LOCUS       V015022                 9240 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015022
VERSION     V015022
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9240)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9240
                     /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             2297..2362
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2412..2623
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2636..2904
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2963..4612
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             4682..4735
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             4736..5332
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5336..5923
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5997..6230
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6302..6436
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      6442..6577
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(6615..6631)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6639..6655)
                     /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(6663..6693)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6708..6729)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7017..7605)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7779..8636)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8637..8741)
                     /label="AmpR promoter"
     primer_bind     9215..9231
                     /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 gaattcgata
     2281 tcagtctaga ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2341 ctacaaggat gacgatgaca agtaaggatc cgcggccgca aggatctgcg atcgctccgg
     2401 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     2461 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     2521 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     2581 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc
     2641 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag
     2701 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag
     2761 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact
     2821 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt
     2881 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat
     2941 ggagagcgac gagagcggcc tgatggaaga cgccaaaaac ataaagaaag gcccggcgcc
     3001 attctatccg ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata
     3061 cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta
     3121 cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa
     3181 tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt
     3241 gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga
     3301 attgctcaac agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt
     3361 gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga
     3421 ttctaaaacg gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc
     3481 tcccggtttt aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc
     3541 actgatcatg aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag
     3601 aactgcctgc gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc
     3661 ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact
     3721 cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt
     3781 tctgaggagc cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc
     3841 cttcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc
     3901 ttctggtggc gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct
     3961 gccaggtatc aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc
     4021 cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt
     4081 tgtggatctg gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag
     4141 aggtcctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga
     4201 caaggatgga tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt
     4261 catcgttgac cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga
     4321 attggaatcc atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc
     4381 cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat
     4441 gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg
     4501 cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag
     4561 aaaaatcaga gagatcctca taaaggccaa gaagggcgga aagatcgccg tgagatcccg
     4621 cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg
     4681 cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac
     4741 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     4801 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     4861 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4921 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4981 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     5041 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     5101 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     5161 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5221 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5281 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca
     5341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5461 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5521 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5581 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5641 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5701 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     5761 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5821 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct
     5881 tcgccctcag acgagtcgga tctccctttg gccgcctccc cgcctggtac ctttaagacc
     5941 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     6001 agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg ggtctctctg
     6061 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6121 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6181 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     6241 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     6301 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6361 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6421 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg
     6481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6541 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agacggccca
     6601 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6661 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6721 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6841 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6901 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6961 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7021 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7081 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7141 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7201 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7261 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7321 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7381 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7441 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7501 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7561 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7621 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7681 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7741 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7801 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7861 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7921 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7981 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8041 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8101 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8161 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8221 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8281 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8341 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8401 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8461 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8521 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8581 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8641 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8701 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8761 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     8821 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     8881 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     8941 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     9001 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     9061 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     9121 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     9181 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
