LOCUS       .                       8073 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..241
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        266..341
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        403..614
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     CDS             634..1350
                     /label="ECFP"
                     /note="enhanced CFP"
     misc_feature    1376..1964
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2036..2269
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2301..2525
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2571..2999
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    3142..3275
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3312..3328)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(3336..3352)
                     /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)."
     promoter        complement(3360..3390)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3405..3426)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3714..4302)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4476..5333)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(5334..5438)
                     /label="AmpR promoter"
     enhancer        5704..6083
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6084..6286
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    6528..6653
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7146..7379
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7564..7608
                     /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             7757..7798
                     /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    7906..8023
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gagacggacg tctctgtttt agagctagaa atagcaagtt aaaataaggc
      301 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttcgctagct
      361 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
      421 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
      481 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      541 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      601 ccgccagaac acaggaccgg ttctagagcc accatggtga gcaagggcga ggagctgttc
      661 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      721 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      781 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg
      841 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      901 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      961 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1021 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac
     1081 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1141 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1201 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1261 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1321 atcactctcg gcatggacga gctgtacaag tagctcgagt acgcgttaag tcgacaatca
     1381 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     1441 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     1501 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     1561 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     1621 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     1681 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     1741 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     1801 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     1861 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     1921 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
     1981 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     2041 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     2101 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     2161 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     2221 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
     2281 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2341 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2401 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2461 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2521 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2581 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2641 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2701 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2761 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2821 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2881 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2941 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3001 attctgtgga atcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3061 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3121 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3181 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3241 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3301 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3361 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3421 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3481 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5461 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     5521 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     5581 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     5641 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     5701 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     5761 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5821 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5881 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5941 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6001 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6061 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     6121 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     6181 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     6241 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
     6301 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6361 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6421 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6481 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     6541 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     6601 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     6661 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     6721 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     6781 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     6841 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     6901 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     6961 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     7021 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     7081 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     7141 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     7201 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     7261 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     7321 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     7381 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     7441 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     7501 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     7561 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     7621 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     7681 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     7741 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     7801 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     7861 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     7921 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     7981 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     8041 cagcagagat ccagtttggt taattaaggt acc
//
