LOCUS       Exported                8647 bp DNA     circular SYN 04-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pRECas1-p15a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8647)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 4, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8647
                     /label=source
                     /organism="synthetic DNA construct"
     terminator      3..56
                     /label=terminator
     primer_bind     57..73
                     /label=M13 rev
     CDS             complement(112..4218)
                     /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    4219..4471
                     /label=Pfdx-E
     misc_feature    4478..4746
                     /label=p1339 (Cac araE)
     misc_feature    4753..4755
                     /label=sgRNA insertion site
     misc_feature    4762..4764
                     /label=HA insertion site
     misc_feature    4767..6391
                     /label=pCB102
     gene            6480..7103
                     /label=catP
     rep_origin      7201..7746
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     oriT            7994..8103
                     /note="incP origin of transfer"
     CDS             8136..8507
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 ggataaaaaa attgtagata aattttataa aatagtttta tctacaattt ttttatcagg
       61 aaacagctat gaccgcggcc gctgtatcca tatgaccatg attacgaatt ctcagtcacc
      121 tcctagctga ctcaaatcaa tgcgtgtttc ataaagacca gtgatggatt gatggataag
      181 agtggcatct aaaacttctt ttgtagacgt atatcgttta cgatcaattg ttgtatcaaa
      241 atatttaaaa gcagcgggag ctccaagatt cgtcaacgta aataaatgaa taatattttc
      301 tgcttgttca cgtattggtt tgtctctatg tttgttatat gcactaagaa ctttatctaa
      361 attggcatct gctaaaataa cacgcttaga aaattcactg atttgctcaa taatctcatc
      421 taaataatgc ttatgctgct ccacaaacaa ttgtttttgt tcgttatctt ctggactacc
      481 cttcaacttt tcataatgac tagctaaata taaaaaattc acatatttgc ttggcagagc
      541 cagctcattt cctttttgta attctccggc actagccagc atccgtttac gaccgttttc
      601 taactcaaaa agactatatt taggtagttt aatgattaag tcttttttaa cttccttata
      661 tcctttagct tctaaaaagt caatcggatt tttttcaaag gaacttcttt ccataattgt
      721 gatccctagt aactctttaa cggattttaa cttcttcgat ttcccttttt ccaccttagc
      781 aaccactagg actgaataag ctaccgttgg actatcaaaa ccaccatatt tttttggatc
      841 ccagtctttt ttacgagcaa taagcttgtc cgaatttctt tttggtaaaa ttgactcctt
      901 ggagaatccg cctgtctgta cttctgtttt cttgacaata ttgacttggg gcatggacaa
      961 tactttgcgc actgtggcaa aatctcgccc tttatcccag acaatttctc cagtttcccc
     1021 attagtttcg attagagggc gtttgcgaat ctctccattt gcaagtgtaa tttctgtttt
     1081 gaagaagttc atgatattag agtaaaagaa atattttgcg gttgctttgc ctatttcttg
     1141 ctcagactta gcaatcattt tacgaacatc ataaacttta taatcaccat agacaaactc
     1201 cgattcaagt tttggatatt tcttaatcaa agcagttcca acgacggcat ttagatacgc
     1261 atcatgggca tgatggtaat tgttaatctc acgtacttta tagaattgga aatcttttcg
     1321 gaagtcagaa actaatttag attttaaggt aatcacttta acctctcgaa taagtttatc
     1381 attttcatcg tatttagtat tcatgcgact atccaaaatt tgtgccacat gcttagtgat
     1441 ttggcgagtt tcaaccaatt ggcgtttgat aaaaccagct ttatcaagtt cactcaaacc
     1501 tccacgttca gctttcgtta aattatcaaa cttacgttga gtgattaact tggcgtttag
     1561 aagttgtctc caatagtttt tcatcttttt gactacttct tcacttggaa cgttatccga
     1621 tttaccacga tttttatcag aacgcgttaa gaccttattg tctattgaat cgtctttaag
     1681 gaaactttgt ggaacaatgt gatcgacatc ataatcactt aaacgattaa tatctaattc
     1741 ttggtccaca tacatgtctc ttccattttg gagataatag agatagagct tttcattttg
     1801 caattgagta ttttcaacag gatgctcttt aagaatctga cttcctaatt ctttgatacc
     1861 ttcttcgatt cgtttcatac gctctcgcga atttttctgg cccttttgag ttgtctgatt
     1921 ttcacgtgcc atttcaataa cgatattttc tggcttatgc cgccccatta ctttgaccaa
     1981 ttcatcaaca acttttacag tctgtaaaat acctttttta atagcagggc taccagctaa
     2041 atttgcaata tgttcatgta aactatcgcc ttgtccagac acttgtgctt tttgaatgtc
     2101 ttctttaaat gtcaaactat catcatggat cagctgcata aaattgcgat tggcaaaacc
     2161 atctgatttc aaaaaatcta atattgtttt gccagattgc ttatccctaa taccattaat
     2221 caattttcga gacaaacgtc cccaaccagt ataacggcga cgtttaagct gtttcatcac
     2281 cttatcatca aagaggtgag catatgtttt aagtctttcc tcaatcatct ccctatcttc
     2341 aaataaggtc aatgttaaaa caatatcctc taagatatct tcattttctt cattatccaa
     2401 aaaatcttta tctttaataa tttttagcaa atcatggtag gtacctaatg aagcattaaa
     2461 tctatcttca actcctgaaa tttcaacact atcaaaacat tctatttttt tgaaataatc
     2521 ttcttttaat tgcttaacgg ttacttttcg atttgttttg aagagtaaat caacaatggc
     2581 tttcttctgt tcacctgaaa gaaatgctgg ttttcgcatt ccttcagtaa catatttgac
     2641 ctttgtcaat tcgttataaa ccgtaaaata ctcataaagc aaactatgtt ttggtagtac
     2701 tttttcattt ggaagatttt tatcaaagtt tgtcatgcgt tcaataaatg attgagctga
     2761 agcaccttta tcgacaactt cttcaaaatt ccatggggta attgtttctt cagacttccg
     2821 agtcatccat gcaaaacgac tattgccacg cgccaatgga ccaacataat aaggaattcg
     2881 aaaagtcaag attttttcaa tcttctcacg attgtctttt aaaaatggat aaaagtcttc
     2941 ttgtcttctc aaaatagcat gcagctcacc caagtgaatt tgatggggaa tagagccgtt
     3001 gtcaaaggtc cgttgcttgc gcagcaaatc ttcacgattt agtttcacca ataattcctc
     3061 agtaccatcc attttttcta aaattggttt gataaattta taaaattctt cttggctagc
     3121 tcccccatca atataacctg catatccgtt ttttgattga tcaaaaaaga tttctttata
     3181 cttttctgga agttgttgtc gaactaaagc ttttaaaaga gtcaagtctt gatgatgttc
     3241 atcgtagcgt ttaatcattg aagctgatag gggagcctta gttatttcag tatttactct
     3301 taggatatct gaaagtaaaa tagcatctga taaattctta gctgccaaaa acaaatcagc
     3361 atattgatct ccaatttgcg ccaataaatt atctaaatca tcatcgtaag tatcttttga
     3421 aagctgtaat ttagcatctt ctgccaaatc aaaatttgat ttaaaattag gggtcaaacc
     3481 caatgacaaa gcaatgagat tcccaaataa gccatttttc ttctcaccgg ggagctgagc
     3541 aatgagattt tctaatcgtc ttgatttact caatcgtgca gaaagaatcg ctttagcatc
     3601 tactccactt gcgttaatag ggttttcttc aaataattga ttgtaggttt gtaccaactg
     3661 gataaatagt ttgtccacat cactattatc aggatttaaa tctccctcaa tcaaaaaatg
     3721 accacgaaac ttaatcatat gcgctaaggc caaatagatt aagcgcaaat ccgctttatc
     3781 agtagaatct accaattttt ttcgcagatg atagatagtt ggatatttct catgataagc
     3841 aacttcatct actatatttc caaaaatagg atgacgttca tgcttcttgt cttcttccac
     3901 caaaaaagac tcttcaagtc gatgaaagaa actatcatct actttcgcca tctcatttga
     3961 aaaaatctcc tgtagataac aaatacgatt cttccgacgt gtataccttc tacgagctgt
     4021 ccgtttgaga cgagtcgctt ccgctgtctc tccactgtca aataaaagag cccctataag
     4081 attttttttg atactgtggc ggtctgtatt tcccagaacc ttgaactttt tagacggaac
     4141 cttatattca tcagtgatca ccgcccatcc gacgctattt gtgccgatat ctaagcctat
     4201 tgagtatttc ttatccatct tgttgttacc tccttagcag ggtgctgcca agggcatcaa
     4261 gacgatgctg gtatcaccgg aacctatagt gagtcgtatt ggtaccgatt tatattttac
     4321 catgttttta ttaattttta aagtaatttt tatataactg tataacctat attttctatg
     4381 gattctatta tcctatctac atctacaaaa taatcatcat atataacact tacttctttt
     4441 ttttccttac ttatttcaca ggctactaca ctctagattt atatttagtc ccttgccttg
     4501 cctacaaggg atttcctatt cctttcattt acaattcata cgtataaaat ccaaattttt
     4561 cttgacattt atacacataa atattatgat ttatataggt aatcgctttc ataaaatata
     4621 ttacccttag gaaatcaaat gattataagt catatatgaa aacgttatat ataattgata
     4681 tgtttacatt tgtaacttag atttctcttt gatttccaca tatataaatc ttaaggagga
     4741 gttttcgtcg acatagacgt caatggcgcg ccgccattat ttttttgaac aattgacaat
     4801 tcatttctta ttttttatta agtgatagtc aaaaggcata acagtgctga atagaaagaa
     4861 atttacagaa aagaaaatta tagaatttag tatgattaat tatactcatt tatgaatgtt
     4921 taattgaata caaaaaaaaa tacttgttat gtattcaatt acgggttaaa atatagacaa
     4981 gttgaaaaat ttaataaaaa aataagtcct cagctcttat atattaagct accaacttag
     5041 tatataagcc aaaacttaaa tgtgctacca acacatcaag ccgttagaga actctatcta
     5101 tagcaatatt tcaaatgtac cgacatacaa gagaaacatt aactatatat attcaattta
     5161 tgagattatc ttaacagata taaatgtaaa ttgcaataag taagatttag aagtttatag
     5221 cctttgtgta ttggaagcag tacgcaaagg cttttttatt tgataaaaat tagaagtata
     5281 tttatttttt cataattaat ttatgaaaat gaaagggggt gagcaaagtg acagaggaaa
     5341 gcagtatctt atcaaataac aaggtattag caatatcatt attgacttta gcagtaaaca
     5401 ttatgacttt tatagtgctt gtagctaagt agtacgaaag ggggagcttt aaaaagctcc
     5461 ttggaataca tagaattcat aaattaattt atgaaaagaa gggcgtatat gaaaacttgt
     5521 aaaaattgca aagagtttat taaagatact gaaatatgca aaatacattc gttgatgatt
     5581 catgataaaa cagtagcaac ctattgcagt aaatacaatg agtcaagatg tttacataaa
     5641 gggaaagtcc aatgtattaa ttgttcaaag atgaaccgat atggatggtg tgccataaaa
     5701 atgagatgtt ttacagagga agaacagaaa aaagaacgta catgcattaa atattatgca
     5761 aggagcttta aaaaagctca tgtaaagaag agtaaaaaga aaaaataatt tatttattaa
     5821 tttaatattg agagtgccga cacagtatgc actaaaaaat atatctgtgg tgtagtgagc
     5881 cgatacaaaa ggatagtcac tcgcattttc ataatacatc ttatgttatg attatgtgtc
     5941 ggtgggactt cacgacgaaa acccacaata aaaaaagagt tcggggtagg gttaagcata
     6001 gttgaggcaa ctaaacaatc aagctaggat atgcagtagc agaccgtaag gtcgttgttt
     6061 aggtgtgttg taatacatac gctattaaga tgtaaaaata cggataccaa tgaagggaaa
     6121 agtataattt ttggatgtag tttgtttgtt catctatggg caaactacgt ccaaagccgt
     6181 ttccaaatct gctaaaaagt atatcctttc taaaatcaaa gtcaagtatg aaatcataaa
     6241 taaagtttaa ttttgaagtt attatgatat tatgtttttc tattaaaata aattaagtat
     6301 atagaatagt ttaataatag tatatactta atgtgataag tgtctgacag tgtcacagaa
     6361 aggatgattg ttatggatta taagcggccg gccagtgggc aagttgaaaa attcacaaaa
     6421 atgtggtata atatctttgt tcattagagc gataaacttg aatttgagag gggacttaga
     6481 tggtatttga aaaaattgat aaaaatagtt ggaacagaaa agagtatttt gaccactact
     6541 ttgcaagtgt accttgtacc tacagcatga ccgttaaagt ggatatcaca caaataaagg
     6601 aaaagggaat gaaactatat cctgcaatgc tttattatat tgcaatgatt gtaaaccgcc
     6661 attcagagtt taggacggca atcaatcaag atggtgaatt ggggatatat gatgagatga
     6721 taccaagcta tacaatattt cacaatgata ctgaaacatt ttccagcctt tggactgagt
     6781 gtaagtctga ctttaaatca tttttagcag attatgaaag tgatacgcaa cggtatggaa
     6841 acaatcatag aatggaagga aagccaaatg ctccggaaaa catttttaat gtatctatga
     6901 taccgtggtc aaccttcgat ggctttaatc tgaatttgca gaaaggatat gattatttga
     6961 ttcctatttt tactatgggg aaatattata aagaagataa caaaattata cttcctttgg
     7021 caattcaagt tcatcacgca gtatgtgacg gatttcacat ttgccgtttt gtaaacgaat
     7081 tgcaggaatt gataaatagt taacttcagg tttgtctgta actaaaaaca agtatttaag
     7141 caaaaacatc gtagaaatac ggtgtttttt gttaccctaa gtttaaacaa gatgatcttc
     7201 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     7261 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
     7321 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
     7381 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
     7441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
     7501 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
     7561 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
     7621 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
     7681 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
     7741 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
     7801 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
     7861 gtagcgagtg ggccccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct
     7921 ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc
     7981 aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct
     8041 tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc
     8101 cggctaccgc cggcgtaaca gatgagggca agcggatggc tgatgaaacc aagccaacca
     8161 ggaagggcag cccacctatc aaggtgtact gccttccaga cgaacgaaga gcgattgagg
     8221 aaaaggcggc ggcggccggc atgagcctgt cggcctacct gctggccgtc ggccagggct
     8281 acaaaatcac gggcgtcgtg gactatgagc acgtccgcga gctggcccgc atcaatggcg
     8341 acctgggccg cctgggcggc ctgctgaaac tctggctcac cgacgacccg cgcacggcgc
     8401 ggttcggtga tgccacgatc ctcgccctgc tggcgaagat cgaagagaag caggacgagc
     8461 ttggcaaggt catgatgggc gtggtccgcc cgagggcaga gccatgactt ttttagccgc
     8521 taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc catcaagaag
     8581 agcgacttcg cggagctggt gaagtacatc accgacgagc aaggcaagac cgatcgggcc
     8641 ccctgca
//
