LOCUS       V014634                 8312 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014634
VERSION     V014634
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8312)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8312
                     /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             2969..3667
                     /label="TagBFP"
                     /note="monomeric blue fluorescent protein"
     CDS             3791..4387
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4391..4979
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5053..5286
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5358..5492
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5519..5654
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5687..5703)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5711..5727)
                     /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(5735..5765)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5780..5801)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6089..6677)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6851..7708)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7709..7813)
                     /label="AmpR promoter"
     primer_bind     8287..8303
                     /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 ccgccaccat gagcgagctg attaaggaga acatgcacat
     3001 gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga
     3061 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
     3121 ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa
     3181 ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga
     3241 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     3301 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc
     3361 tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga
     3421 cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat
     3481 cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg
     3541 cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt
     3601 cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa
     3661 gcttaataga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg
     3721 ctccaccgga tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     3781 tcccggccct atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     3841 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     3901 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     3961 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     4021 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     4081 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     4141 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4201 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4261 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4321 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4381 cggtgcctga aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4441 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     4501 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     4561 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     4621 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     4681 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     4741 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     4801 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     4861 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     4921 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     4981 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5041 aaaagggggg actggaaggg ctaattcact cccaacgaag ataagatctg ctttttgctt
     5101 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5161 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5221 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5281 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     5341 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5401 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5461 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     5521 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5581 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5641 cttttttgga ggcctagact tttgcagaga ccaaattcgt aatcatgtca tagctgtttc
     5701 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5761 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5821 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5881 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7801 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7861 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     7921 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     7981 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     8041 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     8101 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     8161 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     8221 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     8281 gacgttgtaa aacgacggcc agtgccaagc tg
//
