LOCUS Exported 8579 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral EOS reporter with Sox2 enhancer (x4), expresses EGFP and Puro resistance. ACCESSION . VERSION . KEYWORDS PL-SIN-EOS-S(4+)-EiP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8579) 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 8579) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8579 /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" misc_feature 2201..2434 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." primer_bind 2763..2779 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 3756..3765 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3762..4481 /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 3762..4478 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3807..3828) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4068..4087) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4415..4436 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4528..5114 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4695..4712) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4922..4941 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5115..5714 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(5115..5134) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5611..5631 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 5856..6089 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6123..6139) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6123..6139) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6136..6158) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6147..6163 /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(6171..6201) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6216..6237 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6354..6371) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6525..7113) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6605..6624) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7284..8144) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7907..7926 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8145..8249) /gene="bla" /label=AmpR promoter rep_origin 8300..8435 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(8354..8373) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 8552..8570 /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 agaggccaat gaaggagaga 601 acaacagctt 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 cccgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 2101 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 2161 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 2221 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 2281 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 2341 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 2401 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 2461 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 2521 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttgg 2581 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt 2641 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa 2701 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcct 2761 ccctttatcc agccctcact ccttctctag gcgccggaat tcccagtcca agctaggcag 2821 gttcccctct aattaatgca gagactctaa aagaatttcc cgggctcggg cagccattgt 2881 gatgcatata ggattattca cgtggtaatg agcacagtcg acagttcttg ctctcgagta 2941 gaatcgaatt cccagtccaa gctaggcagg ttcccctcta attaatgcag agactctaaa 3001 agaatttccc gggctcgggc agccattgtg atgcatatag gattattcac gtggtaatga 3061 gcacagtcga cagttcttgc tctcgagtag aatcgaattc ccagtccaag ctaggcaggt 3121 tcccctctaa ttaatgcaga gactctaaaa gaatttcccg ggctcgggca gccattgtga 3181 tgcatatagg attattcacg tggtaatgag cacagtcgac agttcttgct ctcgagtaga 3241 atcgaattcc cagtccaagc taggcaggtt cccctctaat taatgcagag actctaaaag 3301 aatttcccgg gctcgggcag ccattgtgat gcatatagga ttattcacgt ggtaatgagc 3361 acagtcgaca gttcttgctc tcgagtagaa tcgaattctc ccctccccca gcctgaaacc 3421 tgcttgctca ggggtggagc ttcctgctca ttcgttctgc cacgcccact gctggaacct 3481 gcggagccac acccgtgcac ctttctactg gaccagagat tattcggcgg gaatcgggtc 3541 ccctccccct tccttcataa ctagtgtcgc aactataaaa tttgagcctt gatcagagta 3601 actgtcttgg ctacattctt tcttccgccc cgtctagatt cctctcttac agctcgagcg 3661 gccttctcag tcgaaccgtt cacgttgcga gctgctggcg gccgcaacaa gatctgcgat 3721 ctaagtaagc ttggcattcc ggtactgttg gtaaagccac catggtgagc aagggcgagg 3781 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 3841 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 3901 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct 3961 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 4021 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 4081 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 4141 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca 4201 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca 4261 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 4321 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg 4381 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 4441 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccaa ctcgacgtta 4501 ttcccttcga tcgaattccg cccccccccc ctctccctcc ccccccccta acgttactgg 4561 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 4621 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 4681 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 4741 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 4801 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 4861 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 4921 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 4981 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 5041 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata 5101 atatggccac aaccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg 5161 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca 5221 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc 5281 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct 5341 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg 5401 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc 5461 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg 5521 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg 5581 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct 5641 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca 5701 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac 5761 cccatgcatc gatttcgaac ccggggtacc tttaagacca atgacttaca aggcagctgt 5821 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaaag 5881 aagacaagat ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg 5941 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 6001 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 6061 cttttagtca gtgtggaaaa tctctagcag aattctagcg attctagctt ggcgtaatca 6121 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 6181 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 6241 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 6301 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 6361 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 6421 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 6481 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 6541 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6601 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6661 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 6721 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6781 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6841 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6901 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6961 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 7021 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 7081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 7141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 7201 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 7261 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 7321 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 7381 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 7441 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 7501 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 7561 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 7621 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7681 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7741 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7801 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7861 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7921 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7981 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 8041 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 8101 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 8161 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 8221 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggact 8281 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca 8341 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga 8401 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg 8461 gttgctgact aattgagatg agcttgcatg ccgacattga ttattgacta gtccctaaga 8521 aaccattctt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc //