LOCUS       V014254                 8277 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014254
VERSION     V014254
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8277)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             234..414
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     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"
     promoter        2011..2214
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2327..2392
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2436..2647
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2660..2928
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2984..3649
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3719..3772
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3773..4369
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4373..4960
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5034..5267
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5339..5473
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5479..5614
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5652..5668)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5676..5692)
                     /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(5700..5730)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5745..5766)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6054..6642)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6816..7673)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7674..7778)
                     /label="AmpR promoter"
     primer_bind     8252..8268
                     /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 agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gtttagaatc ggatccgact acaaagacca
     2341 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc
     2401 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     2461 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     2521 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     2581 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     2641 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     2701 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     2761 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     2821 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     2881 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     2941 ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat
     3001 cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga
     3061 gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac
     3121 cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta
     3181 ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac
     3241 ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga
     3301 ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt
     3361 gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg
     3421 cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta
     3481 cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca
     3541 gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct
     3601 gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc
     3661 tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga
     3721 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga
     3781 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     3841 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     3901 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     3961 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4021 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4081 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4141 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4201 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4261 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4321 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc
     4381 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4441 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4501 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4561 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4621 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4681 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4741 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4801 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4861 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc tccgccttcg
     4921 ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt taagaccaat
     4981 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5041 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt
     5101 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5161 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5221 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5281 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     5341 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5401 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5461 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     5521 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5581 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat
     5641 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5701 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5761 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5821 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5881 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5941 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6001 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6061 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6121 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6181 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6241 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6301 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6361 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6421 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6481 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6541 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6601 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6661 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6721 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6781 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6841 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6901 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6961 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7021 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7081 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7141 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7201 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7261 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7321 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7381 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7441 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7501 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7561 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7621 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7681 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7741 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7801 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7861 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     7921 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     7981 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     8041 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     8101 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     8161 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     8221 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
