LOCUS       Exported                8842 bp DNA     circular SYN 04-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pRECas1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8842)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 4, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8842
                     /label=source
                     /organism="synthetic DNA construct"
     terminator      140..193
                     /label=terminator
     primer_bind     194..210
                     /label=M13 rev
     CDS             complement(249..4355)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     misc_feature    4356..4608
                     /label=Pfdx-E
     misc_feature    4615..4883
                     /label=p1339 (Cac araE)
     misc_feature    4890..4892
                     /label=sgRNA insertion site
     misc_feature    4899..4901
                     /label=HA insertion site
     misc_feature    4904..6528
                     /label=pCB102
     gene            6617..7240
                     /label=catP
     rep_origin      7417..8005
                     /label=ori
     oriT            8326..8435
                     /label=incP origin of transfer
     CDS             8468..8839
                     /label=traJ
ORIGIN
        1 ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc
       61 catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc aaggcaagac
      121 cgatcgggcc ccctgcagga taaaaaaatt gtagataaat tttataaaat agttttatct
      181 acaatttttt tatcaggaaa cagctatgac cgcggccgct gtatccatat gaccatgatt
      241 acgaattctc agtcacctcc tagctgactc aaatcaatgc gtgtttcata aagaccagtg
      301 atggattgat ggataagagt ggcatctaaa acttcttttg tagacgtata tcgtttacga
      361 tcaattgttg tatcaaaata tttaaaagca gcgggagctc caagattcgt caacgtaaat
      421 aaatgaataa tattttctgc ttgttcacgt attggtttgt ctctatgttt gttatatgca
      481 ctaagaactt tatctaaatt ggcatctgct aaaataacac gcttagaaaa ttcactgatt
      541 tgctcaataa tctcatctaa ataatgctta tgctgctcca caaacaattg tttttgttcg
      601 ttatcttctg gactaccctt caacttttca taatgactag ctaaatataa aaaattcaca
      661 tatttgcttg gcagagccag ctcatttcct ttttgtaatt ctccggcact agccagcatc
      721 cgtttacgac cgttttctaa ctcaaaaaga ctatatttag gtagtttaat gattaagtct
      781 tttttaactt ccttatatcc tttagcttct aaaaagtcaa tcggattttt ttcaaaggaa
      841 cttctttcca taattgtgat ccctagtaac tctttaacgg attttaactt cttcgatttc
      901 cctttttcca ccttagcaac cactaggact gaataagcta ccgttggact atcaaaacca
      961 ccatattttt ttggatccca gtctttttta cgagcaataa gcttgtccga atttcttttt
     1021 ggtaaaattg actccttgga gaatccgcct gtctgtactt ctgttttctt gacaatattg
     1081 acttggggca tggacaatac tttgcgcact gtggcaaaat ctcgcccttt atcccagaca
     1141 atttctccag tttccccatt agtttcgatt agagggcgtt tgcgaatctc tccatttgca
     1201 agtgtaattt ctgttttgaa gaagttcatg atattagagt aaaagaaata ttttgcggtt
     1261 gctttgccta tttcttgctc agacttagca atcattttac gaacatcata aactttataa
     1321 tcaccataga caaactccga ttcaagtttt ggatatttct taatcaaagc agttccaacg
     1381 acggcattta gatacgcatc atgggcatga tggtaattgt taatctcacg tactttatag
     1441 aattggaaat cttttcggaa gtcagaaact aatttagatt ttaaggtaat cactttaacc
     1501 tctcgaataa gtttatcatt ttcatcgtat ttagtattca tgcgactatc caaaatttgt
     1561 gccacatgct tagtgatttg gcgagtttca accaattggc gtttgataaa accagcttta
     1621 tcaagttcac tcaaacctcc acgttcagct ttcgttaaat tatcaaactt acgttgagtg
     1681 attaacttgg cgtttagaag ttgtctccaa tagtttttca tctttttgac tacttcttca
     1741 cttggaacgt tatccgattt accacgattt ttatcagaac gcgttaagac cttattgtct
     1801 attgaatcgt ctttaaggaa actttgtgga acaatgtgat cgacatcata atcacttaaa
     1861 cgattaatat ctaattcttg gtccacatac atgtctcttc cattttggag ataatagaga
     1921 tagagctttt cattttgcaa ttgagtattt tcaacaggat gctctttaag aatctgactt
     1981 cctaattctt tgataccttc ttcgattcgt ttcatacgct ctcgcgaatt tttctggccc
     2041 ttttgagttg tctgattttc acgtgccatt tcaataacga tattttctgg cttatgccgc
     2101 cccattactt tgaccaattc atcaacaact tttacagtct gtaaaatacc ttttttaata
     2161 gcagggctac cagctaaatt tgcaatatgt tcatgtaaac tatcgccttg tccagacact
     2221 tgtgcttttt gaatgtcttc tttaaatgtc aaactatcat catggatcag ctgcataaaa
     2281 ttgcgattgg caaaaccatc tgatttcaaa aaatctaata ttgttttgcc agattgctta
     2341 tccctaatac cattaatcaa ttttcgagac aaacgtcccc aaccagtata acggcgacgt
     2401 ttaagctgtt tcatcacctt atcatcaaag aggtgagcat atgttttaag tctttcctca
     2461 atcatctccc tatcttcaaa taaggtcaat gttaaaacaa tatcctctaa gatatcttca
     2521 ttttcttcat tatccaaaaa atctttatct ttaataattt ttagcaaatc atggtaggta
     2581 cctaatgaag cattaaatct atcttcaact cctgaaattt caacactatc aaaacattct
     2641 atttttttga aataatcttc ttttaattgc ttaacggtta cttttcgatt tgttttgaag
     2701 agtaaatcaa caatggcttt cttctgttca cctgaaagaa atgctggttt tcgcattcct
     2761 tcagtaacat atttgacctt tgtcaattcg ttataaaccg taaaatactc ataaagcaaa
     2821 ctatgttttg gtagtacttt ttcatttgga agatttttat caaagtttgt catgcgttca
     2881 ataaatgatt gagctgaagc acctttatcg acaacttctt caaaattcca tggggtaatt
     2941 gtttcttcag acttccgagt catccatgca aaacgactat tgccacgcgc caatggacca
     3001 acataataag gaattcgaaa agtcaagatt ttttcaatct tctcacgatt gtcttttaaa
     3061 aatggataaa agtcttcttg tcttctcaaa atagcatgca gctcacccaa gtgaatttga
     3121 tggggaatag agccgttgtc aaaggtccgt tgcttgcgca gcaaatcttc acgatttagt
     3181 ttcaccaata attcctcagt accatccatt ttttctaaaa ttggtttgat aaatttataa
     3241 aattcttctt ggctagctcc cccatcaata taacctgcat atccgttttt tgattgatca
     3301 aaaaagattt ctttatactt ttctggaagt tgttgtcgaa ctaaagcttt taaaagagtc
     3361 aagtcttgat gatgttcatc gtagcgttta atcattgaag ctgatagggg agccttagtt
     3421 atttcagtat ttactcttag gatatctgaa agtaaaatag catctgataa attcttagct
     3481 gccaaaaaca aatcagcata ttgatctcca atttgcgcca ataaattatc taaatcatca
     3541 tcgtaagtat cttttgaaag ctgtaattta gcatcttctg ccaaatcaaa atttgattta
     3601 aaattagggg tcaaacccaa tgacaaagca atgagattcc caaataagcc atttttcttc
     3661 tcaccgggga gctgagcaat gagattttct aatcgtcttg atttactcaa tcgtgcagaa
     3721 agaatcgctt tagcatctac tccacttgcg ttaatagggt tttcttcaaa taattgattg
     3781 taggtttgta ccaactggat aaatagtttg tccacatcac tattatcagg atttaaatct
     3841 ccctcaatca aaaaatgacc acgaaactta atcatatgcg ctaaggccaa atagattaag
     3901 cgcaaatccg ctttatcagt agaatctacc aatttttttc gcagatgata gatagttgga
     3961 tatttctcat gataagcaac ttcatctact atatttccaa aaataggatg acgttcatgc
     4021 ttcttgtctt cttccaccaa aaaagactct tcaagtcgat gaaagaaact atcatctact
     4081 ttcgccatct catttgaaaa aatctcctgt agataacaaa tacgattctt ccgacgtgta
     4141 taccttctac gagctgtccg tttgagacga gtcgcttccg ctgtctctcc actgtcaaat
     4201 aaaagagccc ctataagatt ttttttgata ctgtggcggt ctgtatttcc cagaaccttg
     4261 aactttttag acggaacctt atattcatca gtgatcaccg cccatccgac gctatttgtg
     4321 ccgatatcta agcctattga gtatttctta tccatcttgt tgttacctcc ttagcagggt
     4381 gctgccaagg gcatcaagac gatgctggta tcaccggaac ctatagtgag tcgtattggt
     4441 accgatttat attttaccat gtttttatta atttttaaag taatttttat ataactgtat
     4501 aacctatatt ttctatggat tctattatcc tatctacatc tacaaaataa tcatcatata
     4561 taacacttac ttcttttttt tccttactta tttcacaggc tactacactc tagatttata
     4621 tttagtccct tgccttgcct acaagggatt tcctattcct ttcatttaca attcatacgt
     4681 ataaaatcca aatttttctt gacatttata cacataaata ttatgattta tataggtaat
     4741 cgctttcata aaatatatta cccttaggaa atcaaatgat tataagtcat atatgaaaac
     4801 gttatatata attgatatgt ttacatttgt aacttagatt tctctttgat ttccacatat
     4861 ataaatctta aggaggagtt ttcgtcgaca tagacgtcaa tggcgcgccg ccattatttt
     4921 tttgaacaat tgacaattca tttcttattt tttattaagt gatagtcaaa aggcataaca
     4981 gtgctgaata gaaagaaatt tacagaaaag aaaattatag aatttagtat gattaattat
     5041 actcatttat gaatgtttaa ttgaatacaa aaaaaaatac ttgttatgta ttcaattacg
     5101 ggttaaaata tagacaagtt gaaaaattta ataaaaaaat aagtcctcag ctcttatata
     5161 ttaagctacc aacttagtat ataagccaaa acttaaatgt gctaccaaca catcaagccg
     5221 ttagagaact ctatctatag caatatttca aatgtaccga catacaagag aaacattaac
     5281 tatatatatt caatttatga gattatctta acagatataa atgtaaattg caataagtaa
     5341 gatttagaag tttatagcct ttgtgtattg gaagcagtac gcaaaggctt ttttatttga
     5401 taaaaattag aagtatattt attttttcat aattaattta tgaaaatgaa agggggtgag
     5461 caaagtgaca gaggaaagca gtatcttatc aaataacaag gtattagcaa tatcattatt
     5521 gactttagca gtaaacatta tgacttttat agtgcttgta gctaagtagt acgaaagggg
     5581 gagctttaaa aagctccttg gaatacatag aattcataaa ttaatttatg aaaagaaggg
     5641 cgtatatgaa aacttgtaaa aattgcaaag agtttattaa agatactgaa atatgcaaaa
     5701 tacattcgtt gatgattcat gataaaacag tagcaaccta ttgcagtaaa tacaatgagt
     5761 caagatgttt acataaaggg aaagtccaat gtattaattg ttcaaagatg aaccgatatg
     5821 gatggtgtgc cataaaaatg agatgtttta cagaggaaga acagaaaaaa gaacgtacat
     5881 gcattaaata ttatgcaagg agctttaaaa aagctcatgt aaagaagagt aaaaagaaaa
     5941 aataatttat ttattaattt aatattgaga gtgccgacac agtatgcact aaaaaatata
     6001 tctgtggtgt agtgagccga tacaaaagga tagtcactcg cattttcata atacatctta
     6061 tgttatgatt atgtgtcggt gggacttcac gacgaaaacc cacaataaaa aaagagttcg
     6121 gggtagggtt aagcatagtt gaggcaacta aacaatcaag ctaggatatg cagtagcaga
     6181 ccgtaaggtc gttgtttagg tgtgttgtaa tacatacgct attaagatgt aaaaatacgg
     6241 ataccaatga agggaaaagt ataatttttg gatgtagttt gtttgttcat ctatgggcaa
     6301 actacgtcca aagccgtttc caaatctgct aaaaagtata tcctttctaa aatcaaagtc
     6361 aagtatgaaa tcataaataa agtttaattt tgaagttatt atgatattat gtttttctat
     6421 taaaataaat taagtatata gaatagttta ataatagtat atacttaatg tgataagtgt
     6481 ctgacagtgt cacagaaagg atgattgtta tggattataa gcggccggcc agtgggcaag
     6541 ttgaaaaatt cacaaaaatg tggtataata tctttgttca ttagagcgat aaacttgaat
     6601 ttgagagggg acttagatgg tatttgaaaa aattgataaa aatagttgga acagaaaaga
     6661 gtattttgac cactactttg caagtgtacc ttgtacctac agcatgaccg ttaaagtgga
     6721 tatcacacaa ataaaggaaa agggaatgaa actatatcct gcaatgcttt attatattgc
     6781 aatgattgta aaccgccatt cagagtttag gacggcaatc aatcaagatg gtgaattggg
     6841 gatatatgat gagatgatac caagctatac aatatttcac aatgatactg aaacattttc
     6901 cagcctttgg actgagtgta agtctgactt taaatcattt ttagcagatt atgaaagtga
     6961 tacgcaacgg tatggaaaca atcatagaat ggaaggaaag ccaaatgctc cggaaaacat
     7021 ttttaatgta tctatgatac cgtggtcaac cttcgatggc tttaatctga atttgcagaa
     7081 aggatatgat tatttgattc ctatttttac tatggggaaa tattataaag aagataacaa
     7141 aattatactt cctttggcaa ttcaagttca tcacgcagta tgtgacggat ttcacatttg
     7201 ccgttttgta aacgaattgc aggaattgat aaatagttaa cttcaggttt gtctgtaact
     7261 aaaaacaagt atttaagcaa aaacatcgta gaaatacggt gttttttgtt accctaagtt
     7321 taaactcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7381 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7441 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7501 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     8041 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     8101 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     8161 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc agggccccct gcttcggggt
     8221 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     8281 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     8341 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     8401 gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg
     8461 caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta tcaaggtgta
     8521 ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg gcatgagcct
     8581 gtcggcctac ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg tggactatga
     8641 gcacgtccgc gagctggccc gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa
     8701 actctggctc accgacgacc cgcgcacggc gcggttcggt gatgccacga tcctcgccct
     8761 gctggcgaag atcgaagaga agcaggacga gcttggcaag gtcatgatgg gcgtggtccg
     8821 cccgagggca gagccatgac tt
//
