LOCUS       .                      11118 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label="HIV-1 Ψ"
                     /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
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2027..2144
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     polyA_signal    2187..2321
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             complement(2499..2564)
                     /label="3xFLAG"
                     /note="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        complement(2597..2964)
                     /label="TRE3GS promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On® 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription
                     factors"
     promoter        2981..3491
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3510..4253
                     /label="Tet-On® 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        4267..4596
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             4655..6304
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             6374..6427
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             6428..7024
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    7041..7629
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7836..8469
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8646..8676)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8691..8712)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9000..9585)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9759..10616)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(10617..10721)
                     /label="AmpR promoter"
     polyA_signal    10769..10903
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
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 tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgacttaact tgtttattgc agcttataat ggttacaaat
     2221 aaggcaatag catcacaaat ttcacaaata aggcattttt ttcactgcat tctagttttg
     2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctcaaatccc tcggaagctg
     2341 cgcctgtctt aggttggagt gatacatttt tatcactttt acccgtcttt ggattaggca
     2401 gtagctctga cggccctcct gtcttaggtt agtgaaaaat gtcactctct tacccgtcat
     2461 tggctgtcca gcttagctcg caggggaggt ggtctctact tgtcatcgtc atccttgtag
     2521 tcgatgtcat gatctttata atcaccgtca tggtctttgt agtcggatcc gccggcaccg
     2581 gtgtatacgg gaattcttta cgagggtagg aagtggtacg gaaagttggt ataagacaaa
     2641 agtgttgtgg aattgaagtt tactcaaaaa atcagcactc ttttataggc gccctggttt
     2701 acataagcaa agcttatacg ttctctatca ctgataggga gtaaactgga tatacgttct
     2761 ctatcactga tagggagtaa actgtagata cgttctctat cactgatagg gagtaaactg
     2821 gtcatacgtt ctctatcact gatagggagt aaactcctta tacgttctct atcactgata
     2881 gggagtaaag tctgcatacg ttctctatca ctgataggga gtaaactctt catacgttct
     2941 ctatcactga tagggagtaa actcgaggtg ataattccac ggggttgggg ttgcgccttt
     3001 tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac
     3061 gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga
     3121 tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg
     3181 aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta
     3241 gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg
     3301 tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc
     3361 gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat
     3421 tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc
     3481 tctctcccca gggggatcat cgaattacca tgtctagact ggacaagagc aaagtcataa
     3541 actctgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca aggaaactcg
     3601 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     3661 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccactcc tgccccctgg
     3721 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc
     3781 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     3841 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     3901 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     3961 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     4021 aacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     4081 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg
     4141 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     4201 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaaacgc
     4261 gcgaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4321 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4381 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4441 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4501 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4561 tgaggaggct tttttggagg cctaggcttt tgcaaacaag ctgtgaccgg cgcctacgct
     4621 agacgccacc atggagagcg acgagagcgg cctgatggaa gacgccaaaa acataaagaa
     4681 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
     4741 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
     4801 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
     4861 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
     4921 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
     4981 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
     5041 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
     5101 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
     5161 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
     5221 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
     5281 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
     5341 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     5401 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     5461 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
     5521 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     5581 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     5641 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat
     5701 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     5761 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga
     5821 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     5881 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     5941 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt
     6001 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt
     6061 cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     6121 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     6181 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     6241 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc
     6301 cgtgagatcc cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc
     6361 caccggatct cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     6421 cggccctatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     6481 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     6541 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     6601 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     6661 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     6721 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     6781 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     6841 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     6901 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     6961 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     7021 tgcctgaacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg
     7081 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     7141 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     7201 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     7261 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     7321 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     7381 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     7441 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     7501 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     7561 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     7621 cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag
     7681 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     7741 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     7801 agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg
     7861 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca
     7921 gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct
     7981 agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt
     8041 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt
     8101 tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa
     8161 ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg
     8221 cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg
     8281 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     8341 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     8401 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     8461 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     8521 gaatatcaga gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag
     8581 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     8641 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     8701 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     8761 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     8821 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     8881 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     8941 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     9001 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     9061 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     9121 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     9181 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     9241 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     9301 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     9361 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     9421 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     9481 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt
     9541 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     9601 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     9661 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     9721 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     9781 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     9841 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     9901 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     9961 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    10021 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    10081 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    10141 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    10201 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    10261 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    10321 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    10381 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    10441 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    10501 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    10561 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    10621 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    10681 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    10741 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa
    10801 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
    10861 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
    10921 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctagt
    10981 ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta
    11041 tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg
    11101 tactacaaac ttagtagt
//
