LOCUS       .                      10957 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        87..1265
                     /label="EF-1α promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
     CDS             1941..2657
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     misc_feature    2667..3255
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             3327..3560
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3638..3772
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3799..3934
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(3955..3973)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      4141..4596
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4622..4726
                     /label="AmpR promoter"
     CDS             4727..5584
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      5758..6346
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6634..6655
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6670..6700
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     promoter        6769..6787
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6815..7041
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7042..7222
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7269..7394
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7887..8120
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8305..8349
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     promoter        8527..8767
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        10657..10732
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     misc_feature    10788..10905
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
      421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg
     1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
     1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     1861 gcaagcccgg tgccggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
     1921 tcgaagagaa tcctggaccg atggtgtcta agggcgaaga gctgttcacc ggggtggtgc
     1981 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2041 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2101 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2161 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2221 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2281 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2341 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2401 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2461 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2521 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2581 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2641 tggacgagct gtacaagtaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     2701 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2761 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2821 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2881 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2941 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3001 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3061 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3121 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3181 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3241 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
     3301 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     3361 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     3421 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     3481 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     3541 agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt
     3601 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     3661 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3721 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     3781 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3841 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3901 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     3961 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4021 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4081 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     4141 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4201 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4261 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4321 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4381 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4441 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4501 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4561 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4621 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4681 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4741 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4801 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4861 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4921 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4981 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5041 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5101 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5161 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5221 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5281 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5341 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5401 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5461 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5521 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5581 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5641 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5701 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5761 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5821 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5881 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5941 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6001 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6061 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6121 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6181 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6241 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6301 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6361 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6421 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6481 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6541 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6601 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6661 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6721 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     6781 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     6841 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     6901 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     6961 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     7021 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     7081 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7141 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7201 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     7261 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     7321 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     7381 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     7441 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     7501 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     7561 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     7621 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     7681 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     7741 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     7801 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     7861 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     7921 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     7981 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     8041 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     8101 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     8161 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     8221 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     8281 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     8341 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     8401 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     8461 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     8521 cccagagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     8581 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     8641 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     8701 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     8761 aaaggacgaa acaccggaga cggttgtaaa tgagcacaca aaatacacat gctaaaatat
     8821 tatattctat gacctttata aaatcaacca aaatcttctt tttaataact ttagtatcaa
     8881 taattagaat ttttatgttc ctttttgcaa acttttaata aaaatgagca aaataaaaaa
     8941 acgctagttt tagtaactcg cgttgttttc ttcaccttta ataatagcta ctccaccact
     9001 tgttcctaag cggtcagctc ctgcttcaat cattttttga gcatcttcaa atgttctaac
     9061 tccaccagct gctttaacta aagcattgtc tttaacaact gacttcatta gtttaacatc
     9121 ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt ttaacaaatt ctaatccagc
     9181 ttcaacagct atttcacaag ctttcatgat ttcttctttt gttaataaac aattttccat
     9241 aatacattta acaacatgtg atccagctgc tttttttaca gctttcatgt cttctaaaac
     9301 taattcataa tttttgtctt ttaatgcacc aatatttaat accatatcaa tttctgttgc
     9361 accatcttta attgcttcag aaacttcgaa tgcttttgta gctgttgtgc atgcacctag
     9421 aggaaaacct acaacatttg ttattcctac atttgtgcct tttaataatt ctttacaata
     9481 gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca aattcaattg cttcatcaca
     9541 taattgttta atttcagctt tcgtagcatc ttgttttaat aatgtgtgat ctatatattt
     9601 gtttagtttc attttttctc ctatatattc atttttaatt ttaattcttt aataatttcg
     9661 tctactttaa ctttagcgtt ttgaacagat tcaccaacac ctataaaata aatttttagt
     9721 ttaggttcag ttccacttgg gcgaacagca aatcatgact tatcttctaa ataaaatttt
     9781 agtaagtctt gtcctggcat attatacatt ccatcgatgt agtcttcaac attaacaact
     9841 ttaagtccag caatttgagt taagggtgtt gctctcaatg atttcattaa tggttcaatt
     9901 tttaatttct tttcttctgg tttaaaattc aagtttaaag tgaaagtgta atatgcaccc
     9961 atttctttaa ataaatcttc taaatagtct actaatgttt tattttgttt tttataaaat
    10021 caagcagcct ctgctattaa tatagaagct tgtattccat ctttatctct agctgagtca
    10081 tcaattacat atccataact ttcttcataa gcaaaaacaa aatttaatcc gttatcttct
    10141 tctttagcaa tttctctacc cattcattta aatccagtta aagtttttac aatattaact
    10201 ccatattttt catgagcgat tctatcaccc aaatcacttg ttacaaaact tgaatataga
    10261 gccggatttt ttggaatgct atttaagcgt tttagatttg ataattttca atcaattaaa
    10321 attggtcctg tttgatttcc atctaatctt acaaaatgac catcatgttt tattgccatt
    10381 ccaaatctgt cagcatctgg gtcattcata ataataatat ctgcatcatg tttaatacca
    10441 tattcaagcg gtatttttca tgcaggatca aattctggat ttggatttac aacattttta
    10501 aatgtttcat cttcaaatgc atgctcttca acctcaataa cgttatatcc tgattcacgt
    10561 aatatttttg gggtaaattt agttcctgtt ccattaactg cgctaaaaat aatttttaaa
    10621 tcttttttag cttcttgctc ttttttgtac gtctctgttt tagagctaga aatagcaagt
    10681 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttaa
    10741 gcttggcgta actagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa
    10801 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
    10861 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
    10921 gacagcagag atccactttg gcgccggctc gaggggg
//
