LOCUS       V014258                 8470 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014258
VERSION     V014258
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8470)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8470
                     /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             2314..2379
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2423..2634
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2647..2915
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2971..3636
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3706..3759
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3766..4557
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    4567..5155
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5227..5460
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5532..5666
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5672..5807
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5845..5861)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5869..5885)
                     /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(5893..5923)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5938..5959)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6247..6835)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7009..7866)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7867..7971)
                     /label="AmpR promoter"
     primer_bind     8445..8461
                     /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 gaattcgaat ttaaatcgga tccgactaca aagaccatga cggtgattat
     2341 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc
     2401 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     2461 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     2521 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     2581 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     2641 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     2701 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     2761 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     2821 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     2881 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     2941 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc
     3001 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag
     3061 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac
     3121 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac
     3181 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag
     3241 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg
     3301 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac
     3361 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg
     3421 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg
     3481 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc
     3541 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag
     3601 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat
     3661 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt
     3721 cttctaacat gcggtgacgt ggaggagaat cccggccctg ccaccatggc tgaacaagat
     3781 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3841 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3901 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3961 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4021 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4081 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4141 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4201 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4261 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4321 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4381 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4441 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4501 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4561 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4621 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4681 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4741 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4801 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4861 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4921 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4981 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     5041 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5101 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcggtac
     5161 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     5221 ggggactgga agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg
     5281 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     5341 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     5401 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     5461 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     5521 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5581 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5641 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     5701 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5761 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     5821 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5881 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5941 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6001 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6061 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6661 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7141 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7861 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7981 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8041 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8101 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8161 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     8221 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     8281 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     8341 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     8401 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     8461 tgccaagctg
//
