LOCUS       .                       8628 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     repeat_region   complement(1..141)
                     /label="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
     promoter        207..447
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        453..528
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        598..1769
                     /label="EF-1α promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
     CDS             1805..4705
                     /label="RfxCas13d"
                     /note="RNA-targeting Type VI CRISPR protein from
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
     CDS             4721..4741
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     misc_feature    4755..5343
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    5375..5851
                     /label="hGH poly(A) signal"
                     /note="human growth hormone polyadenylation signal"
     repeat_region   5891..6031
                     /label="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
     rep_origin      6106..6561
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6843..6947
                     /label="AmpR promoter"
     CDS             6948..7805
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      7979..8567
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgacat tgattattga ctagtgctcc
      181 gaatttctcg aggaattcgg caggaagagg gcctatttcc catgattcct tcatatttgc
      241 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
      301 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
      361 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
      421 tggctttata tatcttgtgg aaaggacgaa acgttttaga gctagaaata gcaagttaaa
      481 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgcta ccgtggggct
      541 gaccgacccc ccgagttcgc tcggggacaa ctagacatac gaaccttttt taagcttggc
      601 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgtgggga
      661 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      721 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
      781 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
      841 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
      901 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
      961 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     1021 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     1081 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     1141 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     1201 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     1261 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     1321 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     1381 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     1441 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     1501 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     1561 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     1621 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     1681 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     1741 gttcaaagtt tttttcttcc atttcaggtc ccgggtaact gatcataatt cgacgctagc
     1801 caccatgatc gaaaaaaaaa agtccttcgc caagggcatg ggcgtgaagt ccacactcgt
     1861 gtccggctcc aaagtgtaca tgacaacctt cgccgaaggc agcgacgcca ggctggaaaa
     1921 gatcgtggag ggcgacagca tcaggagcgt gaatgagggc gaggccttca gcgctgaaat
     1981 ggccgataaa aacgccggct ataagatcgg caacgccaaa ttcagccatc ctaagggcta
     2041 cgccgtggtg gctaacaacc ctctgtatac aggacccgtc cagcaggata tgctcggcct
     2101 gaaggaaact ctggaaaaga ggtacttcgg cgagagcgct gatggcaatg acaatatttg
     2161 tatccaggtg atccataaca tcctggacat tgaaaaaatc ctcgccgaat acattaccaa
     2221 cgccgcctac gccgtcaaca atatctccgg cctggataag gacattattg gattcggcaa
     2281 gttctccaca gtgtatacct acgacgaatt caaagacccc gagcaccata gggccgcttt
     2341 caacaataac gataagctca tcaacgccat caaggcccag tatgacgagt tcgacaactt
     2401 cctcgataac cccagactcg gctatttcgg ccaggccttt ttcagcaagg agggcagaaa
     2461 ttacatcatc aattacggca acgaatgcta tgacattctg gccctcctga gcggactggc
     2521 gcactgggtg gtcgctaaca acgaagaaga gtccaggatc tccaggacct ggctctacaa
     2581 cctcgataag aacctcgaca acgaatacat ctccaccctc aactacctct acgacaggat
     2641 caccaatgag ctgaccaact ccttctccaa gaactccgcc gccaacgtga actatattgc
     2701 cgaaactctg ggaatcaacc ctgccgaatt cgccgaacaa tatttcagat tcagcattat
     2761 gaaagagcag aaaaacctcg gattcaatat caccaagctc agggaagtga tgctggacag
     2821 gaaggatatg tccgagatca ggaaaaatca taaggtgttc gactccatca ggaccaaggt
     2881 ctacaccatg atggactttg tgatttatag gtattacatc gaagaggatg ccaaggtggc
     2941 tgccgccaat aagtccctcc ccgataatga gaagtccctg agcgagaagg atatctttgt
     3001 gattaacctg aggggctcct tcaacgacga ccagaaggat gccctctact acgatgaagc
     3061 taatagaatt tggagaaagc tcgaaaatat catgcacaac atcaaggaat ttaggggaaa
     3121 caagacaaga gagtataaga agaaggacgc ccctagactg cccagaatcc tgcccgctgg
     3181 ccgtgatgtt tccgccttca gcaaactcat gtatgccctg accatgttcc tggatggcaa
     3241 ggagatcaac gacctcctga ccaccctgat taataaattc gataacatcc agagcttcct
     3301 gaaggtgatg cctctcatcg gagtcaacgc taagttcgtg gaggaatacg cctttttcaa
     3361 agactccgcc aagatcgccg atgagctgag gctgatcaag tccttcgcta gaatgggaga
     3421 acctattgcc gatgccagga gggccatgta tatcgacgcc atccgtattt taggaaccaa
     3481 cctgtcctat gatgagctca aggccctcgc cgacaccttt tccctggacg agaacggaaa
     3541 caagctcaag aaaggcaagc acggcatgag aaatttcatt attaataacg tgatcagcaa
     3601 taaaaggttc cactacctga tcagatacgg tgatcctgcc cacctccatg agatcgccaa
     3661 aaacgaggcc gtggtgaagt tcgtgctcgg caggatcgct gacatccaga aaaaacaggg
     3721 ccagaacggc aagaaccaga tcgacaggta ctacgaaact tgtatcggaa aggataaggg
     3781 caagagcgtg agcgaaaagg tggacgctct cacaaagatc atcaccggaa tgaactacga
     3841 ccaattcgac aagaaaagga gcgtcattga ggacaccggc agggaaaacg ccgagaggga
     3901 gaagtttaaa aagatcatca gcctgtacct caccgtgatc taccacatcc tcaagaatat
     3961 tgtcaatatc aacgccaggt acgtcatcgg attccattgc gtcgagcgtg atgctcaact
     4021 gtacaaggag aaaggctacg acatcaatct caagaaactg gaagagaagg gattcagctc
     4081 cgtcaccaag ctctgcgctg gcattgatga aactgccccc gataagagaa aggacgtgga
     4141 aaaggagatg gctgaaagag ccaaggagag cattgacagc ctcgagagcg ccaaccccaa
     4201 gctgtatgcc aattacatca aatacagcga cgagaagaaa gccgaggagt tcaccaggca
     4261 gattaacagg gagaaggcca aaaccgccct gaacgcctac ctgaggaaca ccaagtggaa
     4321 tgtgatcatc agggaggacc tcctgagaat tgacaacaag acatgtaccc tgttcgcaaa
     4381 caaggccgtc gccctggaag tggccaggta tgtccacgcc tatatcaacg acattgccga
     4441 ggtcaattcc tacttccaac tgtaccatta catcatgcag agaattatca tgaatgagag
     4501 gtacgagaaa agcagcggaa aggtgtccga gtacttcgac gctgtgaatg acgagaagaa
     4561 gtacaacgat aggctcctga aactgctgtg tgtgcctttc ggctactgta tccccaggtt
     4621 taagaacctg agcatcgagg ccctgttcga taggaacgag gccgccaagt tcgacaagga
     4681 gaaaaagaag gtgtccggca attccactag tggatccgga cctaagaaaa agaggaaggt
     4741 gtaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     4801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     4861 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     4921 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     4981 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     5041 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5101 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     5161 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     5221 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     5281 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     5341 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     5401 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     5461 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     5521 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     5581 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     5641 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     5701 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     5761 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     5821 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     5881 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     5941 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     6001 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     6061 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     6121 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6181 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6241 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6301 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     6361 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6421 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     6481 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     6541 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     6601 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6721 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     6781 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8101 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     8161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
