LOCUS       V014249                 9871 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014249
VERSION     V014249
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9871)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9871
                     /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             2317..3966
                     /label="luciferase"
                     /note="firefly luciferase"
     promoter        4030..4241
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             4254..4522
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4578..5243
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             5313..5366
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5367..5963
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5967..6554
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6628..6861
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6933..7067
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      7073..7208
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(7246..7262)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7270..7286)
                     /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(7294..7324)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7339..7360)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7648..8236)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8410..9267)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9268..9372)
                     /label="AmpR promoter"
     primer_bind     9846..9862
                     /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 aagatgccaa aaacattaag
     2341 aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa
     2401 gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag
     2461 gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag
     2521 cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc
     2581 ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc
     2641 tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg
     2701 agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag
     2761 atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg
     2821 acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg
     2881 gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta
     2941 gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc
     3001 aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc
     3061 atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc
     3121 gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg
     3181 cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac
     3241 ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc
     3301 aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc
     3361 attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc
     3421 ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc
     3481 gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca
     3541 aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag
     3601 gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag
     3661 gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg
     3721 gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa
     3781 cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc
     3841 gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc
     3901 aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc
     3961 gccgtgtagg atccctgcga tatccagcgc ggccgcgaag gatctgcgat cgctccggtg
     4021 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     4081 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     4141 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     4201 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     4261 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     4321 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     4381 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     4441 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     4501 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac gccaccatgg
     4561 agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga
     4621 acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag ggccgcatga
     4681 ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg
     4741 tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc
     4801 tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg
     4861 gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca
     4921 aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca
     4981 gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg
     5041 cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac agccacatgc
     5101 acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc
     5161 gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac cagcacgcct
     5221 tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct gccgtggacg
     5281 gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt ctaacatgcg
     5341 gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca
     5401 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5461 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5521 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5581 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     5641 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     5701 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5761 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5821 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5881 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5941 gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat
     6001 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     6061 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     6121 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     6181 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     6241 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     6301 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     6361 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6421 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6481 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctcccttt
     6541 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6601 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6661 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6721 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6781 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6841 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6901 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6961 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7021 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     7081 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     7141 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7201 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     7261 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7321 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7381 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7441 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7501 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7561 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7621 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7681 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7741 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7801 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7861 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7921 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7981 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8041 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     8101 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8161 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8221 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8281 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8341 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8401 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8461 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8521 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8581 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8641 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8701 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8761 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8821 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8881 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8941 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9001 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9061 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9121 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9181 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9241 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9361 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9421 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9481 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9541 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9601 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9661 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9721 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9781 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9841 acgttgtaaa acgacggcca gtgccaagct g
//
