LOCUS       V013717                 8259 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013717
VERSION     V013717
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8259)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8259
                     /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    1798..1915
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2005..2208
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2308..2373
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2418..2629
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2642..2910
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2966..3631
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3701..3754
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3755..4351
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4355..4942
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5016..5249
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5321..5455
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5461..5596
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5634..5650)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5658..5674)
                     /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(5682..5712)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5727..5748)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6036..6624)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6798..7655)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7656..7760)
                     /label="AmpR promoter"
     primer_bind     8234..8250
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gaatttaaat cggatccgac tacaaagacc atgacggtga ttataaagat
     2341 catgacatcg actacaagga tgacgatgac aagtgagcgg ccgcgaagga tctgcgatcg
     2401 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     2461 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     2521 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     2581 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     2641 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     2701 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     2761 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     2821 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     2881 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc
     2941 caccatggag agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg
     3001 caccctgaac ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg
     3061 ccgcatgacc aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct
     3121 gagccacgtg atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa
     3181 ccccttcctg cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga
     3241 ggacggcggc gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg
     3301 cgacttcaag gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat
     3361 catccgcagc aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg
     3421 cagcttcgcc cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag
     3481 ccacatgcac ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt
     3541 cgccttccgc cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca
     3601 gcacgccttc aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc
     3661 cgtggacggc accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct
     3721 aacatgcggt gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg
     3781 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3841 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3901 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3961 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4021 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4081 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4141 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4201 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4261 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4321 cacctggtgc atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg
     4381 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4441 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4501 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4561 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4621 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4681 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4741 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     4801 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4861 cctgctgccg gctctgcggc ctcttccgcg tctccgcctt cgccctcaga cgagtcggat
     4921 ctccctttgg ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt
     4981 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     5041 aaaataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     5101 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5161 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5221 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     5281 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     5341 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5401 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     5461 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5521 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5581 ggcttttttg gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag
     5641 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5701 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5761 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5821 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5881 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5941 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6001 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6721 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6781 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6841 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6901 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6961 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7021 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7081 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7141 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7201 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7261 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7321 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7381 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7441 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7501 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7561 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7621 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7681 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7741 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7801 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     7861 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     7921 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     7981 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     8041 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     8101 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     8161 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     8221 cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
