LOCUS       V014264                 8531 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014264
VERSION     V014264
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8531)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8531
                     /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             2375..2440
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2484..2695
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2708..2976
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3032..3697
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3767..3820
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3827..4618
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    4628..5216
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5288..5521
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5593..5727
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5733..5868
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5906..5922)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5930..5946)
                     /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(5954..5984)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5999..6020)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6308..6896)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7070..7927)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7928..8032)
                     /label="AmpR promoter"
     primer_bind     8506..8522
                     /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 gaattcgcca ccatgctgct atccgtgccg ttgctgctcg gcctcctcgg
     2341 cctggccgtc gccgaccggt cgcacaccgg atccgactac aaagaccatg acggtgatta
     2401 taaagatcat gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg
     2461 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2521 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     2581 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     2641 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     2701 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     2761 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     2821 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     2881 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     2941 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct
     3001 agacgccacc atggagagcg acgagagcgg cctgcccgcc atggagatcg agtgccgcat
     3061 caccggcacc ctgaacggcg tggagttcga gctggtgggc ggcggagagg gcacccccaa
     3121 gcagggccgc atgaccaaca agatgaagag caccaaaggc gccctgacct tcagccccta
     3181 cctgctgagc cacgtgatgg gctacggctt ctaccacttc ggcacctacc ccagcggcta
     3241 cgagaacccc ttcctgcacg ccatcaacaa cggcggctac accaacaccc gcatcgagaa
     3301 gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg ccggccgcgt
     3361 gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga tcttcaccga
     3421 caagatcatc cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg ataacgtgct
     3481 ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc ggctactaca gcttcgtggt
     3541 ggacagccac atgcacttca agagcgccat ccaccccagc atcctgcaga acgggggccc
     3601 catgttcgcc ttccgccgcg tggaggagct gcacagcaac accgagctgg gcatcgtgga
     3661 gtaccagcac gccttcaaga cccccatcgc cttcgccaga tcccgcgctc agtcgtccaa
     3721 ttctgccgtg gacggcaccg ccggacccgg ctccaccgga tctcgcgagg gcagaggaag
     3781 tcttctaaca tgcggtgacg tggaggagaa tcccggccct gccaccatgg ctgaacaaga
     3841 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3901 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3961 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4021 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4081 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4141 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4201 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4261 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4321 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4381 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4441 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4501 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4561 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4621 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4681 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4741 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4801 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4861 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4921 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4981 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5041 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5101 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5161 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcggta
     5221 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     5281 gggggactgg aagggctaat tcactcccaa cgaagataag atctgctttt tgcttgtact
     5341 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5401 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5461 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5521 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     5581 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5641 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5701 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     5761 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5821 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     5881 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5941 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6001 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6061 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6121 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6721 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6841 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6901 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6961 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7021 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7081 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7141 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7201 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7261 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7321 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7381 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7441 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7501 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7561 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7621 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7681 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7741 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7801 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7861 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7921 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7981 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8041 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8101 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8161 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8221 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     8281 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     8341 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     8401 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     8461 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     8521 gtgccaagct g
//
