LOCUS       V014695                 9797 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014695
VERSION     V014695
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9797)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    458..583
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1493..1537
                     /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             1686..1727
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    1804..1921
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    complement(2056..2085)
                     /label="NFAT binding site"
                     /note="NFAT binding site from the human interleukin 2
                     (IL-2) gene (Mattila et al., 1990)"
     promoter        2093..2206
                     /label="IL-2 promoter"
                     /note="minimal promoter from the human interleukin-2 gene"
     CDS             2254..3903
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             3979..4695
                     /label="EGFP"
                     /note="enhanced GFP"
     promoter        4728..5226
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             5286..5882
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5892..6480
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6554..6787
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6859..6993
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      6999..7134
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(7172..7188)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7196..7212)
                     /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(7220..7250)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7265..7286)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7574..8162)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8336..9193)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9194..9298)
                     /label="AmpR promoter"
     primer_bind     9772..9788
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtaaggatc tgcgatcgct ccggtgcccg tcagtacgcc ttctgtatga
     1981 aacagttttt cctccacgcc ttctgtatga aacagttttt cctccacgcc ttctgtatga
     2041 aacagttttt cctccacgcc ttctgtatga aacagttttt cctcctcgag gacattttga
     2101 cacccccata atatttttcc agaattaaca gtataaattg catctcttgt tcaagagttc
     2161 cctatcactc tctttaatca ctactcacag taacctcaac tcctgcccaa gcttcaagct
     2221 gtgaccggcg cctactctag agctagcgaa ttcatggaag atgccaaaaa cattaagaag
     2281 ggcccagcgc cattctaccc actcgaagac gggaccgccg gcgagcagct gcacaaagcc
     2341 atgaagcgct acgccctggt gcccggcacc atcgccttta ccgacgcaca tatcgaggtg
     2401 gacattacct acgccgagta cttcgagatg agcgttcggc tggcagaagc tatgaagcgc
     2461 tatgggctga atacaaacca tcggatcgtg gtgtgcagcg agaatagctt gcagttcttc
     2521 atgcccgtgt tgggtgccct gttcatcggt gtggctgtgg ccccagctaa cgacatctac
     2581 aacgagcgcg agctgctgaa cagcatgggc atcagccagc ccaccgtcgt attcgtgagc
     2641 aagaaagggc tgcaaaagat cctcaacgtg caaaagaagc taccgatcat acaaaagatc
     2701 atcatcatgg atagcaagac cgactaccag ggcttccaaa gcatgtacac cttcgtgact
     2761 tcccatttgc cacccggctt caacgagtac gacttcgtgc ccgagagctt cgaccgggac
     2821 aaaaccatcg ccctgatcat gaacagtagt ggcagtaccg gattgcccaa gggcgtagcc
     2881 ctaccgcacc gcaccgcttg tgtccgattc agtcatgccc gcgaccccat cttcggcaac
     2941 cagatcatcc ccgacaccgc tatcctcagc gtggtgccat ttcaccacgg cttcggcatg
     3001 ttcaccacgc tgggctactt gatctgcggc tttcgggtcg tgctcatgta ccgcttcgag
     3061 gaggagctat tcttgcgcag cttgcaagac tataagattc aatctgccct gctggtgccc
     3121 acactattta gcttcttcgc taagagcact ctcatcgaca agtacgacct aagcaacttg
     3181 cacgagatcg ccagcggcgg ggcgccgctc agcaaggagg taggtgaggc cgtggccaaa
     3241 cgcttccacc taccaggcat ccgccagggc tacggcctga cagaaacaac cagcgccatt
     3301 ctgatcaccc ccgaagggga cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc
     3361 gaggctaagg tggtggactt ggacaccggt aagacactgg gtgtgaacca gcgcggcgag
     3421 ctgtgcgtcc gtggccccat gatcatgagc ggctacgtta acaaccccga ggctacaaac
     3481 gctctcatcg acaaggacgg ctggctgcac agcggcgaca tcgcctactg ggacgaggac
     3541 gagcacttct tcatcgtgga ccggctgaag agcctgatca aatacaaggg ctaccaggta
     3601 gccccagccg aactggagag catcctgctg caacacccca acatcttcga cgccggggtc
     3661 gccggcctgc ccgacgacga tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac
     3721 ggtaaaacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc
     3781 aagaagctgc gcggtggtgt tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag
     3841 ttggacgccc gcaagatccg cgagattctc attaaggcca agaagggcgg caagatcgcc
     3901 gtgttagcgg ccgctgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     3961 cccggccctt ccggaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     4021 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     4081 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4141 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     4201 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4261 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     4321 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     4381 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     4441 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     4501 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4561 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     4621 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     4681 gacgagctgt acaaggaatt ctaatctaga acgcgattaa ttctaccggg taggggaggc
     4741 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     4801 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     4861 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     4921 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     4981 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     5041 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     5101 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     5161 attctgcacg cttcgaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
     5221 ctttcgaatt caagggcaga ggaagtcttc taacttgagg tgacgtggag gagaatcccg
     5281 gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     5341 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5401 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5461 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5521 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     5581 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     5641 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5701 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5761 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5821 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5881 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5941 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6001 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6061 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6121 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6181 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6241 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     6301 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6361 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6421 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6481 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6541 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct
     6601 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6661 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6721 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6781 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     6841 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6901 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6961 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     7021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     7081 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     7141 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7201 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7261 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7321 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7381 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7441 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7501 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7561 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7621 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7681 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7741 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7801 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7861 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7921 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7981 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     8041 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8101 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8161 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8221 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8281 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8341 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8401 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8461 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8521 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8581 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8641 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8701 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8761 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8821 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8881 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8941 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9001 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9061 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9121 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9181 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9241 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9301 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     9361 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     9421 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     9481 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     9541 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     9601 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     9661 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     9721 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     9781 cggccagtgc caagctg
//
