LOCUS       V013512                 9806 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013512
VERSION     V013512
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9806)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9806
                     /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"
     enhancer        2201..2504
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    2802..2867
                     /label="MCS"
                     /note="multiple cloning site"
     CDS             2871..2927
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             2928..4574
                     /label="luciferase"
                     /note="firefly luciferase"
     promoter        4594..5093
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             5114..5710
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5727..6315
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6522..7155
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7284..7300)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7308..7324)
                     /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(7332..7362)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7377..7398)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7686..8274)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8448..9305)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9306..9410)
                     /label="AmpR promoter"
     polyA_signal    9458..9592
                     /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 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccatg gccaccaact
     2881 tcagcctgct gaagcaggcc ggcgacgtgg aggagaaccc cggccccgaa gatgccaaaa
     2941 acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc
     3001 tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac
     3061 atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag
     3121 ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct
     3181 tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta
     3241 acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg
     3301 tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca
     3361 tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca
     3421 ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct
     3481 tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca
     3541 agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca
     3601 tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg
     3661 gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt
     3721 accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc
     3781 tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc
     3841 taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg
     3901 ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa
     3961 ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg
     4021 tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc
     4081 agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg
     4141 aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact
     4201 gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg
     4261 gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg
     4321 acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg
     4381 tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg
     4441 ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac
     4501 tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg
     4561 gcaagatcgc cgtgtagtct agataattct accgggtagg ggaggcgctt ttcccaaggc
     4621 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     4681 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     4741 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     4801 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     4861 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     4921 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     4981 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     5041 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
     5101 gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     5161 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     5221 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5281 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5341 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     5401 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5461 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     5521 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     5581 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     5641 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     5701 gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat
     5761 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5821 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5881 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5941 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     6001 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     6061 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     6121 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6181 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6241 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     6301 gggccgcctc cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat
     6361 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     6421 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     6481 agctgtagat cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc
     6541 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     6601 ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta
     6661 caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag
     6721 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg
     6781 gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt
     6841 caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag
     6901 gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc
     6961 tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     7021 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     7081 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     7141 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     7201 agaaatgaat atcagagagt gagaggcctt gacattgcta gcgttttacc gtcgacctct
     7261 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     7321 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     7381 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     7441 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     7501 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     7561 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     7621 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7861 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     8041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     8101 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     8161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     8221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     8281 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     8341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     8401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     8461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8761 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     9001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     9061 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     9121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     9181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     9241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     9301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     9361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     9421 cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa
     9481 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     9541 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     9601 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
     9661 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
     9721 aaattgtatt tgttaaatat gtactacaaa cttagtagtt tttaaagaaa ttgtatttgt
     9781 taaatatgta ctacaaactt agtagt
//
