LOCUS       V014962                 8454 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014962
VERSION     V014962
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8454)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8454
                     /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             2318..2362
                     /label="S-Tag"
                     /note="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
     CDS             2396..2419
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2423..2446
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2456..2569
                     /label="SBP"
                     /note="streptavidin-binding peptide"
     promoter        2613..2824
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2837..3105
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3161..3826
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3896..3949
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3950..4546
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4550..5137
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5211..5444
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5516..5650
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5656..5791
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5829..5845)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5853..5869)
                     /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(5877..5907)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5922..5943)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6231..6819)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6993..7850)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7851..7955)
                     /label="AmpR promoter"
     primer_bind     8429..8445
                     /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 gaattcgata tccagctcga gggatccaaa gaaaccgctg ctgctaaatt
     2341 cgaacgccag cacatggaca gcggagccgg cgcaggagcc ggcgctgacg cgcctgacta
     2401 caaagacgat gacgacaagg gagattacaa ggatgacgat gacaaaggcg cgtcgatgga
     2461 cgagaagacc accggctggc ggggcggcca cgtggtggag ggcctggccg gcgagctgga
     2521 gcagctgcgg gccaggctgg agcaccaccc tcagggccag cgggagccct agcggccgcg
     2581 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     2641 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
     2701 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     2761 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     2821 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
     2881 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
     2941 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
     3001 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
     3061 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
     3121 gcctacgcta gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga
     3181 gtgccgcatc accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg
     3241 cacccccaag cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt
     3301 cagcccctac ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc
     3361 cagcggctac gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg
     3421 catcgagaag tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc
     3481 cggccgcgtg atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat
     3541 cttcaccgac aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga
     3601 taacgtgctg gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag
     3661 cttcgtggtg gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa
     3721 cgggggcccc atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg
     3781 catcgtggag taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca
     3841 gtcgtccaat tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg
     3901 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta
     3961 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4021 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4081 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4141 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4201 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4261 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4321 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4381 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4441 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     4501 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct
     4561 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4621 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4681 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4741 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4801 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4861 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4921 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     4981 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5041 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc
     5101 tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac
     5161 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5221 aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga
     5281 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     5341 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5401 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     5461 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     5521 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5581 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5641 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5701 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5761 cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg
     5821 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5881 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5941 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6001 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6061 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6121 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6661 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6781 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6841 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6901 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6961 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7021 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7081 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7141 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7201 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7261 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7321 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7381 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7441 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7501 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7561 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7621 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7681 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7741 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7801 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7861 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7981 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8041 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8101 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8161 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8221 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     8281 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8341 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8401 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
