LOCUS Exported 7813 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector, expresses EGFP under Nanog promoter. ACCESSION . VERSION . KEYWORDS PL-SIN-Nanog-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7813) AUTHORS Hotta A, Cheung AY, Farra N, Vijayaragavan K, Seguin CA, Draper JS, Pasceri P, Maksakova IA, Mager DL, Rossant J, Bhatia M, Ellis J TITLE Isolation of human iPS cells using EOS lentiviral vectors to select for pluripotency. JOURNAL Nat Methods. 2009 May . 6(5):370-6. PUBMED 19404254 REFERENCE 2 (bases 1 to 7813) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7813 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" LTR 785..1042 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1693..1810 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" CDS 4254..4973 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4254..4970 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4299..4320) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4560..4579) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4907..4928 /label=EGFP-C /note="EGFP, forward primer" LTR 5100..5333 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(5370..5392) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5381..5397 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5405..5435) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5450..5471 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5588..5605) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5759..6347) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5839..5858) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7140..7159 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7378..7482) /gene="bla" /label=AmpR promoter promoter complement(7519..7715) /label=SV40 promoter /note="SV40 early promoter" rep_origin 7533..7668 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(7587..7606) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 7786..7804 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gcggccgctg gaagggctaa 421 tttggtccca aaaaagacaa gagatccttg atctgtggat ctaccacaca caaggctact 481 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat 541 ggtgcttcaa gttagtacca gttgaaccag agcaagtaga agaggccaaa taaggagaga 601 agaacagctt gttacaccct atgagccagc atgggatgga ggacccggag ggagaagtat 661 tagtgtggaa gtttgacagc ctcctagcat ttcgtcacat ggcccgagag ctgcatccgg 721 agtactacaa agactgctga catcgagctt tctacaaggg actttccgct ggggactttc 781 cagggaggtg tggcctgggc gggactgggg agtggcgagc cctcagatgc tacatataag 841 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc 901 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgctcaaa 961 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag 1021 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagtaaagc 1081 cagaggagat ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg 1141 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg 1201 ggtgcgagag cgtcggtatt aagcggggga gaattagata aatgggaaaa aattcggtta 1261 aggccagggg gaaagaaaca atataaacta aaacatatag tatgggcaag cagggagcta 1321 gaacgattcg cagttaatcc tggcctttta gagacatcag aaggctgtag acaaatactg 1381 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca 1441 atagcagtcc tctattgtgt gcatcaaagg atagatgtaa aagacaccaa ggaagcctta 1501 gataagatag aggaagagca aaacaaaagt aagaaaaagg cacagcaagc agcagctgac 1561 acaggaaaca acagccaggt cagccaaaat taccctatag tgcagaacct ccaggggcaa 1621 atggtacatc aggccatatc acctagaact ttaaattaag acagcagtac aaatggcagt 1681 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat 1741 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat 1801 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggaaag gaccagcaaa 1861 gctcctctgg aaaggtgaag gggcagtagt aatacaagat aatagtgaca taaaagtagt 1921 gccaagaaga aaagcaaaga tcatcaggga ttatggaaaa cagatggcag gtgatgattg 1981 tgtggcaagt agacaggatg aggattaaca catggaaaag attagtaaaa caccatagct 2041 ctagagcgat ccgatcttca gacctggagg aggcgatatg agggacaatt ggagaagtga 2101 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa 2161 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt 2221 cttgggagca gcaggaagca ctatgggctg cacgtcaatg acgctgacgg tacaggccag 2281 acaattattg tctgatatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca 2341 acagcatctg ttgcaactca cagtctgggg catcaaacag ctccaggcaa gaatcctggc 2401 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact 2461 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat 2521 ttggaataac atgacctgga tggagtggga cagagaaatt aacaattaca caagcttggt 2581 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc 2641 accattatcg tttcagaccc acctcccaat cccgagggga cccgacaggc ccgaaggaat 2701 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatccca 2761 aaagtcagct tgtgtgggag caaagccagc tgctctgggt gcagacccag gagcagagtg 2821 cagaggagaa tgagtcaaag agttttgtct tcaaaaatta cataatcggg atttgctaag 2881 agtttacttt tcggtatgga agactggaaa agagaaagaa atcttaggtt tcttgaatgt 2941 tgggtttggg aataggaagg aaaatcgaaa actgtagact ttgtccataa atgttagtgc 3001 tggaacccca ctctaaaaac tttgttcctt tggaaaacac ctcccttccc ccagaaacac 3061 acacacccac acgagatggg cacggagtag tcttgaaaga catgacaaat caccagacct 3121 gggaagaagc taaagagcca gagggaaaaa gccagaagtc gactacctgg gaggagggat 3181 agacaagaaa ccaaactaaa ggaaactaag gtaggtgctg aaaacaagta ccattttcaa 3241 cattaactga tgccttggct tcatgctata atgccatgtt gtgtttcact ataacctcag 3301 agtgaatgaa agaggaaaat ggagctagtt gaaatttctg cctaaactag ccagattttg 3361 agacactaag ttatctcaaa tcaagaaatc accctaatga gaatttcaat aacctcagga 3421 atttaaggtg catgcatccc ccaccccccc cttttttttt tgagacgtag tcccgctctg 3481 ttgcccaggc tggagtacag tggcgcgata tcggctcacc acaacctctg cctcccaggt 3541 tcaagggatt ctcccgcctc agcttccaga gtagctggga ctacagacac ccaccaccat 3601 gcgtggctaa tttttgtatt tttagtagag agggggtttc gccatgttgg ccaggctggt 3661 ttcaaactcc tgacttcagg tgatccgcct gccacggcct cccaatttac tgggattaca 3721 ggggtgggcc accgcgcccg gcctttttct taatttttaa aaatattaaa gttttatccc 3781 attcctgttg aaccatattc ctgatttaaa agttggaaac gtggtgaacc tagaagtatt 3841 tgttgctggg tttgtcttca ggttctgttg ctcggttttc tagttcccca cctagtctgg 3901 gttactctgc agctactttt gcattacaat ggccttggtg agactggtag acgggattaa 3961 ctgagaattc acaagggtgg gtcagtaggg ggtgtgcccg ccaggagggg tgggtctaag 4021 gtgatagagc cttcattata aatctagaga ctccaggatt ttaacgttct gctggactga 4081 gctggttgcc tcatgttatt atgcaggcaa ctcactttat cccaatttct tgatactttt 4141 ccttctggag gtcctatttc tctaacatct tccagaaaag tcttaaagct gccttaacct 4201 tttttccagt ccacctctta aattttttcc tcctcttcct ctatactaac accatggtga 4261 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg 4321 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc 4381 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga 4441 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg 4501 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg 4561 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc 4621 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg 4681 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca 4741 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact 4801 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga 4861 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg 4921 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc 4981 aacctcgacg ttattccctt cgcgaaggcc tatcgatttc gaacccgggg tacctttaag 5041 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 5101 ggaagggcta attcactccc aaagaagaca agatctgctt tttgcctgta ctgggtctct 5161 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 5221 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 5281 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagaattct 5341 agcttggcgt aatcatggtc atagctgttc ctgtgtgaaa ttgttatccg ctcacaattc 5401 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 5461 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 5521 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 5581 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 5641 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 5701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 5761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 5821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 5881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 5941 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 6001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 6061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 6121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 6181 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccacttacct 6241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 6301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 6361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 6421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 6481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 6541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 6601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 6661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 6721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 6781 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgtgccattg ctacaggcat 6841 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6901 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6961 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 7021 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 7081 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 7141 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7201 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7261 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7321 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 7381 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7441 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7501 gccacctggg actagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt 7561 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca 7621 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag 7681 gggcgggact atggttgctg actaattgag atgtagcttg catgccgaca tggattattg 7741 actagtccct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg 7801 aggccctttc gtc //