LOCUS       V015049                 8995 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015049
VERSION     V015049
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8995)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8995
                     /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             2320..3021
                     /label="TEV protease"
                     /note="tobacco etch virus protease (Kapust et al., 2001)"
     CDS             3028..3093
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        3137..3348
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             3361..3629
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3685..4350
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             4420..4473
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             4474..5070
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5074..5662
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5736..5969
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6041..6175
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      6202..6337
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(6370..6386)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6394..6410)
                     /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(6418..6448)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6463..6484)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6772..7360)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7534..8391)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8392..8496)
                     /label="AmpR promoter"
     primer_bind     8970..8986
                     /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 gccaccatgg gcgagagcct tttcaagggc
     2341 ccgagggact acaacccgat ctccagcacc atctgtcacc tgaccaacga gagcgacggt
     2401 cacaccacta gtctgtacgg catcggcttc ggccccttca tcatcaccaa caagcatctg
     2461 ttcaggagga ataacggcac actgctggtg caaagcctgc acggcgtgtt caaagtgaag
     2521 aacacaacca ccctgcaaca gcacctgatc gacggcaggg acatgattat catcaggatg
     2581 cccaaggact tccccccctt tccccagaaa ctgaagttca gggagccaca aagggaggag
     2641 cgaatctgcc tggtgaccac caacttccag accaagtcca tgagcagcat ggtctctgat
     2701 accagctgca ccttccccag cagcgacggc atcttctgga agcactggat tcagacgaag
     2761 gatggccaat gcggcagccc attggtgagc actagggacg gcttcatcgt gggcatccac
     2821 agcgccagca attttaccaa taccaacaac tacttcacga gcgtgccgaa aaacttcatg
     2881 gagctgttga ccaatcaaga ggcgcagcag tgggtgagcg gctggaggct gaacgccgac
     2941 agcgttcttt ggggcggaca taaggtgttc atggtcaagc ccgaggaacc cttccagccc
     3001 gttaaggaag ccactcagct tggatccgac tacaaagacc atgacggtga ttataaagat
     3061 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc
     3121 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     3181 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     3241 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     3301 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     3361 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     3421 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     3481 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     3541 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     3601 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagacgcc
     3661 accatggaga gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc
     3721 accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc caagcagggc
     3781 cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg
     3841 agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac
     3901 cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag
     3961 gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc
     4021 gacttcaagg tggtgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc
     4081 atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacgt gctggtgggc
     4141 agcttcgccc gcaccttcag cctgcgcgac ggcggctact acagcttcgt ggtggacagc
     4201 cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc
     4261 gccttccgcc gcgtggagga gctgcacagc aacaccgagc tgggcatcgt ggagtaccag
     4321 cacgccttca agacccccat cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc
     4381 gtggacggca ccgccggacc cggctccacc ggatctcgcg agggcagagg aagtcttcta
     4441 acatgcggtg acgtggagga gaatcccggc cctatgaccg agtacaagcc cacggtgcgc
     4501 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4561 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4621 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4681 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4741 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4801 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4861 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4921 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4981 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5041 acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta caaaatttgt
     5101 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5161 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5221 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5281 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5341 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5401 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     5461 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     5521 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     5581 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     5641 tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt
     5701 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     5761 aagataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     5821 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5881 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5941 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     6001 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     6061 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6121 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     6181 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6241 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6301 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agaccaaatt
     6361 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6421 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6601 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6661 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6721 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6781 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6841 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6901 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6961 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7021 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7081 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7141 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7201 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7261 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7321 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7381 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7441 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7501 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7561 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7621 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7681 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7741 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7801 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7861 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7921 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7981 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8041 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8101 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8161 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8221 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8281 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8341 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8401 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8461 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8521 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     8581 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     8641 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     8701 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     8761 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     8821 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     8881 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     8941 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
