LOCUS       sequence-329657-        9659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes aTc-inducible spCas9 and cumate-inducible MutLE32K for 
            transient suppression of mismatch repair.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9659)
  AUTHORS   Ellington AJ, Reisch CR
  TITLE     Efficient and iterative retron-mediated in vivo recombineering in 
            Escherichia coli.
  JOURNAL   Synth Biol (Oxf). 2022 May 3;7(1):ysac007. doi: 
            10.1093/synbio/ysac007. eCollection 2022.
  PUBMED    35673614
REFERENCE   2  (bases 1 to 9659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synth Biol 
            (Oxf)."; date: "2022-05-3"; volume: "7(1):ysac007. doi"; pages: " 
            10.1093/synbio/ysac007. eCollection 2022"
COMMENT     Sequence Label: pCas9CyMutL
FEATURES             Location/Qualifiers
     source          1..9659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(225..242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(225..241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(281..308)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(324..395)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(422..454)
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AANDENYALAA"
     CDS             complement(455..4558)
                     /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"
     CDS             complement(455..4558)
                     /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"
     promoter        4594..4649
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4595..4613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        4650..4684
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     CDS             4697..5323
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(4773..4792)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        5353..5455
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5456..6115
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5522..5541)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      6136..6230
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6344..6889
                     /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.
                     "
     primer_bind     6790..6809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_RNA        complement(8943..8992)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    8996..9023
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    9054..9081
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
ORIGIN
        1 gttgaacgta atcgttttgt tgttgaagat atgtggctgg gtgtgctggt gagccgtggt
       61 ctgagccgtg atgatgccga agatattctg tggctgattt ttaacagcgt tcgtggtctg
      121 gtagttcgta gcctgtggca gaaagataaa gaacgttttg aacgtgtgcg taatagcacc
      181 ctggaaattg cacgtgaacg ttatgcaaaa ttcaaacgtt gatcactggc cgtcgtttta
      241 caacgtcgtg acggaagcgg aatatatccc taggtataaa cgcagaaagg cccacccgaa
      301 ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag
      361 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggagatcct tactcgagtt
      421 aagcagccag agcgtagttt tcgtcgttag cagcgtcacc tcctagctga ctcaaatcaa
      481 tgcgtgtttc ataaagacca gtgatggatt gatggataag agtggcatct aaaacttctt
      541 ttgtagacgt atatcgttta cgatcaattg ttgtatcaaa atatttaaaa gcagcgggag
      601 ctccaagatt cgtcaacgta aataaatgaa taatattttc tgcttgttca cgtattggtt
      661 tgtctctatg tttgttatat gcactaagaa ctttatctaa attggcatct gctaaaataa
      721 cacgcttaga aaattcactg atttgctcaa taatctcatc taaataatgc ttatgctgct
      781 ccacaaacaa ttgtttttgt tcgttatctt ctggactacc cttcaacttt tcataatgac
      841 tagctaaata taaaaaattc acatatttgc ttggcagagc cagctcattt cctttttgta
      901 attctccggc actagccagc atccgtttac gaccgttttc taactcaaaa agactatatt
      961 taggtagttt aatgattaag tcttttttaa cttccttata tcctttagct tctaaaaagt
     1021 caatcggatt tttttcaaag gaacttcttt ccataattgt gatccctagt aactctttaa
     1081 cggattttaa cttcttcgat ttcccttttt ccaccttagc aaccactagg actgaataag
     1141 ctaccgttgg actatcaaaa ccaccatatt tttttggatc ccagtctttt ttacgagcaa
     1201 taagcttgtc cgaatttctt tttggtaaaa ttgactcctt ggagaatccg cctgtctgta
     1261 cttctgtttt cttgacaata ttgacttggg gcatggacaa tactttgcgc actgtggcaa
     1321 aatctcgccc tttatcccag acaatttctc cagtttcccc attagtttcg attagagggc
     1381 gtttgcgaat ctctccattt gcaagtgtaa tttctgtttt gaagaagttc atgatattag
     1441 agtaaaagaa atattttgcg gttgctttgc ctatttcttg ctcagactta gcaatcattt
     1501 tacgaacatc ataaacttta taatcaccat agacaaactc cgattcaagt tttggatatt
     1561 tcttaatcaa agcagttcca acgacggcat ttagatacgc atcatgggca tgatggtaat
     1621 tgttaatctc acgtacttta tagaattgga aatcttttcg gaagtcagaa actaatttag
     1681 attttaaggt aatcacttta acctctcgaa taagtttatc attttcatcg tatttagtat
     1741 tcatgcgact atccaaaatt tgtgccacat gcttagtgat ttggcgagtt tcaaccaatt
     1801 ggcgtttgat aaaaccagct ttatcaagtt cactcaaacc tccacgttca gctttcgtta
     1861 aattatcaaa cttacgttga gtgattaact tggcgtttag aagttgtctc caatagtttt
     1921 tcatcttttt gactacttct tcacttggaa cgttatccga tttaccacga tttttatcag
     1981 aacgcgttaa gaccttattg tctattgaat cgtctttaag gaaactttgt ggaacaatgt
     2041 gatcgacatc ataatcactt aaacgattaa tatctaattc ttggtccaca tacatgtctc
     2101 ttccattttg gagataatag agatagagct tttcattttg caattgagta ttttcaacag
     2161 gatgctcttt aagaatctga cttcctaatt ctttgatacc ttcttcgatt cgtttcatac
     2221 gctctcgcga atttttctgg cccttttgag ttgtctgatt ttcacgtgcc atttcaataa
     2281 cgatattttc tggcttatgc cgccccatta ctttgaccaa ttcatcaaca acttttacag
     2341 tctgtaaaat acctttttta atagcagggc taccagctaa atttgcaata tgttcatgta
     2401 aactatcgcc ttgtccagac acttgtgctt tttgaatgtc ttctttaaat gtcaaactat
     2461 catcatggat cagctgcata aaattgcgat tggcaaaacc atctgatttc aaaaaatcta
     2521 atattgtttt gccagattgc ttatccctaa taccattaat caattttcga gacaaacgtc
     2581 cccaaccagt ataacggcga cgtttaagct gtttcatcac cttatcatca aagaggtgag
     2641 catatgtttt aagtctttcc tcaatcatct ccctatcttc aaataaggtc aatgttaaaa
     2701 caatatcctc taagatatct tcattttctt cattatccaa aaaatcttta tctttaataa
     2761 tttttagcaa atcatggtag gtacctaatg aagcattaaa tctatcttca actcctgaaa
     2821 tttcaacact atcaaaacat tctatttttt tgaaataatc ttcttttaat tgcttaacgg
     2881 ttacttttcg atttgttttg aagagtaaat caacaatggc tttcttctgt tcacctgaaa
     2941 gaaatgctgg ttttcgcatt ccttcagtaa catatttgac ctttgtcaat tcgttataaa
     3001 ccgtaaaata ctcataaagc aaactatgtt ttggtagtac tttttcattt ggaagatttt
     3061 tatcaaagtt tgtcatgcgt tcaataaatg attgagctga agcaccttta tcgacaactt
     3121 cttcaaaatt ccatggggta attgtttctt cagacttccg agtcatccat gcaaaacgac
     3181 tattgccacg cgccaatgga ccaacataat aaggaattcg aaaagtcaag attttttcaa
     3241 tcttctcacg attgtctttt aaaaatggat aaaagtcttc ttgtcttctc aaaatagcat
     3301 gcagctcacc caagtgaatt tgatggggaa tagagccgtt gtcaaaggtc cgttgcttgc
     3361 gcagcaaatc ttcacgattt agtttcacca ataattcctc agtaccatcc attttttcta
     3421 aaattggttt gataaattta taaaattctt cttggctagc tcccccatca atataacctg
     3481 catatccgtt ttttgattga tcaaaaaaga tttctttata cttttctgga agttgttgtc
     3541 gaactaaagc ttttaaaaga gtcaagtctt gatgatgttc atcgtagcgt ttaatcattg
     3601 aagctgatag gggagcctta gttatttcag tatttactct taggatatct gaaagtaaaa
     3661 tagcatctga taaattctta gctgccaaaa acaaatcagc atattgatct ccaatttgcg
     3721 ccaataaatt atctaaatca tcatcgtaag tatcttttga aagctgtaat ttagcatctt
     3781 ctgccaaatc aaaatttgat ttaaaattag gggtcaaacc caatgacaaa gcaatgagat
     3841 tcccaaataa gccatttttc ttctcaccgg ggagctgagc aatgagattt tctaatcgtc
     3901 ttgatttact caatcgtgca gaaagaatcg ctttagcatc tactccactt gcgttaatag
     3961 ggttttcttc aaataattga ttgtaggttt gtaccaactg gataaatagt ttgtccacat
     4021 cactattatc aggatttaaa tctccctcaa tcaaaaaatg accacgaaac ttaatcatat
     4081 gcgctaaggc caaatagatt aagcgcaaat ccgctttatc agtagaatct accaattttt
     4141 ttcgcagatg atagatagtt ggatatttct catgataagc aacttcatct actatatttc
     4201 caaaaatagg atgacgttca tgcttcttgt cttcttccac caaaaaagac tcttcaagtc
     4261 gatgaaagaa actatcatct actttcgcca tctcatttga aaaaatctcc tgtagataac
     4321 aaatacgatt cttccgacgt gtataccttc tacgagctgt ccgtttgaga cgagtcgctt
     4381 ccgctgtctc tccactgtca aataaaagag cccctataag attttttttg atactgtggc
     4441 ggtctgtatt tcccagaacc ttgaactttt tagacggaac cttatattca tcagtgatca
     4501 ccgcccatcc gacgctattt gtgccgatat ctaagcctat tgagtatttc ttatccatag
     4561 atccgaagtc ctctttagat cttttgaatt cttttctcta tcactgatag ggagtggtaa
     4621 aataactcta tcaacgatag agtgtcaact ttatggctag ctcagtccta ggtacaatgc
     4681 tagcaattag gaattaatga tgtctagatt agataaaagt aaagtgatta acagcgcatt
     4741 agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct
     4801 aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc
     4861 cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag
     4921 ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg
     4981 cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga
     5041 aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact
     5101 cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc
     5161 taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac gacaagctat
     5221 cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat
     5281 atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taagacgtcg atatctggcg
     5341 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     5401 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     5461 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     5521 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     5581 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     5641 tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct
     5701 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     5761 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     5821 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     5881 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     5941 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     6001 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     6061 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga
     6121 tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     6181 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta
     6241 gtaacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     6301 ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
     6361 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
     6421 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
     6481 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
     6541 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
     6601 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
     6661 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
     6721 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
     6781 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     6841 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     6901 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     6961 cacgacaggt ttcccgactg gaaagcgatg cctggagatc cttaccttac tgatatcact
     7021 catctttcag ggcttttatc gccggatgta aatcaacaga ttgtaacaga ccacccggcg
     7081 gcgttttcac aagttgcgga cataaccgtt ccacgtccgc cagcagggtt atggcctgtg
     7141 ccattgacca ctgcgcatgt tcgctcatca gatttcgtgc aatccactgc gcaatattgc
     7201 caggttcgaa tacggactgc ttcgccaggt agcctatcag ttcaggaatc aagatttgta
     7261 aattttgttg gcgtaagggt aaaggcactg ccctgatggt cacatgctgt gcatctgact
     7321 ggaaatcaat acccaattcc gccagggcag actgcgcttt ttctaatgcc gatttttctt
     7381 cggcagaaac ttttagccgc aacggaatca gcagcggctg ggcgcaaacg ggcgcttcac
     7441 ccggcgtcaa ttgtgcctga cgcagccaac gttctgccac tggcaaggat aaaagtgaaa
     7501 tgttgccgtc gcgctccagc aacgcacagt cggaatggac gatagtcagt acccgaccaa
     7561 aactctgact gttcgccgca agtgcaggtt cctgcggttc cggcgctttt aatttttgca
     7621 tcggcgcggg cgtttgcaaa agctggcgat acacttcacc ttgctgtttc tggtagcctg
     7681 gctgcgcatt cggccagggg gcagccggac gactgcctga tgctggcgca ggagtgtagc
     7741 gcggagctac cggctcacga gctgccggtt ctgcaaagtg attgcgcccc gccgccacgc
     7801 ggttttccgg aatggaacgc ggtgcaggtt ggggttcatc gtccagcggt agcggcgttt
     7861 ccagttgctg ttgtagcacg ctcagcacgc cctgatagat aaaatcatgc accagacgcg
     7921 actgatggaa acgcacttcg tgtttggcgg ggtgcacgtt gacgtccacc tgatgtgggt
     7981 cgatctccag atacaacaca aatgccggtt gctgatcggc ccccagtttg tcttcgcagg
     8041 cctggcggat cgcgtgattg atcaggcgat cgcgcatcat gcgaccgttc acgtagcaat
     8101 actgaatttc tgccagtgcg ggcgtggtgt gatttggatc ggccacccag ccgcgtagcg
     8161 tgagatcgcc gtgttgccat tcaatcgcca gcgcttgttc aagaaaagcg gtgccgcaaa
     8221 tcgcgcctaa gcgccgttct ttttgcccgc cttccggcac tgcgcggtac tgacgcacaa
     8281 ttttaccgtt atgcgacagg ttgatcgtga cgtcgaaacg cgccagcgca atgcggcgga
     8341 tgatctcatc aatgtggtta aattcggttt tctcggtgcg caggaatttg cgccgcgccg
     8401 gggtgttgta gaacagatcc agcacctcca gcgtcgtccc cacaggatgc gccgccggtt
     8461 ttaccgtcac gttcatatcg cgcccttcgg cataggcctg ccaggcttcc tgctgttctg
     8521 cggtgcgtga agtgagcgtc aggcgggaaa ccgaactgat actcgccagc gcctcaccgc
     8581 gaaagcccag gctgataatg gcttcgagat cgtccagaga ggcgatttta ctggtggcat
     8641 gacgagccag cgccagcgcc agctcatctt ttttgatacc gcagccgtta tcacgaatgc
     8701 ggataagttt cgccccaccg cgttcgatat caatatcgat acgcgtcgca cctgcatcga
     8761 ggctgttttt cactagttct ttgactaccg acgcaggtcg ctcgaccacc tcacctgcgg
     8821 caatctggtt cgccagttgt ggcggtaaga cctgaattgg catcaatcgt ccttagagtt
     8881 ttgcgagtta agcagccaga gctgcgctct tcccagttct taaacaaaat tatttgtaga
     8941 ggctgtttcg tcctcacgga ctcatcagac cggaaagcac atccggtgac agctttacaa
     9001 acagaccaga ttgtctgttt gttgaatcta ttatacagaa aaattttcca taatacaaac
     9061 agaccagatt gtctgtttgt tccggtacat tgacatatgt atcccagcca acgatcgtta
     9121 agcgggagac cagaaacaaa aaaaggcccc ccgttaggga ggccttcaat aattggggcg
     9181 gcgcgccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc caatggaata
     9241 ttcatagtaa ataccaaacg gagattctta tgagcccgaa acgtcgtacc caggcagaac
     9301 gtgcaatgga aacccagggt aaactgattg cagcagcact gggtgttctg cgtgaaaaag
     9361 gttatgcagg ttttcgtatt gcagatgttc cgggtgcagc cggtgttagc cgtggtgcac
     9421 agagccatca ttttccgacc aaactggaac tgctgctggc aacctttgaa tggctgtatg
     9481 agcagattac cgaacgtagc cgtgcacgtc tggcaaaact gaaaccggaa gatgatgtta
     9541 ttcagcagat gctggatgat gcagcagaat tttttctgga tgatgatttt agcatcggcc
     9601 tggatctgat tgttgcagca gatcgtgatc cggcactgcg tgaaggtatt cagcgtacc
//
