LOCUS Cre-IRES-PuroR. 9229 bp DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS Cre-IRES-PuroR. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9229) AUTHORS Somers A, Jean JC, Sommer CA, Omari A, Ford CC, Mills JA, Ying L, Sommer AG, Jean JM, Smith BW, Lafyatis R, Demierre MF, Weiss DJ, French DL, Gadue P, Murphy GJ, Mostoslavsky G, Kotton DN TITLE Generation of transgene-free lung disease-specific human induced pluripotent stem cells using a single excisable lentiviral stem cell cassette. JOURNAL Stem Cells. 2010 Oct . 28(10):1728-40. PUBMED 20715179 REFERENCE 2 (bases 1 to 9229) TITLE Direct Submission REFERENCE 3 (bases 1 to 9229) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells. 2010 Oct . 28(10):1728-40." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..9229 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 55..74 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 266..899 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 946..1071 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1568..1801 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1986..2030 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" CDS 2179..2220 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 2323..2440 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2488..3666 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" regulatory 3671..3680 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 3707..4735 /label=Cre /note="site-specific recombinase" misc_feature 4741..5319 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 5323..5919 /label=PuroR /note="puromycin N-acetyltransferase" misc_feature 5936..6524 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6672..6852 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(6899..6917) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6985..7089 /label=AmpR promoter CDS 7090..7947 /label=AmpR /note="beta-lactamase" rep_origin 8121..8709 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8863..8880 /label=L4440 /note="L4440 vector, forward primer" primer_bind 8972..8991 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 9120..9141 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 9156..9186 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9194..9210 /label=lac operator /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)." ORIGIN 1 atgacatgat tacgaatttc acaaataaag catttttttc actgcattct agttgtggtt 61 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc 121 aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc 181 atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt 241 aaatatgtac tacaaactta gtagttggaa gggctaattc actcccaaag aagacaagat 301 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 361 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 421 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 481 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 541 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 601 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 661 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 721 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 781 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 841 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 901 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg 961 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca 1021 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag 1081 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa 1141 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct 1201 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt 1261 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg 1321 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa 1381 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag 1441 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac 1501 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag 1561 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag 1621 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga 1681 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca 1741 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc 1801 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta 1861 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca 1921 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc 1981 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt 2041 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg 2101 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt 2161 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa 2221 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatctc 2281 gacggtatcg ccgaattcac aaatggcagt attcatccac aattttaaaa gaaaaggggg 2341 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac 2401 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag 2461 cagagatcca gtttggacta gtcgtgaggc tccggtgccc gtcagtgggc agagcgcaca 2521 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga 2581 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 2641 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 2701 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg 2761 ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat 2821 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct 2881 tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg 2941 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga 3001 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg 3061 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc 3121 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct 3181 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg 3241 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc 3301 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca 3361 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag 3421 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta 3481 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa 3541 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga 3601 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg 3661 tcgtgaagcg gccgccatgg gcacacgact ccccaagaag aggaaggtgt ccaatttact 3721 gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa 3781 gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct 3841 tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc 3901 cgcagaacct gaagatgttc gagattatct tctatatctt caggcgcgcg gtctggcagt 3961 aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc 4021 acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt 4081 tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt 4141 tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg 4201 gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat 4261 ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag 4321 caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat 4381 ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa 4441 tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt 4501 tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc 4561 ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc 4621 aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca tgaactatat 4681 ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattagag 4741 atccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt 4801 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac 4861 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc 4921 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa 4981 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg 5041 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg 5101 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc 5161 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat 5221 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg 5281 tggttttcct ttgaaaaaca cgatgataat atggccacac atatgaccga gtacaagccc 5341 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5401 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5461 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5521 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5581 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5641 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5701 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5761 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5821 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5881 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aaatcgatag atcctaatca 5941 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 6001 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 6061 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 6121 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 6181 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 6241 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 6301 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg 6361 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 6421 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 6481 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac 6541 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg 6601 aagggctaat tcactcccaa cgaagacaag atcacctgca ggacaggcgc gccctgcttt 6661 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac 6721 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg 6781 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga 6841 aaatctctag cacccgggcg attaaggaaa gggctagatc attcttgaag acgaaagggc 6901 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca 6961 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 7021 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 7081 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt 7141 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag 7201 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt 7261 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg 7321 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag 7381 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta 7441 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg 7501 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta 7561 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac 7621 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt 7681 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca 7741 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag 7801 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta 7861 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag 7921 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt 7981 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 8041 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 8101 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 8161 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 8221 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag 8281 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 8341 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 8401 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 8461 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 8521 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 8581 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 8641 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 8701 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 8761 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 8821 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 8881 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 8941 tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 9001 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 9061 agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt 9121 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt 9181 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagct //