LOCUS       .                      10643 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"
     promoter        complement(4700..5029)
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             complement(5043..5786)
                     /label="Tet-On® 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        complement(5805..6315)
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     promoter        6332..6699
                     /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             6712..8139
                     /label="tdTomato"
                     /note="tandem dimeric (pseudo-monomeric) derivative of
                     DsRed (Shaner et al., 2004)"
     polyA_signal    complement(8323..8457)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(8500..8617)
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(8689..8730)
                     /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(8879..8923)
                     /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(9108..9341)
                     /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(9838..9963)
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     LTR             complement(10010..10643)
                     /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 tggtcgcgtt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     4741 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     4801 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     4861 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     4921 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     4981 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttcgcgcgtt
     5041 tacccgggga gcatgtcaag gtcaaaatcg tcaagagcgt cagcaggcag catatcaagg
     5101 tcaaagtcgt caagggcatc ggctgggagc atgtctaagt caaaatcgtc aagggcgtcg
     5161 gtcggcccgc cgctttcgca ctttagctgt ttctccaggc cacatatgat tagttccagg
     5221 ccgaaaagga aggcaggttc ggctccctgc cggtcgaaca gctcaattgc ttgtttcaga
     5281 agtgggggca tagaatcggt ggtaggtgtc tctctttcct cttttgctac ttgatgctcc
     5341 tgttcctcca atacgcagcc cagtgtaaag tggcccacgg cggacagagc gtacagtgcg
     5401 ttctccaggg agaagccttg ctgacacagg aacgcgagct gattttccag ggtttcgtac
     5461 tgtttctctg ttgggcgggt gccgagatgc actttagccc cgtcgcgatg tgagaggaga
     5521 gcacagcggt atgacttggc gttgttccgc agaaagtctt gccatgactc gccttccagg
     5581 gggcaggagt gggtatgatg cctgtccagc atctcgattg gcagggcatc gagcagggcc
     5641 cgcttgttct tcacgtgcca gtacagggta ggctgctcaa ctcccagctt ttgagcgagt
     5701 ttccttgtcg tcaggccttc gataccgact ccattgagta attccagagc agagtttatg
     5761 actttgctct tgtccagtct agacatggta attcgatgat ccccctgggg agagaggtcg
     5821 gtgattcggt caacgaggga gccgactgcc gacgtgcgct ccggaggctt gcagaatgcg
     5881 gaacaccgcg cgggcaggaa cagggcccac actaccgccc cacaccccgc ctcccgcacc
     5941 gccccttccc ggccgctgct ctcggcgcgc cctgctgagc agccgctatt ggccacagcc
     6001 catcgcggtc ggcgcgctgc cattgctccc tggcgctgtc cgtctgcgag ggtactagtg
     6061 agacgtgcgg cttccgtttg tcacgtccgg cacgccgcga accgcaagga accttcccga
     6121 cttaggggcg gagcaggaag cgtcgccggg gggcccacaa gggtagcggc gaagatccgg
     6181 gtgacgctgc gaacggacgt gaagaatgtg cgagacccag ggtcggcgcc gctgcgtttc
     6241 ccggaaccac gcccagagca gccgcgtccc tgcgcaaacc cagggctgcc ttggaaaagg
     6301 cgcaacccca accccgtgga attatcacct cgagtttact ccctatcagt gatagagaac
     6361 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca
     6421 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt
     6481 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac
     6541 gtatatccag tttactccct atcagtgata gagaacgtat aagctttgct tatgtaaacc
     6601 agggcgccta taaaagagtg ctgatttttt gagtaaactt caattccaca acacttttgt
     6661 cttataccaa ctttccgtac cacttcctac cctcgtaaag aattcgccac catggtgagc
     6721 aagggcgagg aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg
     6781 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     6841 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     6901 ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac
     6961 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     7021 ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg
     7081 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     7141 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     7201 caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg
     7261 gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc
     7321 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     7381 ctgttcctgg ggcatggcac cggcagcacc ggcagcggca gctccggcac cgcctcctcc
     7441 gaggacaaca acatggccgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     7501 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     7561 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     7621 ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc
     7681 gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     7741 gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac
     7801 aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     7861 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
     7921 atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc
     7981 tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac
     8041 atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc
     8101 caccacctgt tcctgtacgg catggacgag ctgtacaagt agggatccag accacctccc
     8161 ctgcgagcta agctggacag ccaatgacgg gtaagagagt gacatttttc actaacctaa
     8221 gacaggaggg ccgtcagagc tactgcctaa tccaaagacg ggtaaaagtg ataaaaatgt
     8281 atcactccaa cctaagacag gcgcagcttc cgagggattt gagatccaga catgataaga
     8341 tacattgatg agtttggaca aaccaaaact agaatgcagt gaaaaaaatg ccttatttgt
     8401 gaaatttgtg atgctattgc cttatttgta accattataa gctgcaataa acaagttaag
     8461 tcgataaact ggatctctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     8521 ttgtttttgt agttctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     8581 tgcactgtac cccccaatcc ccccttttct tttaaaggcg ataccgtcga gatccgttca
     8641 ctaatcgaat ggatctgtct ctgtctctct ctccaccttc ttcttctatt ccttcgggcc
     8701 tgtcgggtcc cctcggggtt gggaggtggg tctgaaacga taatggtgaa tatccctgcc
     8761 taactctatt cactatagaa agtacagcaa aaactattct taaacctacc aagcctccta
     8821 ctatcattat gaataatttt atataccaca gccaatttgt tatgttaaac caattccaca
     8881 aacttgccca tttatctaat tccaataatt cttgttcatt cttttcttgc tggttttgcg
     8941 attcttcaat taaggagtgt attaagcttg tgtaattgtt aatttctctg tcccactcca
     9001 tccaggtcgt gtgattccaa atctgttcca gagatttatt actccaacta gcattccaag
     9061 gcacagcagt ggtgcaaatg agttttccag agcaacccca aatccccagg agctgttgat
     9121 cctttaggta tctttccaca gccaggattc ttgcctggag ctgcttgatg ccccagactg
     9181 tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag caaattgttc tgctgctgca
     9241 ctataccaga caataattgt ctggcctgta ccgtcagcgt cattgacgct gcgcccatag
     9301 tgcttcctgc tgctcccaag aacccaagga acaaagctcc tattcccact gctctttttt
     9361 ctctctgcac cactcttctc tttgccttgg tgggtgctac tcctaatggt tcaattttta
     9421 ctactttata tttatataat tcacttctcc aattgtccct catatctcct cctccaggtc
     9481 tgaagatcag cggccggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc
     9541 tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag
     9601 ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg
     9661 tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc
     9721 gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc
     9781 ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac
     9841 gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac
     9901 cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga
     9961 gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt
    10021 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
    10081 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
    10141 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca
    10201 aaaagcagct gcttatatgc aggatctgag ggctcgccac tccccagtcc cgcccaggcc
    10261 acgcctccct ggaaagtccc cagcggaaag tcccttgtag caagctcgat atcagcagtt
    10321 cttgaagtac tccggatgca gctctcgggc cacgtgatga aatgctaggc ggctgtcaaa
    10381 cctccactct aacacttctc tctccgggtc atccatccca tgcaggctca cagggtgtaa
    10441 caagctggtg ttctctcctt tattggcctc ttctacctta tctggctcaa ctggtactag
    10501 cttgtagcac catccaaagg tcagtggata tctgacccct ggccctggtg tgtagttctg
    10561 ctaatcaggg aagtagcctt gtgtgtggta gatccacaga tcaaggatat cttgtcttct
    10621 ttgggagtga attagccctt cca
//
