LOCUS       V013999                 9243 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013999
VERSION     V013999
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9243)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9243
                     /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(2544..2911)
                     /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        2928..3438
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3457..4200
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        4214..4543
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             4552..5148
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5165..5753
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5961..6594
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6723..6739)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6747..6763)
                     /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(6771..6801)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6816..6837)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7125..7710)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7884..8741)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8742..8846)
                     /label="AmpR promoter"
     polyA_signal    8894..9028
                     /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 cgctcgaatc gaagtccagc
     2521 aaccggtgta tacggaagaa ttctttacga gggtaggaag tggtacggaa agttggtata
     2581 agacaaaagt gttgtggaat tgaagtttac tcaaaaaatc agcactcttt tataggcgcc
     2641 ctggtttaca taagcaaagc ttatacgttc tctatcactg atagggagta aactggatat
     2701 acgttctcta tcactgatag ggagtaaact gtagatacgt tctctatcac tgatagggag
     2761 taaactggtc atacgttctc tatcactgat agggagtaaa ctccttatac gttctctatc
     2821 actgataggg agtaaagtct gcatacgttc tctatcactg atagggagta aactcttcat
     2881 acgttctcta tcactgatag ggagtaaact cgaggtgata attccacggg gttggggttg
     2941 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     3001 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     3061 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     3121 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     3181 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     3241 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
     3301 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     3361 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     3421 accgacctct ctccccaggg ggatcatcga attaccatgt ctagactgga caagagcaaa
     3481 gtcataaact ctgctctgga attactcaat ggagtcggta tcgaaggcct gacgacaagg
     3541 aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag
     3601 cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac ccactcctgc
     3661 cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc ataccgctgt
     3721 gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa
     3781 cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag
     3841 aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa
     3901 caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca
     3961 cttctgaaac aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc
     4021 ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg
     4081 accgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt
     4141 gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg
     4201 taaacgcgcg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4321 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4381 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4441 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4501 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaacgcgac catgaccgag
     4561 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     4621 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     4681 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4741 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4801 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4861 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4921 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4981 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5041 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5101 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aacgcgtctg
     5161 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5221 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5281 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5341 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5401 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5461 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5521 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     5581 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5641 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5701 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     5761 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     5821 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg tttttccagt
     5881 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     5941 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct
     6001 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     6061 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     6121 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     6181 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     6241 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     6301 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     6361 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     6421 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6481 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6541 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6601 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6661 agaggccttg acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca
     6721 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6781 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6841 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6901 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6961 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     7681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7861 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7921 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7981 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8041 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8101 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8161 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8221 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8281 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8341 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8401 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8461 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8521 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8581 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8641 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8701 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8761 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8821 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8881 ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     8941 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     9001 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
     9061 caaacttccc accccatacc ctattaccac tgccaattac ctagtggttt catttactct
     9121 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
     9181 ctacaaactt agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     9241 agt
//
