LOCUS       sequence-325059-        9681 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 9681)
  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 9681)
  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..9681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /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     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(646..680)
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     promoter        681..736
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    717..735
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             772..4875
                     /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             772..4875
                     /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             4876..4908
                     /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"
     terminator      4935..5006
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5022..5049
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     5088..5105
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5089..5105
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5908..5935
                     /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    5966..5993
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     misc_RNA        5997..6046
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             6108..6116
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     rep_origin      complement(8122..8667)
                     /direction=LEFT
                     /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     complement(8202..8221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(8781..8875)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(8896..9555)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     9470..9489
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(9556..9658)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taattgctag cattgtacct
      661 aggactgagc tagccataaa gttgacactc tatcgttgat agagttattt taccactccc
      721 tatcagtgat agagaaaaga attcaaaaga tctaaagagg acttcggatc tatggataag
      781 aaatactcaa taggcttaga tatcggcaca aatagcgtcg gatgggcggt gatcactgat
      841 gaatataagg ttccgtctaa aaagttcaag gttctgggaa atacagaccg ccacagtatc
      901 aaaaaaaatc ttataggggc tcttttattt gacagtggag agacagcgga agcgactcgt
      961 ctcaaacgga cagctcgtag aaggtataca cgtcggaaga atcgtatttg ttatctacag
     1021 gagatttttt caaatgagat ggcgaaagta gatgatagtt tctttcatcg acttgaagag
     1081 tcttttttgg tggaagaaga caagaagcat gaacgtcatc ctatttttgg aaatatagta
     1141 gatgaagttg cttatcatga gaaatatcca actatctatc atctgcgaaa aaaattggta
     1201 gattctactg ataaagcgga tttgcgctta atctatttgg ccttagcgca tatgattaag
     1261 tttcgtggtc attttttgat tgagggagat ttaaatcctg ataatagtga tgtggacaaa
     1321 ctatttatcc agttggtaca aacctacaat caattatttg aagaaaaccc tattaacgca
     1381 agtggagtag atgctaaagc gattctttct gcacgattga gtaaatcaag acgattagaa
     1441 aatctcattg ctcagctccc cggtgagaag aaaaatggct tatttgggaa tctcattgct
     1501 ttgtcattgg gtttgacccc taattttaaa tcaaattttg atttggcaga agatgctaaa
     1561 ttacagcttt caaaagatac ttacgatgat gatttagata atttattggc gcaaattgga
     1621 gatcaatatg ctgatttgtt tttggcagct aagaatttat cagatgctat tttactttca
     1681 gatatcctaa gagtaaatac tgaaataact aaggctcccc tatcagcttc aatgattaaa
     1741 cgctacgatg aacatcatca agacttgact cttttaaaag ctttagttcg acaacaactt
     1801 ccagaaaagt ataaagaaat cttttttgat caatcaaaaa acggatatgc aggttatatt
     1861 gatgggggag ctagccaaga agaattttat aaatttatca aaccaatttt agaaaaaatg
     1921 gatggtactg aggaattatt ggtgaaacta aatcgtgaag atttgctgcg caagcaacgg
     1981 acctttgaca acggctctat tccccatcaa attcacttgg gtgagctgca tgctattttg
     2041 agaagacaag aagactttta tccattttta aaagacaatc gtgagaagat tgaaaaaatc
     2101 ttgacttttc gaattcctta ttatgttggt ccattggcgc gtggcaatag tcgttttgca
     2161 tggatgactc ggaagtctga agaaacaatt accccatgga attttgaaga agttgtcgat
     2221 aaaggtgctt cagctcaatc atttattgaa cgcatgacaa actttgataa aaatcttcca
     2281 aatgaaaaag tactaccaaa acatagtttg ctttatgagt attttacggt ttataacgaa
     2341 ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac cagcatttct ttcaggtgaa
     2401 cagaagaaag ccattgttga tttactcttc aaaacaaatc gaaaagtaac cgttaagcaa
     2461 ttaaaagaag attatttcaa aaaaatagaa tgttttgata gtgttgaaat ttcaggagtt
     2521 gaagatagat ttaatgcttc attaggtacc taccatgatt tgctaaaaat tattaaagat
     2581 aaagattttt tggataatga agaaaatgaa gatatcttag aggatattgt tttaacattg
     2641 accttatttg aagataggga gatgattgag gaaagactta aaacatatgc tcacctcttt
     2701 gatgataagg tgatgaaaca gcttaaacgt cgccgttata ctggttgggg acgtttgtct
     2761 cgaaaattga ttaatggtat tagggataag caatctggca aaacaatatt agattttttg
     2821 aaatcagatg gttttgccaa tcgcaatttt atgcagctga tccatgatga tagtttgaca
     2881 tttaaagaag acattcaaaa agcacaagtg tctggacaag gcgatagttt acatgaacat
     2941 attgcaaatt tagctggtag ccctgctatt aaaaaaggta ttttacagac tgtaaaagtt
     3001 gttgatgaat tggtcaaagt aatggggcgg cataagccag aaaatatcgt tattgaaatg
     3061 gcacgtgaaa atcagacaac tcaaaagggc cagaaaaatt cgcgagagcg tatgaaacga
     3121 atcgaagaag gtatcaaaga attaggaagt cagattctta aagagcatcc tgttgaaaat
     3181 actcaattgc aaaatgaaaa gctctatctc tattatctcc aaaatggaag agacatgtat
     3241 gtggaccaag aattagatat taatcgttta agtgattatg atgtcgatca cattgttcca
     3301 caaagtttcc ttaaagacga ttcaatagac aataaggtct taacgcgttc tgataaaaat
     3361 cgtggtaaat cggataacgt tccaagtgaa gaagtagtca aaaagatgaa aaactattgg
     3421 agacaacttc taaacgccaa gttaatcact caacgtaagt ttgataattt aacgaaagct
     3481 gaacgtggag gtttgagtga acttgataaa gctggtttta tcaaacgcca attggttgaa
     3541 actcgccaaa tcactaagca tgtggcacaa attttggata gtcgcatgaa tactaaatac
     3601 gatgaaaatg ataaacttat tcgagaggtt aaagtgatta ccttaaaatc taaattagtt
     3661 tctgacttcc gaaaagattt ccaattctat aaagtacgtg agattaacaa ttaccatcat
     3721 gcccatgatg cgtatctaaa tgccgtcgtt ggaactgctt tgattaagaa atatccaaaa
     3781 cttgaatcgg agtttgtcta tggtgattat aaagtttatg atgttcgtaa aatgattgct
     3841 aagtctgagc aagaaatagg caaagcaacc gcaaaatatt tcttttactc taatatcatg
     3901 aacttcttca aaacagaaat tacacttgca aatggagaga ttcgcaaacg ccctctaatc
     3961 gaaactaatg gggaaactgg agaaattgtc tgggataaag ggcgagattt tgccacagtg
     4021 cgcaaagtat tgtccatgcc ccaagtcaat attgtcaaga aaacagaagt acagacaggc
     4081 ggattctcca aggagtcaat tttaccaaaa agaaattcgg acaagcttat tgctcgtaaa
     4141 aaagactggg atccaaaaaa atatggtggt tttgatagtc caacggtagc ttattcagtc
     4201 ctagtggttg ctaaggtgga aaaagggaaa tcgaagaagt taaaatccgt taaagagtta
     4261 ctagggatca caattatgga aagaagttcc tttgaaaaaa atccgattga ctttttagaa
     4321 gctaaaggat ataaggaagt taaaaaagac ttaatcatta aactacctaa atatagtctt
     4381 tttgagttag aaaacggtcg taaacggatg ctggctagtg ccggagaatt acaaaaagga
     4441 aatgagctgg ctctgccaag caaatatgtg aattttttat atttagctag tcattatgaa
     4501 aagttgaagg gtagtccaga agataacgaa caaaaacaat tgtttgtgga gcagcataag
     4561 cattatttag atgagattat tgagcaaatc agtgaatttt ctaagcgtgt tattttagca
     4621 gatgccaatt tagataaagt tcttagtgca tataacaaac atagagacaa accaatacgt
     4681 gaacaagcag aaaatattat tcatttattt acgttgacga atcttggagc tcccgctgct
     4741 tttaaatatt ttgatacaac aattgatcgt aaacgatata cgtctacaaa agaagtttta
     4801 gatgccactc ttatccatca atccatcact ggtctttatg aaacacgcat tgatttgagt
     4861 cagctaggag gtgacgctgc taacgacgaa aactacgctc tggctgctta actcgagtaa
     4921 ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     4981 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     5041 ctttctgcgt ttatacctag ggatatattc cgcttccgtc acgacgttgt aaaacgacgg
     5101 ccagtgatca acgtttgaat tttgcataac gttcacgtgc aatttccagg gtgctattac
     5161 gcacacgttc aaaacgttct ttatctttct gccacaggct acgaactacc agaccacgaa
     5221 cgctgttaaa aatcagccac agaatatctt cggcatcatc acggctcaga ccacggctca
     5281 ccagcacacc cagccacata tcttcaacaa caaaacgatt acgttcaacg gtacgctgaa
     5341 taccttcacg cagtgccgga tcacgatctg ctgcaacaat cagatccagg ccgatgctaa
     5401 aatcatcatc cagaaaaaat tctgctgcat catccagcat ctgctgaata acatcatctt
     5461 ccggtttcag ttttgccaga cgtgcacggc tacgttcggt aatctgctca tacagccatt
     5521 caaaggttgc cagcagcagt tccagtttgg tcggaaaatg atggctctgt gcaccacggc
     5581 taacaccggc tgcacccgga acatctgcaa tacgaaaacc tgcataacct ttttcacgca
     5641 gaacacccag tgctgctgca atcagtttac cctgggtttc cattgcacgt tctgcctggg
     5701 tacgacgttt cgggctcata agaatctccg tttggtattt actatgaata ttccattggg
     5761 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggcgcgccg ccccaattat
     5821 tgaaggcctc cctaacgggg ggcctttttt tgtttctggt ctcccgctta acgatcgttg
     5881 gctgggatac atatgtcaat gtaccggaac aaacagacaa tctggtctgt ttgtattatg
     5941 gaaaattttt ctgtataata gattcaacaa acagacaatc tggtctgttt gtaaagctgt
     6001 caccggatgt gctttccggt ctgatgagtc cgtgaggacg aaacagcctc tacaaataat
     6061 tttgtttaag aactgggaag agcgcagctc tggctgctta actcgtgtaa ggaggttttc
     6121 aaaactctaa ggacgattga tgccaattca ggtcttaccg ccacaactgg cgaaccagat
     6181 tgccgcaggt gaggtggtcg agcgacctgc gtcggtagtc aaagaactag tgaaaaacag
     6241 cctcgatgca ggtgcgacgc gtatcgatat tgatatcgaa cgcggtgggg cgaaacttat
     6301 ccgcattcgt gataacggct gcggtatcaa aaaagatgag ctggcgctgg cgctggctcg
     6361 tcatgccacc agtaaaatcg cctctctgga cgatctcgaa gccattatca gcctgggctt
     6421 tcgcggtgag gcgctggcga gtatcagttc ggtttcccgc ctgacgctca cttcacgcac
     6481 cgcagaacag caggaagcct ggcaggccta tgccgaaggg cgcgatatga acgtgacggt
     6541 aaaaccggcg gcgcatcctg tggggacgac gctggaggtg ctggatctgt tctacaacac
     6601 cccggcgcgg cgcaaattcc tgcgcaccga gaaaaccgaa tttaaccaca ttgatgagat
     6661 catccgccgc attgcgctgg cgcgtttcga cgtcacgatc aacctgtcgc ataacggtaa
     6721 aattgtgcgt cagtaccgcg cagtgccgga aggcgggcaa aaagaacggc gcttaggcgc
     6781 gatttgcggc accgcttttc ttgaacaagc gctggcgatt gaatggcaac acggcgatct
     6841 cacgctacgc ggctgggtgg ccgatccaaa tcacaccacg cccgcactgg cagaaattca
     6901 gtattgctac gtgaacggtc gcatgatgcg cgatcgcctg atcaatcacg cgatccgcca
     6961 ggcctgcgaa gacaaactgg gggccgatca gcaaccggca tttgtgttgt atctggagat
     7021 cgacccacat caggtggacg tcaacgtgca ccccgccaaa cacgaagtgc gtttccatca
     7081 gtcgcgtctg gtgcatgatt ttatctatca gggcgtgctg agcgtgctac aacagcaact
     7141 ggaaacgccg ctaccgctgg acgatgaacc ccaacctgca ccgcgttcca ttccggaaaa
     7201 ccgcgtggcg gcggggcgca atcactttgc agaaccggca gctcgtgagc cggtagctcc
     7261 gcgctacact cctgcgccag catcaggcag tcgtccggct gccccctggc cgaatgcgca
     7321 gccaggctac cagaaacagc aaggtgaagt gtatcgccag cttttgcaaa cgcccgcgcc
     7381 gatgcaaaaa ttaaaagcgc cggaaccgca ggaacctgca cttgcggcga acagtcagag
     7441 ttttggtcgg gtactgacta tcgtccattc cgactgtgcg ttgctggagc gcgacggcaa
     7501 catttcactt ttatccttgc cagtggcaga acgttggctg cgtcaggcac aattgacgcc
     7561 gggtgaagcg cccgtttgcg cccagccgct gctgattccg ttgcggctaa aagtttctgc
     7621 cgaagaaaaa tcggcattag aaaaagcgca gtctgccctg gcggaattgg gtattgattt
     7681 ccagtcagat gcacagcatg tgaccatcag ggcagtgcct ttacccttac gccaacaaaa
     7741 tttacaaatc ttgattcctg aactgatagg ctacctggcg aagcagtccg tattcgaacc
     7801 tggcaatatt gcgcagtgga ttgcacgaaa tctgatgagc gaacatgcgc agtggtcaat
     7861 ggcacaggcc ataaccctgc tggcggacgt ggaacggtta tgtccgcaac ttgtgaaaac
     7921 gccgccgggt ggtctgttac aatctgttga tttacatccg gcgataaaag ccctgaaaga
     7981 tgagtgatat cagtaagtcg tcttggtaag gatctccagg catcgctttc cagtcgggaa
     8041 acctgtcgtg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     8101 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     8161 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     8221 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     8281 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     8341 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     8401 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     8461 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     8521 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     8581 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     8641 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     8701 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     8761 aagttgttac tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     8821 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     8881 gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     8941 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     9001 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     9061 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     9121 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     9181 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     9241 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     9301 ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag
     9361 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     9421 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     9481 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     9541 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     9601 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
     9661 gtctcatttt cgccagatat c
//
