LOCUS       .                       8287 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     protein_bind    232..253
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        268..298
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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-β-D-thiogalactopyranoside (IPTG)."
     primer_bind     330..346
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        367..385
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        413..639
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    867..992
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1485..1718
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1903..1947
                     /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             2096..2137
                     /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    2214..2331
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2553..2584
                     /label="minP"
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             2646..3362
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     promoter        3459..3816
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3826..4533
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    4543..5131
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5218..5451
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5523..5657
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(5840..5858)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5868..5884)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      6026..6481
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6507..6611
                     /label="AmpR promoter"
     CDS             6612..7469
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      7643..8231
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
      421 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
      481 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta
      541 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
      601 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag
      661 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
      721 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
      781 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
      841 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
      901 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
      961 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     1021 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     1081 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     1141 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     1201 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     1261 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     1321 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     1381 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     1441 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     1501 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     1561 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     1621 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     1681 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     1741 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     1801 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     1861 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     1921 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     1981 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     2041 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     2101 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     2161 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa
     2221 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     2281 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tatcgatcac
     2341 gagactagcc tcctagcccg ggctcgagca gctgaagctt gcatgcctgc agggtaccga
     2401 gctcttacgc gagatcaaag ggggtaagat caaagggggt aagatcaaag gggcgcgaga
     2461 tcaaaggggg taagatcaaa gggggtaaga tcaaaggggg taagatcaaa ggggcgcgcc
     2521 cgcgtgctag cccgggctcg agatctagac tctagagggt atataatgga agctcgaatt
     2581 ccagcttggc attccggtac tgttggtaaa aagcttggca ttccggtact gttggtaaag
     2641 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2701 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2761 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2821 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2881 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2941 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3001 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3061 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3121 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3181 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3241 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3301 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3361 agtaaagcgg ccgcgactct agagtcgaga agcttgatga tctgcgcagc accatggcct
     3421 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     3481 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3541 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3601 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3661 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3721 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3781 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaccatggt gagcaagggc
     3841 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     3901 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     3961 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     4021 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     4081 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     4141 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     4201 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     4261 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     4321 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     4381 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     4441 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     4501 cactccaccg gcggcatgga cgagctgtac aagtaagtcg acaatcaacc tctggattac
     4561 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4621 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4681 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4741 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4801 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4861 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4921 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     4981 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     5041 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     5101 agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta cctttaagac
     5161 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5221 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     5281 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5341 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5401 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     5461 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     5521 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5581 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5641 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     5701 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5761 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5821 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     5881 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     5941 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6001 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6061 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6121 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6181 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6241 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6301 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6361 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6421 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     6481 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6541 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6601 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6661 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6721 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6781 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6841 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6901 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6961 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7021 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7081 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7141 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7201 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7261 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7321 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7381 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7441 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7501 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8281 ttgctca
//
