LOCUS       V013241                10758 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013241
VERSION     V013241
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10758)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1721..1765
                     /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             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2062..2179
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2231..2534
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2535..2738
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2834..2852
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2882..3006
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3438..3740)
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(4085..4741)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(4742..4844)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(5022..5146)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5154..5870
                     /label="EGFP"
                     /note="enhanced GFP"
     CDS             5880..5903
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             5919..5945
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    5956..6531
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             6532..7128
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    7148..7736
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7811..8044
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        8249..8353
                     /label="AmpR promoter"
     CDS             8354..9211
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      9385..9973
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    10384..10405
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        10420..10450
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    10458..10474
                     /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)."
     primer_bind     10482..10498
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttaattcaca
     2041 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     2101 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     2161 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggactag
     2221 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2281 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2341 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2401 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2461 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2521 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2581 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2641 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2701 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     2761 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccggcgg ccgcagatct
     2821 ctcgaggttg atctaatacg actcactata gggagaccca agctggctag ttaagctatc
     2881 aacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
     2941 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
     3001 actatgaatc aactacttag atggtattag tgacctgtag tcgaccgaca gccttccaaa
     3061 tgttcttcgg gtgatgctgc caacttagtc gaccgacagc cttccaaatg ttcttctcaa
     3121 acggaatcgt cgtatccagc ctactcgcta ttgtcctcaa tgccgtatta aatcataaaa
     3181 agaaataaga aaaagaggtg cgagcctctt ttttgtgtga caaaataaaa acatctacct
     3241 attcatatac gctagtgtca tagtcctgaa aatcatctgc atcaagaaca atttcacaac
     3301 tcttatactt ttctcttaca agtcgttcgg cttcatctgg attttcagcc tctatactta
     3361 ctaaacgtga taaagtttct gtaatttcta ctgtatcgac ctgcagactg gctgtgtata
     3421 agggagcctg acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt
     3481 tcgcggtggc tgagatcagc cacttcttcc ccgataacgg agaccggcac actggccata
     3541 tcggtggtca tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg
     3601 gagactttat ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgcccgggc
     3661 gtgtcaataa tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag
     3721 gtgtaaacct taaactgcat ttcaccagtc cctgttctcg tcagcaaaag agccgttcat
     3781 ttcaataaac cgggcgacct cagccatccc ttcctgattt tccgctttcc agcgttcggc
     3841 acgcagacga cgggcttcat tctgcatggt tgtgcttacc agaccggaga tattgacatc
     3901 atatatgcct tgagcaactg atagctgtcg ctgtcaactg tcactgtaat acgctgcttc
     3961 atagcacacc tctttttgac atacttcggg tatacatatc agtatatatt cttataccgc
     4021 aaaaatcagc gcgcaaatac gcatactgtt atctggcttt tagtaagccg gatccacgcg
     4081 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     4141 catggaagcc atcacagacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     4201 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     4261 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     4321 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     4381 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     4441 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     4501 caccgtcttt cattgccata cggaattccg gatgagcatt catcaggcgg gcaagaatgt
     4561 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     4621 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     4681 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     4741 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     4801 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc
     4861 aaaagttggc ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga
     4921 agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat gctgccaact
     4981 tagtcgacta caggtcacta ataccatcta agtagttgat tcatagtgac tggatatgtt
     5041 gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat
     5101 ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggttcta gacatggtga
     5161 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     5221 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     5281 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     5341 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     5401 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     5461 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     5521 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     5581 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     5641 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     5701 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5761 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5821 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taatctagag
     5881 actacaagga cgacgatgac aagtcggccg ctggaggata cccctacgac gtgcccgact
     5941 acgcctagaa ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     6001 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     6061 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     6121 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     6181 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     6241 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     6301 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     6361 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     6421 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc
     6481 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catgaccgag
     6541 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     6601 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     6661 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     6721 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     6781 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     6841 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6901 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6961 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     7021 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     7081 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg atgcatcgat
     7141 ggatcctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     7201 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     7261 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     7321 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     7381 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     7441 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     7501 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     7561 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     7621 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     7681 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctga
     7741 gatcctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     7801 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     7861 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     7921 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     7981 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     8041 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     8101 tcagagagtg agaggcccgg gttaattaag gaaagggcta gatcattctt gaagacgaaa
     8161 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     8221 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     8281 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     8341 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     8401 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     8461 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     8521 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     8581 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     8641 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     8701 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     8761 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     8821 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     8881 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8941 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     9001 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     9061 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     9121 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     9181 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     9241 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     9301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     9361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     9421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     9481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     9541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     9601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     9661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     9721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     9781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     9841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
    10021 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
    10081 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
    10141 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
    10201 ttaatgcagc aagctcatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
    10261 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
    10321 aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
    10381 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
    10441 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
    10501 gattacgaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    10561 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca
    10621 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact
    10681 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg
    10741 tactacaaac ttagtagt
//
