LOCUS       sequence-333575-       13152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9, beta, and mutL mutant expression plasmid for introducing 
            chromosomal point mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13152)
  AUTHORS   Cook TB, Jacobson TB, Venkataraman MV, Hofstetter H, Amador-Noguez 
            D, Thomas MG, Pfleger BF
  TITLE     Stepwise genetic engineering of Pseudomonas putida enables robust 
            heterologous production of prodigiosin and glidobactin A.
  JOURNAL   Metab Eng. 2021 Jun 24;67:112-124. doi: 10.1016/j.ymben.2021.06.004.
  PUBMED    34175462
REFERENCE   2  (bases 1 to 13152)
  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: "Metab 
            Eng."; date: "2021-06-24"; volume: "67:112-124. doi"; pages: " 
            10.1016/j.ymben.2021.06.004"
COMMENT     Sequence Label: pCas9-beta-mmr
FEATURES             Location/Qualifiers
     source          1..13152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      97..143
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    193..214
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        215..249
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     CDS             285..4388
                     /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             285..4388
                     /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             4398..4421
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        join(4636..4641,4642..4658,4659..4664)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4853..5386
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEVVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             6627..7775
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(9799..10584)
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     RBS             10593..10601
                     /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)"
     promoter        complement(10628..10912)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(10686..10705)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             10939..11817
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      complement(11868..11899)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(12033..12656)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     12564..12583
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(12657..12734)
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    12791..12809
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12816..12834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        12865..12940
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             12961..12990
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             13006..13023
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(13079..13096)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
       61 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
      121 aaagactggg cctttcgttt tatctgttgt ttgtcggtga actggatcct tactcgagtc
      181 tagactgcag ttgaattcgc ggccgcttct agagtttacg gctagctcag tcctaggtac
      241 tatgctagct atcagcagga cgcactgacc aggaggtaca atcaatggat aagaaatact
      301 caataggctt agatatcggc acaaatagcg tcggatgggc ggtgatcact gatgaatata
      361 aggttccgtc taaaaagttc aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa
      421 atcttatagg ggctctttta tttgacagtg gagagacagc ggaagcgact cgtctcaaac
      481 ggacagctcg tagaaggtat acacgtcgga agaatcgtat ttgttatcta caggagattt
      541 tttcaaatga gatggcgaaa gtagatgata gtttctttca tcgacttgaa gagtcttttt
      601 tggtggaaga agacaagaag catgaacgtc atcctatttt tggaaatata gtagatgaag
      661 ttgcttatca tgagaaatat ccaactatct atcatctgcg aaaaaaattg gtagattcta
      721 ctgataaagc ggatttgcgc ttaatctatt tggccttagc gcatatgatt aagtttcgtg
      781 gtcatttttt gattgaggga gatttaaatc ctgataatag tgatgtggac aaactattta
      841 tccagttggt acaaacctac aatcaattat ttgaagaaaa ccctattaac gcaagtggag
      901 tagatgctaa agcgattctt tctgcacgat tgagtaaatc aagacgatta gaaaatctca
      961 ttgctcagct ccccggtgag aagaaaaatg gcttatttgg gaatctcatt gctttgtcat
     1021 tgggtttgac ccctaatttt aaatcaaatt ttgatttggc agaagatgct aaattacagc
     1081 tttcaaaaga tacttacgat gatgatttag ataatttatt ggcgcaaatt ggagatcaat
     1141 atgctgattt gtttttggca gctaagaatt tatcagatgc tattttactt tcagatatcc
     1201 taagagtaaa tactgaaata actaaggctc ccctatcagc ttcaatgatt aaacgctacg
     1261 atgaacatca tcaagacttg actcttttaa aagctttagt tcgacaacaa cttccagaaa
     1321 agtataaaga aatctttttt gatcaatcaa aaaacggata tgcaggttat attgatgggg
     1381 gagctagcca agaagaattt tataaattta tcaaaccaat tttagaaaaa atggatggta
     1441 ctgaggaatt attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg
     1501 acaacggctc tattccccat caaattcact tgggtgagct gcatgctatt ttgagaagac
     1561 aagaagactt ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt
     1621 ttcgaattcc ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga
     1681 ctcggaagtc tgaagaaaca attaccccat ggaattttga agaagttgtc gataaaggtg
     1741 cttcagctca atcatttatt gaacgcatga caaactttga taaaaatctt ccaaatgaaa
     1801 aagtactacc aaaacatagt ttgctttatg agtattttac ggtttataac gaattgacaa
     1861 aggtcaaata tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga
     1921 aagccattgt tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag
     1981 aagattattt caaaaaaata gaatgttttg atagtgttga aatttcagga gttgaagata
     2041 gatttaatgc ttcattaggt acctaccatg atttgctaaa aattattaaa gataaagatt
     2101 ttttggataa tgaagaaaat gaagatatct tagaggatat tgttttaaca ttgaccttat
     2161 ttgaagatag ggagatgatt gaggaaagac ttaaaacata tgctcacctc tttgatgata
     2221 aggtgatgaa acagcttaaa cgtcgccgtt atactggttg gggacgtttg tctcgaaaat
     2281 tgattaatgg tattagggat aagcaatctg gcaaaacaat attagatttt ttgaaatcag
     2341 atggttttgc caatcgcaat tttatgcagc tgatccatga tgatagtttg acatttaaag
     2401 aagacattca aaaagcacaa gtgtctggac aaggcgatag tttacatgaa catattgcaa
     2461 atttagctgg tagccctgct attaaaaaag gtattttaca gactgtaaaa gttgttgatg
     2521 aattggtcaa agtaatgggg cggcataagc cagaaaatat cgttattgaa atggcacgtg
     2581 aaaatcagac aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag
     2641 aaggtatcaa agaattagga agtcagattc ttaaagagca tcctgttgaa aatactcaat
     2701 tgcaaaatga aaagctctat ctctattatc tccaaaatgg aagagacatg tatgtggacc
     2761 aagaattaga tattaatcgt ttaagtgatt atgatgtcga tcacattgtt ccacaaagtt
     2821 tccttaaaga cgattcaata gacaataagg tcttaacgcg ttctgataaa aatcgtggta
     2881 aatcggataa cgttccaagt gaagaagtag tcaaaaagat gaaaaactat tggagacaac
     2941 ttctaaacgc caagttaatc actcaacgta agtttgataa tttaacgaaa gctgaacgtg
     3001 gaggtttgag tgaacttgat aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc
     3061 aaatcactaa gcatgtggca caaattttgg atagtcgcat gaatactaaa tacgatgaaa
     3121 atgataaact tattcgagag gttaaagtga ttaccttaaa atctaaatta gtttctgact
     3181 tccgaaaaga tttccaattc tataaagtac gtgagattaa caattaccat catgcccatg
     3241 atgcgtatct aaatgccgtc gttggaactg ctttgattaa gaaatatcca aaacttgaat
     3301 cggagtttgt ctatggtgat tataaagttt atgatgttcg taaaatgatt gctaagtctg
     3361 agcaagaaat aggcaaagca accgcaaaat atttctttta ctctaatatc atgaacttct
     3421 tcaaaacaga aattacactt gcaaatggag agattcgcaa acgccctcta atcgaaacta
     3481 atggggaaac tggagaaatt gtctgggata aagggcgaga ttttgccaca gtgcgcaaag
     3541 tattgtccat gccccaagtc aatattgtca agaaaacaga agtacagaca ggcggattct
     3601 ccaaggagtc aattttacca aaaagaaatt cggacaagct tattgctcgt aaaaaagact
     3661 gggatccaaa aaaatatggt ggttttgata gtccaacggt agcttattca gtcctagtgg
     3721 ttgctaaggt ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag ttactaggga
     3781 tcacaattat ggaaagaagt tcctttgaaa aaaatccgat tgacttttta gaagctaaag
     3841 gatataagga agttaaaaaa gacttaatca ttaaactacc taaatatagt ctttttgagt
     3901 tagaaaacgg tcgtaaacgg atgctggcta gtgccggaga attacaaaaa ggaaatgagc
     3961 tggctctgcc aagcaaatat gtgaattttt tatatttagc tagtcattat gaaaagttga
     4021 agggtagtcc agaagataac gaacaaaaac aattgtttgt ggagcagcat aagcattatt
     4081 tagatgagat tattgagcaa atcagtgaat tttctaagcg tgttatttta gcagatgcca
     4141 atttagataa agttcttagt gcatataaca aacatagaga caaaccaata cgtgaacaag
     4201 cagaaaatat tattcattta tttacgttga cgaatcttgg agctcccgct gcttttaaat
     4261 attttgatac aacaattgat cgtaaacgat atacgtctac aaaagaagtt ttagatgcca
     4321 ctcttatcca tcaatccatc actggtcttt atgaaacacg cattgatttg agtcagctag
     4381 gaggtgacgg cagcggtgat tataaagatg atgatgataa atgagatatc attcaggacg
     4441 agcctcagac tccagcgtaa ctggactgcg catgcataaa aactgttgta attcattaag
     4501 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     4561 cagcaccttg tcgccttgcg tataatattt gcccatggac gcacaccgtg gaaacggatg
     4621 aaggcacgaa cccagttgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt
     4681 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     4741 gcggttttca tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag
     4801 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
     4861 cagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag gtggctcaag
     4921 tatgggcatc attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc
     4981 tcttgatctt ttcggtcgtg agttcggaga cgtagccacc tactcccaac atcagccgga
     5041 ctccgattac ctcgggaact tgctccgtag taagacattc atcgcgcttg ctgccttcga
     5101 ccaagaagtg gttgttggcg ctctcgcggc ttacgttctg cccaagtttg agcagccgcg
     5161 tagtgagatc tatatctatg atctcgcagt ctccggagag caccggaggc agggcattgc
     5221 caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta
     5281 cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg
     5341 ggaagaagtg atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc
     5401 cgagatcggc ttcccggccg cggagttgtt cggtaaattg tcacaacgcc gcaggtggca
     5461 cttttcgggg aaatgtgcgc gcccgcgttc ctgctggcgc tgggcctgtt tctggcgctg
     5521 gacttcccgc tgttccgtca gcagcttttc gcccacggcc ttgatgatcg cggcggcctt
     5581 ggcctgcata tcccgattca acggccccag ggcgtccaga acgggcttca ggcgctcccg
     5641 aaggtggatc cccccctcgc attggtaact gtcagaccaa ggtgagaatc caagcactag
     5701 tgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcggggga tcaggaccgc
     5761 tgccggagcg caacccactc actacagcag agccatgtag ggccgccggc gttgtggata
     5821 cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca cttgaggggc
     5881 cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc cggcgacgtg
     5941 gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat
     6001 gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg gcgcgactac
     6061 tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca gatgaggggc
     6121 gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt gcaagggttt
     6181 ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac caatatttat
     6241 aaaccttgtt tttaaccagg gctgcgcccc tcggccgcga acggcctcac cccaaaaatg
     6301 gcagccacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     6361 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     6421 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     6481 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     6541 ccgccaagga tctgatggcg caggggatca agatcgacgg atcgatccgg ggaattaatt
     6601 ccggggcaat cccgcaagga gggtgaatga atcggacgtt tgaccggaag gcatacaggc
     6661 aagaactgat cgacgcgggg ttttccgccg aggatgccga aaccatcgca agccgcaccg
     6721 tcatgcgtgc gccccgcgaa accttccagt ccgtcggctc gatagtccag caagctacgg
     6781 ccaagatcga gcgcgacagc gtgcaactgg ctccccctgc cctgcccgcg ccatcggccg
     6841 ccgtggagcg ttcgcgtcgt ctcgaacagg aggcggcagg tttggcgaag tcgatgacca
     6901 tcgacacgcg aggaactatg acgaccaaga agcgaaaaac cgccggcgag gacctggcaa
     6961 aacaggtcag cgaagccaag caggccgcgt tgctgaaaca cacgaagcag cagatcaagg
     7021 aaatgcagct ttccttgttc gatattgcgc cgtggccgga cacgatgcga gcgatgccaa
     7081 acgacacggc ccgctctgcc ctgttcacca cgcgcaacaa gaaaatcccg cgcgaggcgc
     7141 tgcaaaacaa ggtcattttc cacgtcaaca aggacgtgaa gatcacctac accggcgtcg
     7201 agctgcgggc cgacgatgac gaactggtgt ggcagcaggt gttggagtac gcgaagcgca
     7261 cccctatcgg cgagccgatc accttcacgt tctacgagct ttgccaggac ctgggctggt
     7321 cgatcaatgg ccggtattac acgaaggccg aggaatgcct gtcgcgccta caggcgacgg
     7381 cgatgggctt cacgtccgac cgcgttgggc acctggaatc ggtgtcgctg ctgcaccgct
     7441 tccgcgtcct ggaccgtggc aagaaaacgt cccgttgcca ggtcctgatc gacgaggaaa
     7501 tcgtcgtgct gtttgctggc gaccactaca cgaaattcat atgggagaag taccgcaagc
     7561 tgtcgccgac ggcccgacgg atgttcgact atttcagctc gcaccgggag ccgtacccgc
     7621 tcaagctgga aaccttccgc ctcatgtgcg gatcggattc cacccgcgtg aagaagtggc
     7681 gcgagcaggt cggcgaagcc tgcgaagagt tgcgaggcag cggcctggtg gaacacgcct
     7741 gggtcaatga tgacctggtg cattgcaaac gctagggcct tgtggggtca gttccggctg
     7801 ggggttcagc agccacctgc atcgccctag gtataaaggg aaaactgtcc atacccatgg
     7861 accgcaagct ttcatcgacc gcgcaggaaa agtttgtcca gatcgtccag ccccatctgg
     7921 gtccaggtag gacgaccatg gttgcactgg ccgctgcgct cggtgttttc catgtcgcgc
     7981 agcagggcgt tcatctcggg aattgccaga cgccggttgg cgcgcacggc accgtggcaa
     8041 gccatggtgc cgagcagctc attgaggtgc gcctgaatgc gatcactggt gccgtactcc
     8101 ataaggtcgg caagcacgtc ctgcaccagg cggttggctt cagcctgctt gagcaaggcc
     8161 gggatctggc gaatcgccag ggtctcgggg cccaggcgct gcaattcgaa gcccaggcgc
     8221 tgaaaccact gtgcatgctc ttcggcacag tcggcttcac gctggctgag ggccagggtc
     8281 tccggcacca gcagcggctg gccgctcagg ccctcgctgg ccatggccac cttgaggcgc
     8341 tcgtacatta tccgctcgtg ggcggcgtgc atgtccacca gcaccaggcc aacagcattc
     8401 tcggccagga tgtagatacc cttgagctgc gccagggcat agcctagcgg gggaatgtcg
     8461 ccctggcttt cgggcagtgc cgtcgcaggc gcagcacctg tttccagcgg cttatagaac
     8521 tcgcgataca ccgcctgtgc ctctgcggcc ggcagcggct gcgagggacg tggggtgtat
     8581 tgatactggt aaccggcgcc gctgccacca ttggaaaacg actgttgcgg ggctcgaggc
     8641 tgttcaagca ccggcgaggc caggcgcatt tcaccctggg gcccgaactc accggcctgc
     8701 tggccggtgg ggcgaaccag ctcaggtacg gcagcaggtg cggccagctg gtcttccgga
     8761 cgtacatcgg ccaaggcacg gtgcaaggtg ccatacagga agtcatgcac cgagcgccct
     8821 tcgcggaacc gtacttcatg cttggttggg tgcacgttca catcgacgcc gttgggctcc
     8881 agttccagga acagcacgaa cgtggggtgg cggccgttga acagcacgtc ccggtaggcc
     8941 tgacgcacgg catgggcgac cagtttgtcg cgcactgccc gaccattgac gaagaagtac
     9001 tgcaggtctg cctggctgcg cgagaacgtc ggcaggccga cccagcccca aaggcgcaga
     9061 ccgttgcgct cgacgtcgat cggcagtgcc tgctccatga agcccgggcc gcagatggca
     9121 cccacccgcc gtgcacgggc gatttcgtcg tgggcctcgt gcaggctgag gatgctcttg
     9181 ccattgtggc gcaggtggaa gccgacatcg aaacgggcca gcgccagccg ccggatcact
     9241 tcctgcaagt gatcgaattc ggttttctcg gctttgagga acttgcgcct ggccggggta
     9301 ttgaagaaca aatcgcgcac ttccaccgag gtaccgaccg ggtgcgccgc cggctgcacc
     9361 cgcggcgtca tgtcgcggcc ttcggtttct acttgccagg cctcactggc actggcggta
     9421 cgcgaggtca gggtcaggcg cgccaccgaa ctgatcgagg ccaaggcctc gccacggaag
     9481 cccaggctca ataccccttc taggtcttcc agctcgcgga tcttgctggt ggcgtgtcga
     9541 gccagggcca gcggcaggtc gtcggcagaa atgccgctgc catcatcccg caccttcagc
     9601 agtttgacgc cgccctgctc gacttcgata tcgatgcgcc gggcaccgga gtccaggctg
     9661 tttttcagca gttccttggc cacagacgcc ggccgctcga caacctcgcc ggcggcaatc
     9721 tggttggcca gccgcgggct gagcagctga attcgcgaac cgccactcat ggtatattct
     9781 ccttcttcta gaggatcctc atgctgccac cttctgctct gcggctttct gtttcaggaa
     9841 tccaagagct tttactgctt cggcctgtgt cagttctgac gatgcacgaa tgtcgcggcg
     9901 aaatatctgg gaacagagcg gcaataagtc gtcatcccat gttttatcca gggcgatcag
     9961 cagagtgtta atctcctgca tggtttcatc gttaaccgga gtgatgtcgc gttccggctg
    10021 acgttctgca gtgtatgcag tattttcgac aatgcgctcg gcttcatcct tgtcatagat
    10081 accagcaaat ccgaaggcca gacgggcaca ctgaatcatg gctttatgac gtaacatccg
    10141 tttgggatgc gactgccacg gccccgtgat ttctctgcct tcgcgagttt tgaatggttc
    10201 gcggcggcat tcatccatcc attcggtaac gcagatcgga tgattacggt ccttgcggta
    10261 aatccggcat gtacaggatt cattgtcctg ctcaaagtcc atgccatcaa actgctggtt
    10321 ttcattgatg atgcgggacc agccatcaac gcccaccacc ggaacgatgc cattctgctt
    10381 atcaggaaag gcgtaaattt ctttcgtcca cggattaagg ccgtactggt tggcaacgat
    10441 cagtaatgcg atgaactgcg catcgctggc atcaccttta aatgccgtct ggcgaagagt
    10501 ggtgatcagt tcctgtgggt cgacagaatc catgccgaca cgttcagcca gcttcccagc
    10561 cagcgttgcg agtgcagtac tcatttttta taacctcctt agagctcgaa ttcccaaaaa
    10621 aacgggtatg gagaaacagt agagagttgc gataaaaagc gtcaggtagg atccgctaat
    10681 cttatggata aaaatgctat ggcatagcaa agtgtgacgc cgtgcaaata atcaatgtgg
    10741 acttttctgc cgtgattata gacacttttg ttacgcgttt ttgtcatggc tttggtcccg
    10801 ctttgttaca gaatgctttt aataagcggg gttaccggtt tggttagcga gaagagccag
    10861 taaaagacgc agtgacggca atgtctgatg caatatggac aattggtttc ttctctgaat
    10921 ggcgggagta tgaaaagtat ggctgaagcg caaaatgatc ccctgctgcc gggatactcg
    10981 tttaatgccc atctggtggc gggtttaacg ccgattgagg ccaacggtta tctcgatttt
    11041 tttatcgacc gaccgctggg aatgaaaggt tatattctca atctcaccat tcgcggtcag
    11101 ggggtggtga aaaatcaggg acgagaattt gtttgccgac cgggtgatat tttgctgttc
    11161 ccgccaggag agattcatca ctacggtcgt catccggagg ctcgcgaatg gtatcaccag
    11221 tgggtttact ttcgtccgcg cgcctactgg catgaatggc ttaactggcc gtcaatattt
    11281 gccaatacgg ggttctttcg cccggatgaa gcgcaccagc cgcatttcag cgacctgttt
    11341 gggcaaatca ttaacgccgg gcaaggggaa gggcgctatt cggagctgct ggcgataaat
    11401 ctgcttgagc aattgttact gcggcgcatg gaagcgatta acgagtcgct ccatccaccg
    11461 atggataatc gggtacgcga ggcttgtcag tacatcagcg atcacctggc agacagcaat
    11521 tttgatatcg ccagcgtcgc acagcatgtt tgcttgtcgc cgtcgcgtct gtcacatctt
    11581 ttccgccagc agttagggat tagcgtctta agctggcgcg aggaccaacg tatcagccag
    11641 gcgaagctgc ttttgagcac cacccggatg cctatcgcca ccgtcggtcg caatgttggt
    11701 tttgacgatc aactctactt ctcgcgggta tttaaaaaat gcaccggggc cagcccgagc
    11761 gagttccgtg ccggttgtga agaaaaagtg aatgatgtag ccgtcaagtt gtcataataa
    11821 atcgatgcag gtggcacttt tcggggaaat gtgccggatc cccctcaagt caaaagcctc
    11881 cggtcggagg tttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagtatt
    11941 gagcgatatc tagagaattc gtcaacgaat tcaagcttaa ggttgaaaaa taaaaacggc
    12001 gctaaaaagc gccgtttttt ttgacggtgg tattaagacc cactttcaca tttaagttgt
    12061 ttttctaatc cgcatatgat caattcaagg ccgaataaga aggctggctc tgcaccttgg
    12121 tgatcaaata attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt
    12181 tccctttctt ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa
    12241 tgccccacag cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa
    12301 aaggctaatt gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt
    12361 acttttgctc catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt
    12421 aaaaaatctt gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac
    12481 atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta
    12541 ggctgctcta cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc
    12601 tcattaagca gctctaatgc gctgttaatc actttacttt tatctaatct agacatattc
    12661 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
    12721 cattcgatgg tgtcctcgag ttggtaacga atcagacaat tgacggcttg acggagtagc
    12781 atagctcgag tccctatcag tgatagagat tgacatccct atcagtgata gagatactga
    12841 gcacgagcgc acgagggagc ttccgtttta gagctagaaa tagcaagtta aaataaggct
    12901 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgaa gcttgggccc
    12961 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat
    13021 cattgagttt aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc
    13081 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
    13141 gtagcgcggt gg
//
