LOCUS       V014827                 9745 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014827
VERSION     V014827
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9745)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1721..1765
                     /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             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2027..2144
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        complement(2528..2895)
                     /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"
     promoter        2912..3422
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3441..4184
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     misc_feature    4200..4786
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4806..5402
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    5430..5654
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    5668..6256
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6463..7096
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7225..7241)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7249..7265)
                     /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)."
     promoter        complement(7273..7303)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7318..7339)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7627..8212)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8386..9243)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9244..9348)
                     /label="AmpR promoter"
     polyA_signal    9396..9530
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgacttaact tgtttattgc agcttataat ggttacaaat
     2221 aaggcaatag catcacaaat ttcacaaata aggcattttt ttcactgcat tctagttttg
     2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctcaaatccc tcggaagctg
     2341 cgcctgtctt aggttggagt gatacatttt tatcactttt acccgtcttt ggattaggca
     2401 gtagctctga cggccctcct gtcttaggtt agtgaaaaat gtcactctct tacccgtcat
     2461 tggctgtcca gcttagctcg caggggaggt ggtctggatc cgccggcacc ggtgtatacg
     2521 ggaattcttt acgagggtag gaagtggtac ggaaagttgg tataagacaa aagtgttgtg
     2581 gaattgaagt ttactcaaaa aatcagcact cttttatagg cgccctggtt tacataagca
     2641 aagcttatac gttctctatc actgataggg agtaaactgg atatacgttc tctatcactg
     2701 atagggagta aactgtagat acgttctcta tcactgatag ggagtaaact ggtcatacgt
     2761 tctctatcac tgatagggag taaactcctt atacgttctc tatcactgat agggagtaaa
     2821 gtctgcatac gttctctatc actgataggg agtaaactct tcatacgttc tctatcactg
     2881 atagggagta aactcgaggt gataattcca cggggttggg gttgcgcctt ttccaaggca
     2941 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
     3001 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
     3061 ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct
     3121 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
     3181 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
     3241 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
     3301 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
     3361 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
     3421 agggggatca tcgaattacc atgtctagac tggacaagag caaagtcata aactctgctc
     3481 tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
     3541 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
     3601 ccctgccaat cgagatgctg gacaggcatc atacccactc ctgccccctg gaaggcgagt
     3661 catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc ctctcacatc
     3721 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
     3781 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
     3841 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
     3901 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg aaacaagcaa
     3961 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
     4021 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg
     4081 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
     4141 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaacg cgcgaatcgc
     4201 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     4261 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     4321 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     4381 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     4441 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     4501 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     4561 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     4621 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     4681 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     4741 gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccaca aggagacgac
     4801 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     4861 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     4921 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4981 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5041 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     5101 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     5161 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5221 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5281 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5341 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5401 cctagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     5461 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5521 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5581 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5641 ggtgggctct atggacgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
     5701 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5761 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5821 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5881 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5941 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6001 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6061 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6121 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6181 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6241 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa
     6301 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     6361 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     6421 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
     6481 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     6541 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     6601 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     6661 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     6721 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     6781 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     6841 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     6901 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6961 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7021 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7081 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     7141 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac cgtcgacctc
     7201 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     7261 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7321 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7381 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7441 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7501 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7561 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7981 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8041 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     8101 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8161 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     8221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     8281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     8341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     8401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     9001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     9061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     9121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     9181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     9241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     9301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     9361 aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg
     9421 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     9481 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa
     9541 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt
     9601 acctagtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
     9661 aattgtattt gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt
     9721 aaatatgtac tacaaactta gtagt
//
