LOCUS       V001194                 9229 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001194
VERSION     V001194
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."
            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"
                     /label="Protein Tat"
     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
//
