LOCUS       .                      10783 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     polyA_signal    complement(216..350)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        398..502
                     /label="AmpR promoter"
     CDS             503..1360
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      1534..2119
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2407..2428
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2443..2473
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     LTR             complement(2650..3283)
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     misc_feature    complement(3490..4078)
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(4095..4691)
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     CDS             complement(4701..4754)
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             complement(4824..5489)
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     promoter        complement(5540..5869)
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             complement(5883..6626)
                     /label="Tet-On® 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        complement(6645..7155)
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     promoter        7172..7539
                     /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"
     CDS             7555..8262
                     /label="TEV protease"
                     /note="tobacco etch virus protease (Kapust et al., 2001)"
     polyA_signal    complement(8463..8597)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(8640..8757)
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(8829..8870)
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     CDS             complement(9019..9063)
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     misc_feature    complement(9248..9481)
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     misc_feature    complement(9978..10103)
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     LTR             complement(10150..10783)
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaaact actaagtttg
       61 tagtacatat ttaacaaata caatttcttt aaaatgaaaa taattcagag gaatcacagg
      121 tttagagtaa atgaaaccac taggtaattg gcagtggtaa tagggtatgg ggtgggaagt
      181 ttgggatgat tttggttagc ttgagttatc cagttgatcc agacatgata agatacattg
      241 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      301 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt gatctcccga
      361 tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1561 ctgctgcttg caaacaaaaa aaccgctacc agcggtggtt tgtttgccgg atcaagagct
     1621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2281 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2341 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2401 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2461 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2521 ccatgattac gccaagctct agctagaggt cgacggtaaa acgctagcaa tgtcaaggcc
     2581 tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata agatgacatg
     2641 aactactact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     2701 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     2761 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     2821 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     2881 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag
     2941 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga
     3001 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca
     3061 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta
     3121 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct
     3181 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga
     3241 tcaaggatat cttgtcttcg ttgggagtga attagccctt ccagtcccct cttttctttt
     3301 aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg tacctgaggt
     3361 gtgactggaa aacccacctc ctcctcctct tgtgcttcta gccaggcaca atcagcattg
     3421 gtagctgctg tattgctact tgtgattgct ccatgttttt ctaggtctcg actgcagaat
     3481 taattccagg cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa
     3541 gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga
     3601 gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga
     3661 gcagccatgg aaaggacgtc agcttccccg acaacaccac ggaattgtca gtgcccaaca
     3721 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag
     3781 ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag
     3841 tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca
     3901 actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac
     3961 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     4021 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt
     4081 tccagacgcg ttcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     4141 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     4201 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     4261 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     4321 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     4381 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     4441 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     4501 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     4561 tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     4621 gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     4681 tactcggtca tggtcgcgtc agggccggga ttctcctcca cgtcaccgca tgttagaaga
     4741 cttcctctgc cctcgcgaga tccggtggag ccgggtccgg cggtgccgtc cacggcagaa
     4801 ttggacgact gagcgcggga tctggcgaag gcgatggggg tcttgaaggc gtgctggtac
     4861 tccacgatgc ccagctcggt gttgctgtgc agctcctcca cgcggcggaa ggcgaacatg
     4921 gggcccccgt tctgcaggat gctggggtgg atggcgctct tgaagtgcat gtggctgtcc
     4981 accacgaagc tgtagtagcc gccgtcgcgc aggctgaagg tgcgggcgaa gctgcccacc
     5041 agcacgttat cgcccatggg gtgcaggtgc tccacggtgg cgttgctgcg gatgatcttg
     5101 tcggtgaaga tcacgctgtc ctcggggaag ccggtgccca ccaccttgaa gtcgccgatc
     5161 acgcggccgg cctcgtagcg gtagctgaag ctcacgtgca gcacgccgcc gtcctcgtac
     5221 ttctcgatgc gggtgttggt gtagccgccg ttgttgatgg cgtgcaggaa ggggttctcg
     5281 tagccgctgg ggtaggtgcc gaagtggtag aagccgtagc ccatcacgtg gctcagcagg
     5341 taggggctga aggtcagggc gcctttggtg ctcttcatct tgttggtcat gcggccctgc
     5401 ttgggggtgc cctctccgcc gcccaccagc tcgaactcca cgccgttcag ggtgccggtg
     5461 atgcggcact cgatctccat ggcgggcagg ccgctctcgt cgctctccat ggtggcgtct
     5521 agcgtaggcg ccggtcacat ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     5581 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     5641 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     5701 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5761 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     5821 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttcgcgcgtt
     5881 tacccgggga gcatgtcaag gtcaaaatcg tcaagagcgt cagcaggcag catatcaagg
     5941 tcaaagtcgt caagggcatc ggctgggagc atgtctaagt caaaatcgtc aagggcgtcg
     6001 gtcggcccgc cgctttcgca ctttagctgt ttctccaggc cacatatgat tagttccagg
     6061 ccgaaaagga aggcaggttc ggctccctgc cggtcgaaca gctcaattgc ttgtttcaga
     6121 agtgggggca tagaatcggt ggtaggtgtc tctctttcct cttttgctac ttgatgctcc
     6181 tgttcctcca atacgcagcc cagtgtaaag tggcccacgg cggacagagc gtacagtgcg
     6241 ttctccaggg agaagccttg ctgacacagg aacgcgagct gattttccag ggtttcgtac
     6301 tgtttctctg ttgggcgggt gccgagatgc actttagccc cgtcgcgatg tgagaggaga
     6361 gcacagcggt atgacttggc gttgttccgc agaaagtctt gccatgactc gccttccagg
     6421 gggcaggagt gggtatgatg cctgtccagc atctcgattg gcagggcatc gagcagggcc
     6481 cgcttgttct tcacgtgcca gtacagggta ggctgctcaa ctcccagctt ttgagcgagt
     6541 ttccttgtcg tcaggccttc gataccgact ccattgagta attccagagc agagtttatg
     6601 actttgctct tgtccagtct agacatggta attcgatgat ccccctgggg agagaggtcg
     6661 gtgattcggt caacgaggga gccgactgcc gacgtgcgct ccggaggctt gcagaatgcg
     6721 gaacaccgcg cgggcaggaa cagggcccac actaccgccc cacaccccgc ctcccgcacc
     6781 gccccttccc ggccgctgct ctcggcgcgc cctgctgagc agccgctatt ggccacagcc
     6841 catcgcggtc ggcgcgctgc cattgctccc tggcgctgtc cgtctgcgag ggtactagtg
     6901 agacgtgcgg cttccgtttg tcacgtccgg cacgccgcga accgcaagga accttcccga
     6961 cttaggggcg gagcaggaag cgtcgccggg gggcccacaa gggtagcggc gaagatccgg
     7021 gtgacgctgc gaacggacgt gaagaatgtg cgagacccag ggtcggcgcc gctgcgtttc
     7081 ccggaaccac gcccagagca gccgcgtccc tgcgcaaacc cagggctgcc ttggaaaagg
     7141 cgcaacccca accccgtgga attatcacct cgagtttact ccctatcagt gatagagaac
     7201 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca
     7261 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt
     7321 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac
     7381 gtatatccag tttactccct atcagtgata gagaacgtat aagctttgct tatgtaaacc
     7441 agggcgccta taaaagagtg ctgatttttt gagtaaactt caattccaca acacttttgt
     7501 cttataccaa ctttccgtac cacttcctac cctcgtaaag aattcgccac catgggcgag
     7561 agcttgttta agggaccacg tgattacaac ccgatatcga gcaccatttg tcatttgacg
     7621 aatgaatctg atgggcacac aacatcgttg tatggtattg gatttggtcc cttcatcatt
     7681 acaaacaagc acttgtttag aagaaataat ggaacactgt tggtccaatc actacatggt
     7741 gtattcaagg tcaagaacac cacgactttg caacaacacc tcattgatgg gagggacatg
     7801 ataattattc gcatgcctaa ggatttccca ccatttcctc aaaagctgaa atttagagag
     7861 ccacaaaggg aagagcgcat atgtcttgtg acaaccaact tccaaactaa gagcatgtct
     7921 agcatggtgt cagacactag ttgcacattc ccttcatctg atggcatatt ctggaagcat
     7981 tggattcaaa ccaaggatgg gcagtgtggc agtccattag tatcaactag agatgggttc
     8041 attgttggta tacactcagc atcgaatttc accaacacaa acaattattt cacaagcgtg
     8101 ccgaaaaact tcatggaatt gttgacaaat caggaggcgc agcagtgggt tagtggttgg
     8161 cgattaaatg ctgactcagt attgtggggg ggccataaag ttttcatgaa caaacctgaa
     8221 gagccttttc agccagttaa ggaagcgact caactcatga atgaattggt gtactcgaac
     8281 taggatccag accacctccc ctgcgagcta agctggacag ccaatgacgg gtaagagagt
     8341 gacatttttc actaacctaa gacaggaggg ccgtcagagc tactgcctaa tccaaagacg
     8401 ggtaaaagtg ataaaaatgt atcactccaa cctaagacag gcgcagcttc cgagggattt
     8461 gagatccaga catgataaga tacattgatg agtttggaca aaccaaaact agaatgcagt
     8521 gaaaaaaatg ccttatttgt gaaatttgtg atgctattgc cttatttgta accattataa
     8581 gctgcaataa acaagttaag tcgataaact ggatctctgc tgtccctgta ataaacccga
     8641 aaattttgaa tttttgtaat ttgtttttgt agttctttag tttgtatgtc tgttgctatt
     8701 atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaggcg
     8761 ataccgtcga gatccgttca ctaatcgaat ggatctgtct ctgtctctct ctccaccttc
     8821 ttcttctatt ccttcgggcc tgtcgggtcc cctcggggtt gggaggtggg tctgaaacga
     8881 taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct
     8941 taaacctacc aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt
     9001 tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt
     9061 cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt
     9121 aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt
     9181 actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca
     9241 aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag
     9301 ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag
     9361 caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt
     9421 cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc
     9481 tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac
     9541 tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc aattgtccct
     9601 catatctcct cctccaggtc tgaagatcag cggccggccg cttgctgtgc ggtggtctta
     9661 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     9721 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     9781 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     9841 aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     9901 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     9961 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
    10021 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
    10081 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
    10141 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
    10201 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
    10261 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
    10321 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
    10381 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag
    10441 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga
    10501 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca
    10561 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta
    10621 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct
    10681 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga
    10741 tcaaggatat cttgtcttct ttgggagtga attagccctt cca
//
