LOCUS       V015004                10471 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015004
VERSION     V015004
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10471)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(216..350)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        398..502
                     /label="AmpR promoter"
     CDS             503..1360
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1534..2119
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2407..2428
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2443..2473
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     LTR             complement(2650..3283)
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     misc_feature    complement(3490..4078)
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(4092..4316)
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4344..4940)
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    complement(4960..5546)
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             complement(5562..6305)
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        complement(6324..6834)
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     promoter        6851..7218
                     /label="TRE3GS promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription
                     factors"
     CDS             7257..7973
                     /label="EGFP"
                     /note="enhanced GFP"
     polyA_signal    complement(8151..8285)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(8328..8445)
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(8517..8558)
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     CDS             complement(8707..8751)
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     misc_feature    complement(8936..9169)
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     misc_feature    complement(9666..9791)
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     LTR             complement(9838..10471)
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaaact actaagtttg
       61 tagtacatat ttaacaaata caatttcttt aaaatgaaaa taattcagag gaatcacagg
      121 tttagagtaa atgaaaccac taggtaattg gcagtggtaa tagggtatgg ggtgggaagt
      181 ttgggatgat tttggttagc ttgagttatc cagttgatcc agacatgata agatacattg
      241 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      301 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt gatctcccga
      361 tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1561 ctgctgcttg caaacaaaaa aaccgctacc agcggtggtt tgtttgccgg atcaagagct
     1621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2281 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2341 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2401 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2461 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2521 ccatgattac gccaagctct agctagaggt cgacggtaaa acgctagcaa tgtcaaggcc
     2581 tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata agatgacatg
     2641 aactactact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     2701 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     2761 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     2821 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     2881 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag
     2941 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga
     3001 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca
     3061 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta
     3121 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct
     3181 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga
     3241 tcaaggatat cttgtcttcg ttgggagtga attagccctt ccagtcccct cttttctttt
     3301 aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg tacctgaggt
     3361 gtgactggaa aacccacctc ctcctcctct tgtgcttcta gccaggcaca atcagcattg
     3421 gtagctgctg tattgctact tgtgattgct ccatgttttt ctaggtctcg actgcagaat
     3481 taattccagg cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa
     3541 gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga
     3601 gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga
     3661 gcagccatgg aaaggacgtc agcttccccg acaacaccac ggaattgtca gtgcccaaca
     3721 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag
     3781 ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag
     3841 tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca
     3901 actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac
     3961 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     4021 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt
     4081 tccagacgcg tccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
     4141 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     4201 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     4261 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     4321 ggctgatcag cgggtttaaa ctaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
     4381 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     4441 ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     4501 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg
     4561 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     4621 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     4681 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     4741 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     4801 gtgacccgct cgatgtggcg gtccgggtcg acggtgtggc gcgtggcggg gtagtcggcg
     4861 aacgcggcgg cgagggtgcg tacggcccgg gggacgtcgt cgcgggtggc gaggcgcacc
     4921 gtgggcttgt actcggtcat ggaaggtcgt ctccttgtgg gttgtggcaa gcttatcatc
     4981 gtgtttttca aaggaaaacc acgtccccgt ggttcggggg gcctagacgt ttttttaacc
     5041 tcgactaaac acatgtaaag catgtgcacc gaggccccag atcagatccc atacaatggg
     5101 gtaccttctg ggcatccttc agccccttgt tgaatacgct tgaggagagc catttgactc
     5161 tttccacaac tatccaactc acaacgtggc actggggttg tgccgccttt gcaggtgtat
     5221 cttatacacg tggcttttgg ccgcagaggc acctgtcgcc aggtgggggg ttccgctgcc
     5281 tgcaaagggt cgctacagac gttgtttgtc ttcaagaagc ttccagagga actgcttcct
     5341 tcacgacatt caacagacct tgcattcctt tggcgagagg ggaaagaccc ctaggaatgc
     5401 tcgtcaagaa gacagggcca ggtttccggg ccctcacatt gccaaaagac ggcaatatgg
     5461 tggaaaataa catatagaca aacgcacacc ggccttattc caagcggctt cggccagtaa
     5521 cgttaggggg gggggaggga gaggggcgat tcgcgcgttt acccggggag catgtcaagg
     5581 tcaaaatcgt caagagcgtc agcaggcagc atatcaaggt caaagtcgtc aagggcatcg
     5641 gctgggagca tgtctaagtc aaaatcgtca agggcgtcgg tcggcccgcc gctttcgcac
     5701 tttagctgtt tctccaggcc acatatgatt agttccaggc cgaaaaggaa ggcaggttcg
     5761 gctccctgcc ggtcgaacag ctcaattgct tgtttcagaa gtgggggcat agaatcggtg
     5821 gtaggtgtct ctctttcctc ttttgctact tgatgctcct gttcctccaa tacgcagccc
     5881 agtgtaaagt ggcccacggc ggacagagcg tacagtgcgt tctccaggga gaagccttgc
     5941 tgacacagga acgcgagctg attttccagg gtttcgtact gtttctctgt tgggcgggtg
     6001 ccgagatgca ctttagcccc gtcgcgatgt gagaggagag cacagcggta tgacttggcg
     6061 ttgttccgca gaaagtcttg ccatgactcg ccttccaggg ggcaggagtg ggtatgatgc
     6121 ctgtccagca tctcgattgg cagggcatcg agcagggccc gcttgttctt cacgtgccag
     6181 tacagggtag gctgctcaac tcccagcttt tgagcgagtt tccttgtcgt caggccttcg
     6241 ataccgactc cattgagtaa ttccagagca gagtttatga ctttgctctt gtccagtcta
     6301 gacatggtaa ttcgatgatc cccctgggga gagaggtcgg tgattcggtc aacgagggag
     6361 ccgactgccg acgtgcgctc cggaggcttg cagaatgcgg aacaccgcgc gggcaggaac
     6421 agggcccaca ctaccgcccc acaccccgcc tcccgcaccg ccccttcccg gccgctgctc
     6481 tcggcgcgcc ctgctgagca gccgctattg gccacagccc atcgcggtcg gcgcgctgcc
     6541 attgctccct ggcgctgtcc gtctgcgagg gtactagtga gacgtgcggc ttccgtttgt
     6601 cacgtccggc acgccgcgaa ccgcaaggaa ccttcccgac ttaggggcgg agcaggaagc
     6661 gtcgccgggg ggcccacaag ggtagcggcg aagatccggg tgacgctgcg aacggacgtg
     6721 aagaatgtgc gagacccagg gtcggcgccg ctgcgtttcc cggaaccacg cccagagcag
     6781 ccgcgtccct gcgcaaaccc agggctgcct tggaaaaggc gcaaccccaa ccccgtggaa
     6841 ttatcacctc gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta
     6901 tcagtgatag agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga
     6961 gtttactccc tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag
     7021 aacgtatcta cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta
     7081 tcagtgatag agaacgtata agctttgctt atgtaaacca gggcgcctat aaaagagtgc
     7141 tgattttttg agtaaacttc aattccacaa cacttttgtc ttataccaac tttccgtacc
     7201 acttcctacc ctcgtaaaga attcccgtat acaccggtgc cggcggatcc gccaccatgg
     7261 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     7321 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     7381 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     7441 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     7501 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     7561 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     7621 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     7681 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     7741 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     7801 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     7861 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     7921 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtagagac
     7981 cacctcccct gcgagctaag ctggacagcc aatgacgggt aagagagtga catttttcac
     8041 taacctaaga caggagggcc gtcagagcta ctgcctaatc caaagacggg taaaagtgat
     8101 aaaaatgtat cactccaacc taagacaggc gcagcttccg agggatttga gatccagaca
     8161 tgataagata cattgatgag tttggacaaa ccaaaactag aatgcagtga aaaaaatgcc
     8221 ttatttgtga aatttgtgat gctattgcct tatttgtaac cattataagc tgcaataaac
     8281 aagttaagtc gataaactgg atctctgctg tccctgtaat aaacccgaaa attttgaatt
     8341 tttgtaattt gtttttgtag ttctttagtt tgtatgtctg ttgctattat gtctactatt
     8401 ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaggcgat accgtcgaga
     8461 tccgttcact aatcgaatgg atctgtctct gtctctctct ccaccttctt cttctattcc
     8521 ttcgggcctg tcgggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
     8581 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
     8641 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
     8701 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
     8761 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
     8821 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
     8881 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
     8941 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
     9001 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
     9061 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
     9121 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
     9181 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
     9241 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
     9301 tccaggtctg aagatcagcg gccggccgct tgctgtgcgg tggtcttact tttgttttgc
     9361 tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca
     9421 caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg
     9481 gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagga
     9541 ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt
     9601 ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta
     9661 atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc
     9721 gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg
     9781 cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc
     9841 tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca
     9901 acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg
     9961 ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag
    10021 tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc cccagtcccg
    10081 cccaggccac gcctccctgg aaagtcccca gcggaaagtc ccttgtagca agctcgatat
    10141 cagcagttct tgaagtactc cggatgcagc tctcgggcca cgtgatgaaa tgctaggcgg
    10201 ctgtcaaacc tccactctaa cacttctctc tccgggtcat ccatcccatg caggctcaca
    10261 gggtgtaaca agctggtgtt ctctccttta ttggcctctt ctaccttatc tggctcaact
    10321 ggtactagct tgtagcacca tccaaaggtc agtggatatc tgacccctgg ccctggtgtg
    10381 tagttctgct aatcagggaa gtagccttgt gtgtggtaga tccacagatc aaggatatct
    10441 tgtcttcttt gggagtgaat tagcccttcc a
//
