c c g a c g t c g a g g t g c c c g a a g g a c c g c g c a c c t g g t g c a t g a c c c g c a a g c c c c a g t g g c g g c t g c a g c t c c a c g g g c t t c c t g g c g c g t g g a c c a c g t a c t g g g c g t t c g g g

XbaI 8340 200 GA* PuroR WPRE GA* g g t g c c t g a t t c t a g a a a g t c g a c a a t c a a c c t c t g g a t t a c a a a a t t t g t g a a a g a t t g c c a c g g a c t a a g a t c t t t c a g c t g t t a g t t g g a g a c c t a a t g t t t t a a a c a c t t t c t a a c

8400 WPRE A C A A C G A G G A A A A T G C G A T A C WPRE-R MP a c t g g t a t t c t t a a c t a t g t t g c t c c t t t t a c g c t a t g t g g a t a c g c t g c t t t a a t g c c t t g a c c a t a a g a a t t g a t a c a a c g a g g a a a a t g c g a t a c a c c t a t g c g a c g a a a t t a c g g a

8460 WPRE LYHAIASRMAFIFSSLYKSW MLLLPVWLSFSPPCINPG t t g t a t c a t g c t a t t g c t t c c c g t a t g g c t t t c a t t t t c t c c t c c t t g t a t a a a t c c t g g a a c a t a g t a c g a t a a c g a a g g g c a t a c c g a a a g t a a a a g a g g a g g a a c a t a t t t a g g a c c

8520 WPRE LLSLYEELWPVVRQRGVVCT CCLFMRSCGPLSGNVAWCAL t t g c t g t c t c t t t a t g a g g a g t t g t g g c c c g t t g t c a g g c a a c g t g g c g t g g t g t g c a c t a a c g a c a g a g a a a t a c t c c t c a a c a c c g g g c a a c a g t c c g t t g c a c c g c a c c a c a c g t g a

8580 WPRE VFADATPTGWGIATTCQLLS CLLTQPPLVGALPPPVSSFP g t g t t t g c t g a c g c a a c c c c c a c t g g t t g g g g c a t t g c c a c c a c c t g t c a g c t c c t t t c c c a c a a a c g a c t g c g t t g g g g g t g a c c a a c c c c g t a a c g g t g g t g g a c a g t c g a g g a a a g g

8640 WPRE GTFAFPLPIATAELIAACLA GLSLSPSLLPRRNSSPPALP g g g a c t t t c g c t t t c c c c c t c c c t a t t g c c a c g g c g g a a c t c a t c g c c g c c t g c c t t g c c c c c t g a a a g c g a a a g g g g g a g g g a t a a c g g t g c c g c c t t g a g t a g c g g c g g a c g g a a c g g

8700 WPRE RCWTGARLLGTDNSVVLSGK AAGQGLGCWALTIPWCCRGN c g c t g c t g g a c a g g g g c t c g g c t g t t g g g c a c t g a c a a t t c c g t g g t g t t g t c g g g g a a a g c g a c g a c c t g t c c c c g a g c c g a c a a c c c g t g a c t g t t a a g g c a c c a c a a c a g c c c c t t t

8760 WPRE (in frame with Factor Xa site) *RGKRPQEGTNGGPNQAPRG SSSFPWLLACVATWILRGTS HRPFLGCSPVLPPGFCAGRP t c a t c g t c c t t t c c t t g g c t g c t c g c c t g t g t t g c c a c c t g g a t t c t g c g c g g g a c g t c c a g t a g c a g g a a a g g a a c c g a c g a g c g g a c a c a a c g g t g g a c c t a a g a c g c g c c c t g c a g g

8820 WPRE EAVDR 1 RGEI Factor Xa site FCYVPSALNPADLPSRGLLP SATSLRPSIQRTFLPAACCR t t c t g c t a c g t c c c t t c g g c c c t c a a t c c a g c g g a c c t t c c t t c c c g c g g c c t g c t g c c g a a g a c g a t g c a g g g a a g c c g g g a g t t a g g t c g c c t g g a a g g a a g g g c g c c g g a c g a c g g c

8880 WPRE ALRPLPRLRLRPQTSRISLW LCGLFRVFAFALRRVGSPFG g c t c t g c g g c c t c t t c c g c g t c t t c g c c t t c g c c c t c a g a c g a g t c g g a t c t c c c t t t g g c g a g a c g c c g g a g a a g g c g c a g a a g c g g a a g c g g g a g t c t g c t c a g c c t a g a g g g a a a c c

8940 WPRE AASPRRL* PPPRVDFKTNDLQGSCRS* g c c g c c t c c c c g c g t c g a c t t t a a g a c c a a t g a c t t a c a a g g c a g c t g t a g a t c t t a g c c c g g c g g a g g g g c g c a g c t g a a a t t c t g g t t a c t g a a t g t t c c g t c g a c a t c t a g a a t c g g

9000 3' LTR ( Δ U3) a c t t t t t a a a a g a a a a g g g g g g a c t g g a a g g g c t a a t t c a c t c c c a a c g a a g a c a a g a t c t g a a a a a t t t t c t t t t c c c c c c t g a c c t t c c c g a t t a a g t g a g g g t t g c t t c t g t t c t a g

9060 3' LTR ( Δ U3) t g c t t t t t g c t t g t a c t g g g t c t c t c t g g t t a g a c c a g a t c t g a g c c t g g g a g c t c t c t g a c g a a a a a c g a a c a t g a c c c a g a g a g a c c a a t c t g g t c t a g a c t c g g a c c c t c g a g a g a c

9120 3' LTR ( Δ U3) g c t a a c t a g g g a a c c c a c t g c t t a a g c c t c a a t a a a g c t t g c c t t g a g t g c t t c a a g t a g c g a t t g a t c c c t t g g g t g a c g a a t t c g g a g t t a t t t c g a a c g g a a c t c a c g a a g t t c a t c

9180 3' LTR ( Δ U3) t g t g t g c c c g t c t g t t g t g t g a c t c t g g t a a c t a g a g a t c c c t c a g a c c c t t t t a g t c a g a c a c a c g g g c a g a c a a c a c a c t g a g a c c a t t g a t c t c t a g g g a g t c t g g g a a a a t c a g t c

SnaBI 9240 3' LTR ( Δ U3) t g t g g a a a a t c t c t a g c a g t a c g t a t a g t a g t t c a t g t c a t c t t a t t a t t c a g t a t t t a t a c a c c t t t t a g a g a t c g t c a t g c a t a t c a t c a a g t a c a g t a g a a t a a t a a g t c a t a a a t a

9300 SV40 poly(A) signal a a c t t g c a a a g a a a t g a a t a t c a g a g a g t g a g a g g 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 t t g a a c g t t t c t t t a c t t a t a g t c t c t c a c t c t c c 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

9360 SV40 poly(A) signal C G T T A T C G T A G T G T T T A A A G SV40pA-R g t t a c a a a t a a a g c a a 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 a a t g t t t a t t t c g t t 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

9420 SV40 poly(A) signal G T G G T T T G T C C A A A C T C A T C EBV-rev c t a g t t g t g g t t t g t c c a a a c t c a t c a a t g t a t c t t a t c a t g t c t g g c t c t a g c t a t c c c g a t c a a c a c c a a a c a g g t t t g a g t a g t t a c a t a g a a t a g t a c a g a c c g a g a t c g a t a g g g

9480 SV40 ori g c c c c t a a c t c c g c c c a t c c c g c c c c t a a c t c c g c c c a g t t c c g c c c a t t c t c c g c c c c a c g g g g a t t g a g g c g g g t a g g g c g g g g a t t g a g g c g g g t c a a g g c g g g t a a g a g g c g g g g t

SfiI 9540 SV40 ori t g g c t g a c t a a t t t t t t t t a t t t a t g c a g a g g c c g a g g c c g c c t c g g c c t c t g a g c t a t t a c c g a c t g a t t a a a a a a a a t a a a t a c g t c t c c g g c t c c g g c g g a g c c g g a g a c t c g a t a a

AvrII 9600 SV40 ori T7 promoter G G G A T A T C A T7 c c a g a a g t a g t g a g g a g g c t t t t t t g g a g g c c t a g g g a c g t a c c c a a t t c g c c c t a t a g t g g t c t t c a t c a c t c c t c c g a a a a a a c c t c c g g a t c c c t g c a t g g g t t a a g c g g g a t a t c a

9660 T7 promoter M13 fwd C T C A G C A T A A T T7 T G A C C G G C A G C A A A A T G T M13 Forward G C A A A A T G T T G C A G C A C T G A C C C M13/pUC Forward g a g t c g t a t t a c g c g c g c t c a c t g g c c g t c g t t t t a c a a c g t c g t g a c t g g g a a a a c c c t c t c a g c a t a a t g c g c g c g a g t g a c c g g c a g c a a a a t g t t g c a g c a c t g a c c c t t t t g g g a

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

9780 f1 ori g a a g a g g c c c g c a c c g a t c g c c c t t c c c a a c a g t t g c g c a g c c t g a a t g g c g a a t g g g a c c t t c t c c g g g c g t g g c t a g c g g g a a g g g t t g t c a a c g c g t c g g a c t t a c c g c t t a c c c t g

9840 f1 ori g c g c c c t g t a g c g g c g c a t t a a g c g c g g c g g g t g t g g t g g t t a c g c g c a g c g t g a c c g c t c g c g g g a c a t c g c c g c g t a a t t c g c g c c g c c c a c a c c a c c a a t g c g c g t c g c a c t g g c g a

9900 f1 ori G A G G A A A G C G A A A G A A G G G A F1ori-R a c a c t t g c c a g c g c c c t a g c g c c c g c t c c t t t c g c t t t c t t c c c t t c c t t t c t c g c c a c g t g t g a a c g g t c g c g g g a t c g c g g g c g a g g a a a g c g a a a g a a g g g a a g g a a a g a g c g g t g c

9960 f1 ori t t c g c c g g c t t t c c c c g t c a a g c t c t a a a t c g g g g g c t c c c t t t a g g g t t c c g a t t t a g t a a g c g g c c g a a a g g g g c a g t t c g a g a t t t a g c c c c c g a g g g a a a t c c c a a g g c t a a a t c a

10,020 f1 ori g c t t t a c g g c a c c t c g a c c c c a a a a a a c t t g a t t a g g g t g a t g g t t c a c g t a g t g g g c c a c g a a a t g c c g t g g a g c t g g g g t t t t t t g a a c t a a t c c c a c t a c c a a g t g c a t c a c c c g g t

10,080 f1 ori G T G G A F1ori-F t c g c c c t g a t a g a c g g t t t t t c g c c c t t t g a c g t t g g a g t c c a c g t t c t t t a a t a g t g g a a g c g g g a c t a t c t g c c a a a a a g c g g g a a a c t g c a a c c t c a g g t g c a a g a a a t t a t c a c c t

10,140 f1 ori C T C T T G T T C C A A A C T G G F1ori-F c t c t t g t t c c a a a c t g g a a c a a c a c t c a a c c c t a t c t c g g t c t a t t c t t t t g a t t t a t a a g a g a a c a a g g t t t g a c c t t g t t g t g a g t t g g g a t a g a g c c a g a t a a g a a a a c t a a a t a t t

10,200 f1 ori g g g a t t t t g c c g a t t t c g g c c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a a a a t t t a a c c c c t a a a a c g g c t a a a g c c g g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t t t t t a a a t t g

10,260 f1 ori AmpR promoter g c g a a t t t t a a c a a a a t a t t a a c g c t t a c a a t t t a g g t g g c a c t t t t c g g g g a a a t g t g c c g c t t a a a a t t g t t t t a t a a t t g c g a a t g t t a a a t c c a c c g t g a a a a g c c c c t t t a c a c g

10,320 AmpR promoter g c g g a a c c c c t a t t t g t t t a t t t t t c t a a a t a c a t t c a a a t a t g t a t c c g c t c a t g a g a c c g c c t t g g g g a t a a a c a a a t a a a a a g a t t t a t g t a a g t t t a t a c a t a g g c g a g t a c t c t g

10,380 1 5 signal sequence MSIQH AmpR promoter AmpR MSIQH a a t a a c c c t g a t a a a t g c t t c a a t a a t a t t g a a a a a g g a a g a g t a t g a g t a t t c a a c a t t t t a t t g g g a c t a t t t a c g a a g t t a t t a t a a c t t t t t c c t t c t c a t a c t c a t a a g t t g t a a

10,440 10 15 20 signal sequence FRVALIPFFAAFCLPVFA 25 HP AmpR FRVALIPFFAAFCLPVFAHP t c c g t g t c g c c c t t a t t c c c t t t t t t g c g g c a t t t t g c c t t c c t g t t t t t g c t c a c c c a g a g g c a c a g c g g g a a t a a g g g a a a a a a c g c c g t a a a a c g g a a g g a c a a a a a c g a g t g g g t c

10,500 30 35 40 45 ETLVKVKDAEDQLGARVGYI AmpR ETLVKVKDAEDQLGARVGYI a a a c g c t g g t g a a a g t a a a a g a t g c t g a a g a t c a g t t g g g t g c a c g a g t g g g t t a c a t c g t t t g c g a c c a c t t t c a t t t t c t a c g a c t t c t a g t c a a c c c a c g t g c t c a c c c a a t g t a g c

10,560 50 55 60 65 ELDLNSGKILESFRPEERFP AmpR ELDLNSGKILESFRPEERFP a a c t g g a t c t c a a c a g c g g t a a g a t c c t t g a g a g t t t t c g c c c c g a a g a a c g t t t t c c a a t t g a c c t a g a g t t g t c g c c a t t c t a g g a a c t c t c a a a a g c g g g g c t t c t t g c a a a a g g t t

10,620 70 75 80 85 MMSTFKVLLCGAVLSRIDAG AmpR C G A T A C A C C G C G C C A T A A T A Amp-R MMSTFKVLLCGAVLSRIDAG t g a t g a g c a c t t t t a a a g t t c t g c t a t g t g g c g c g g t a t t a t c c c g t a t t g a c g c c g g g c a c t a c t c g t g a a a a t t t c a a g a c g a t a c a c c g c g c c a t a a t a g g g c a t a a c t g c g g c c c g

10,680 90 95 100 105 QEQLGRRIHYSQNDLVEYSP AmpR QEQLGRRIHYSQNDLVEYSP a a g a g c a a c t c g g t c g c c g c a t a c a c t a t t c t c a g a a t g a c t t g g t t g a g t a c t c a c c a g t t c t c g t t g a g c c a g c g g c g t a t g t g a t a a g a g t c t t a c t g a a c c a a c t c a t g a g t g g t c

10,740 110 115 120 125 VTEKHLTDGMTVRELCSAAI AmpR VTEKHLTDGMTVRELCSAAI t c a c a g a a a a g c a t c t t a c g g a t g g c a t g a c a g t a a g a g a a t t a t g c a g t g c t g c c a t a a a g t g t c t t t t c g t a g a a t g c c t a c c g t a c t g t c a t t c t c t t a a t a c g t c a c g a c g g t a t t

10,800 130 135 140 145 TMSDNTAANLLLTTIGGPKE AmpR TMSDNTAANLLLTTIGGPKE c c a t g a g t g a t a a c a c t g c g g c c a a c t t a c t t c t g a c a a c g a t c g g a g g a c c g a a g g a g c g g t a c t c a c t a t t g t g a c g c c g g t t g a a t g a a g a c t g t t g c t a g c c t c c t g g c t t c c t c g

10,860 150 155 160 165 LTAFLHNMGDHVTRLDRWEP AmpR LTAFLHNMGDHVTRLDRWEP t a a c c g c t t t t t t g c a c a a c a t g g g g g a t c a t g t a a c t c g c c t t g a t c g t t g g g a a c c g g a t t g g c g a a a a a a c g t g t t g t a c c c c c t a g t a c a t t g a g c g g a a c t a g c a a c c c t t g g c c

10,920 170 175 180 185 ELNEAIPNDERDTTMPVAMA AmpR ELNEAIPNDERDTTMPVAMA a g c t g a a t g a a g c c a t a c c a a a c g a c g a g c g t g a c a c c a c g a t g c c t g t a g c a a t g g c a a t c g a c t t a c t t c g g t a t g g t t t g c t g c t c g c a c t g t g g t g c t a c g g a c a t c g t t a c c g t t

10,980 190 195 200 205 TTLRKLLTGELLTLASRQQL AmpR TTLRKLLTGELLTLASRQQL c a a c g t t g c g c a a a c t a t t a a c t g g c g a a c t a c t t a c t c t a g c t t c c c g g c a a c a a t t a a g t t g c a a c g c g t t t g a t a a t t g a c c g c t t g a t g a a t g a g a t c g a a g g g c c g t t g t t a a t t

11,040 210 215 220 225 IDWMEADKVAGPLLRSALPA AmpR IDWMEADKVAGPLLRSALPA t a g a c t g g a t g g a g g c g g a t a a a g t t g c a g g a c c a c t t c t g c g c t c g g c c c t t c c g g c t g a t c t g a c c t a c c t c c g c c t a t t t c a a c g t c c t g g t g a a g a c g c g a g c c g g g a a g g c c g a c

11,100 230 235 240 245 GWFIADKSGAGERGSRGIIA AmpR GWFIADKSGAGERGSRGIIA g c t g g t t t a t t g c t g a t a a a t c t g g a g c c g g t g a g c g t g g g t c t c g c g g t a t c a t t g c a g c g a c c a a a t a a c g a c t a t t t a g a c c t c g g c c a c t c g c a c c c a g a g c g c c a t a g t a a c g t c

11,160 250 255 260 265 ALGPDGKPSRIVVIYTTGSQ AmpR ALGPDGKPSRIVVIYTTGSQ c a c t g g g g c c a g a t g g t a a g c c c t c c c g t a t c g t a g t t a t c t a c a c g a c g g g g a g t c a g g g t g a c c c c g g t c t a c c a t t c g g g a g g g c a t a g c a t c a a t a g a t g t g c t g c c c c t c a g t c c

11,220 270 275 280 285 ATMDERNRQIAEIGASLIKH AmpR ATMDERNRQIAEIGASLIKH c a a c t a t g g a t g a a c g a a a t a g a c a g a t c g c t g a g a t a g g t g c c t c a c t g a t t a a g c a t t g t t g a t a c c t a c t t g c t t t a t c t g t c t a g c g a c t c t a t c c a c g g a g t g a c t a a t t c g t a a

11,280 W* AmpR W* g g t a a c t g t c a g a c c a a g t t t a c t c a t a t a t a c t t t a g a t t g a t t t a a a a c t t c a t t t t t c c a t t g a c a g t c t g g t t c a a a t g a g t a t a t a t g a a a t c t a a c t a a a t t t t g a a g t a a a a a

11,340 a a t t t a a a a g g a t c t a g g t g a a g a t c c t t t t t g a t a a t c t c a t g a c c a a a a t c c c t t a a c t t a a a t t t t c c t a g a t c c a c t t c t a g g a a a a a c t a t t a g a g t a c t g g t t t t a g g g a a t t g

11,400 ori g t g a g t t t t c g t t c 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 c a c t c a a a a g c a a 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

11,460 ori a t c c t t t t t t t c t g 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 a g g a a a a a a a g a c 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

11,520 ori t g g t t t g t t t g c c g 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 a c c a a a c a a a c g g c 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

11,580 ori g a g c g c a g a t a c c a 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 c t c g c g t c t a t g g t 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

11,640 ori a c t c t g t a g c a c c g 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 t g a g a c a t c g t g g c 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

11,700 ori g t g g c g a t a a g t c g 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 c a c c g c t a t t c a g c 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

11,760 ori a g c g g t c g g g c t g a 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 t c g c c a g c c c g a c t 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

11,820 ori c c g a a c t g a g a t a c 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 g g c t t g a c t c t a t g 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

11,880 ori a g g c g g a c a g g t a t 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 t c c g c c t g t c c a t a 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

11,940 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F c a g g g g g a a a c g c c 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 c c c c t t t g c g g 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

12,000 ori g t c g a t t t t t g t g a 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 a g c t a a a a a c a c t 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

AflIIIPciI 12,060 c c t t t t t a c g g t t c 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 g g a a a a a t g c c a a 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

12,120 c c c c t g a t t c t g t g 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 g g g a c t a a g a c a c 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

12,180 A G C G A G T C A G T G A G C G A G L4440 g c c g a a c g a c c g a g 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 c g g c t t g c t g g c t 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

12,240 a a c c g c c t c t c c c c 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 t t g g c g g a g a g g g g 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

12,300 CAP binding site a c t g g a a a g c g g g c a g t g a g c g c a a c g c a a t t a a t g t g a g t t a g c t c a c t c a t t a g g c a c t g a c c t t t c g c c c g t c a c t c g c g t t g c g t t a a t t a c a c t c a a t c g a g t g a g t a a t c c g t g

12,360 -35 -10 lac promoter lac operator A G C G G A T A A C M13/pUC Reverse c c c a g g c t t t a c a c t t t a t g c t t c c g g c t c g t a t g t t g t g t g g a a t t g t g a g c g g a t a a c g g g t c c g a a a t g t g a a a t a c g a a g g c c g a g c a t a c a a c a c a c c t t a a c a c t c g c c t a t t g

12,420 lac operator M13 rev T3 promoter A A T T T C A C A C A G G M13/pUC Reverse C A G G A A A C A G C T A T G A C M13 Reverse G C A A T T A A C C C T C A C T T3 a a t t t c a c a c a g g a a a c a g c t a t g a c c a t g a t t a c g c c a a g c g c g c a a t t a a c c c t c a c t t t a a a g t g t g t c c t t t g t c g a t a c t g g t a c t a a t g c g g t t c g c g c g t t a a t t g g g a g t g a

3ʹ 5ʹ 12,448 T3 promoter A A A G G T3 a a a g g g a a c a a a a g c t g g a g c t g c a a g c t t t c c c t t g t t t t c g a c c t c g a c g t t c 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.