LOCUS       sequence-275344-       10577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10577)
  AUTHORS   Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E
  TITLE     A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive 
            Bacterial Immunity.
  JOURNAL   Science. 2012 Aug 17;337(6096):816-21. doi: 10.1126/science.1225829.
            Epub 2012 Jun 28.
  PUBMED    22745249
REFERENCE   2  (bases 1 to 10577)
  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"; doi: 
            "10.1126/science.1225829"; journalName: "Science"; date: 
            "2012-08-17- 17"; volume: "337"; issue: "6096"; pages: "816-21"
COMMENT     Sequence Label: pMJ841
FEATURES             Location/Qualifiers
     source          1..10577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2237..2257)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2288..3385)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(2291..2314)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             complement(3389..3406)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3417..3439
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3454..3478
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(3478..3497)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3479..3497)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3581..3600)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3806..3883
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3884..4966
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3903..3922)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5775..5966
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5960..5982)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6068..6207
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6123..6142
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6222..6239)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6393..6981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6473..6492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7092..7111)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7103..7918
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             7103..7918
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(7176..7195)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(8010..8465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8147..8168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8359..8378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(8502..8549)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     8545..8563
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(8616..8633)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(join(8698..10284,10285..10287,10288..10577,
                     1..2197,2198..2200,2201..2227))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 cttttacagt ctgtaaaata ccttttttaa tagcagggct accagctaaa tttgcaatat
       61 gttcatgtaa actatcgcct tgtccagaca cttgtgcttt ttgaatgtct tctttaaatg
      121 tcaaactatc atcatggatc agctgcataa aattgcgatt ggcaaaacca tctgatttca
      181 aaaaatctaa tattgttttg ccagattgct tatccctaat accattaatc aattttcgag
      241 acaaacgtcc ccaaccagta taacggcgac gtttaagctg tttcatcacc ttatcatcaa
      301 agaggtgagc atatgtttta agtctttcct caatcatctc cctatcttca aataaggtca
      361 atgttaaaac aatatcctct aagatatctt cattttcttc attatccaaa aaatctttat
      421 ctttaataat ttttagcaaa tcatggtagg tacctaatga agcattaaat ctatcttcaa
      481 ctcctgaaat ttcaacacta tcaaaacatt ctattttttt gaaataatct tcttttaatt
      541 gcttaacggt tacttttcga tttgttttga agagtaaatc aacaatggct ttcttctgtt
      601 cacctgaaag aaatgctggt tttcgcattc cttcagtaac atatttgacc tttgtcaatt
      661 cgttataaac cgtaaaatac tcataaagca aactatgttt tggtagtact ttttcatttg
      721 gaagattttt atcaaagttt gtcatgcgtt caataaatga ttgagctgaa gcacctttat
      781 cgacaacttc ttcaaaattc catggggtaa ttgtttcttc agacttccga gtcatccatg
      841 caaaacgact attgccacgc gccaatggac caacataata aggaattcga aaagtcaaga
      901 ttttttcaat cttctcacga ttgtctttta aaaatggata aaagtcttct tgtcttctca
      961 aaatagcatg cagctcaccc aagtgaattt gatggggaat agagccgttg tcaaaggtcc
     1021 gttgcttgcg cagcaaatct tcacgattta gtttcaccaa taattcctca gtaccatcca
     1081 ttttttctaa aattggtttg ataaatttat aaaattcttc ttggctagct cccccatcaa
     1141 tataacctgc atatccgttt tttgattgat caaaaaagat ttctttatac ttttctggaa
     1201 gttgttgtcg aactaaagct tttaaaagag tcaagtcttg atgatgttca tcgtagcgtt
     1261 taatcattga agctgatagg ggagccttag ttatttcagt atttactctt aggatatctg
     1321 aaagtaaaat agcatctgat aaattcttag ctgccaaaaa caaatcagca tattgatctc
     1381 caatttgcgc caataaatta tctaaatcat catcgtaagt atcttttgaa agctgtaatt
     1441 tagcatcttc tgccaaatca aaatttgatt taaaattagg ggtcaaaccc aatgacaaag
     1501 caatgagatt cccaaataag ccatttttct tctcaccggg gagctgagca atgagatttt
     1561 ctaatcgtct tgatttactc aatcgtgcag aaagaatcgc tttagcatct actccacttg
     1621 cgttaatagg gttttcttca aataattgat tgtaggtttg taccaactgg ataaatagtt
     1681 tgtccacatc actattatca ggatttaaat ctccctcaat caaaaaatga ccacgaaact
     1741 taatcatatg cgctaaggcc aaatagatta agcgcaaatc cgctttatca gtagaatcta
     1801 ccaatttttt tcgcagatga tagatagttg gatatttctc atgataagca acttcatcta
     1861 ctatatttcc aaaaatagga tgacgttcat gcttcttgtc ttcttccacc aaaaaagact
     1921 cttcaagtcg atgaaagaaa ctatcatcta ctttcgccat ctcatttgaa aaaatctcct
     1981 gtagataaca aatacgattc ttccgacgtg tataccttct acgagctgtc cgtttgagac
     2041 gagtcgcttc cgctgtctct ccactgtcaa ataaaagagc ccctataaga ttttttttga
     2101 tactgtggcg gtctgtattt cccagaacct tgaacttttt agacggaacc ttatattcat
     2161 cagtgatcac cgcccatccg acgctatttg tgccgatagc taagcctatt gagtatttct
     2221 tatccatcga ggctgctccc tggaaataca ggttttcact agtgttgttg ttgttcgagc
     2281 tcgaattagt ctgcgcgtct ttcagggctt catcgacagt ctgacgaccg ctggcggcgt
     2341 tgatcaccgc agtacgcacg gcataccaga aagcggacat ctgcgggatg ttcggcatga
     2401 tttcaccttt ctgggcgttt tccatagtgg cggcaatacg tggatctttc gccaactctt
     2461 cctcgtaaga cttcagcgct acggcaccca gcggtttgtc tttattaacc gcttccagac
     2521 cttcatcagt cagcagatag ttttcgagga actcttttgc cagctctttg ttcggactgg
     2581 cggcgttaat acctgcgctc agcacgccaa cgaacggttt ggatggttga cccttgaagg
     2641 tcggcagtac cgttacacca taattcactt tgctggtgtc gatgttggac catgcccacg
     2701 ggccgttgat ggtcatcgct gtttcgcctt tattaaaggc agcttctgcg atggagtaat
     2761 cggtgtctgc attcatgtgt ttgtttttaa tcaggtcaac caggaaggtc agacccgctt
     2821 tcgcgccagc gttatccacg cccacgtctt taatgtcgta cttgccgttt tcatacttga
     2881 acgcataacc cccgtcagca gcaatcagcg gccaggtgaa gtacggttct tgcaggttga
     2941 acatcagcgc gctcttacct ttcgctttca gttctttatc cagcgccggg atctcttccc
     3001 aggtttttgg cgggttcggc agcagatctt tgttataaat cagcgataac gcttcaacag
     3061 cgatcgggta agcaatcagc ttgccgttgt aacgtacggc atcccaggta aacggataca
     3121 gcttgtcctg gaacgctttg tccggggtga tttcagccaa caggccagat tgagcgtagc
     3181 caccaaagcg gtcgtgtgcc cagaagataa tgtcagggcc atcgccagtt gccgcaacct
     3241 gtgggaattt ctcttccagt ttatccggat gctcaacggt gactttaatt ccggtatctt
     3301 tctcgaattt cttaccgact tcagcgagac cgttatagcc tttatcgccg ttaatccaga
     3361 ttaccagttt accttcttcg atttttccat ggtgatggtg atggtgcata tgtatatctc
     3421 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
     3481 tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc
     3541 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     3601 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     3661 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
     3721 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     3781 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
     3841 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     3901 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     3961 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     4021 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     4081 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     4141 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     4201 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     4261 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     4321 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     4381 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     4441 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     4501 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     4561 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     4621 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     4681 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     4741 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     4801 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     4861 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     4921 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     4981 atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca
     5041 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     5101 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     5161 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     5221 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     5281 aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga
     5341 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     5401 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     5461 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
     5521 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     5581 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
     5641 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
     5701 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
     5761 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     5821 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     5881 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
     5941 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6001 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     6061 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     6121 tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga
     6181 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     6241 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6301 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6361 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6421 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6481 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6541 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6601 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6661 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6721 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6781 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6841 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6901 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6961 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7021 tggaacgaaa actcacgtta agggattttg gtcatgaaca ataaaactgt ctgcttacat
     7081 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctctaggcc
     7141 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     7201 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     7261 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     7321 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     7381 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     7441 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     7501 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     7561 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     7621 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     7681 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     7741 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     7801 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     7861 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaaga
     7921 attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7981 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     8041 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     8101 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     8161 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     8221 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     8281 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     8341 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     8401 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     8461 cgcgtcccat tcgccaatcc ggatatagtt cctcctttca gcaaaaaacc cctcaagacc
     8521 cgtttagagg ccccaagggg ttatgctagt tattgctcag cggtggcagc agccaactca
     8581 gcttcctttc gggctttgtt agcagccgga tctcagtggt ggtggtggtg gtgctcgagt
     8641 gcggccgcaa gcttgtcgac ggagctcgaa ttcggatcct gcaaagcacc ggcctcgtta
     8701 gtcacctcct agctgactca aatcaatgcg tgtttcataa agaccagtga tggattgatg
     8761 gataagagtg gcatctaaaa cttcttttgt agacgtatat cgtttacgat caattgttgt
     8821 atcaaaatat ttaaaagcag cgggagctcc aagattcgtc aacgtaaata aatgaataat
     8881 attttctgct tgttcacgta ttggtttgtc tctatgtttg ttatatgcac taagaacttt
     8941 atctaaattg gcatctgcta aaataacacg cttagaaaat tcactgattt gctcaataat
     9001 ctcatctaaa taatgcttat gctgctccac aaacaattgt ttttgttcgt tatcttctgg
     9061 actacccttc aacttttcat aatgactagc taaatataaa aaattcacat atttgcttgg
     9121 cagagccagc tcatttcctt tttgtaattc tccggcacta gccagcatcc gtttacgacc
     9181 gttttctaac tcaaaaagac tatatttagg tagtttaatg attaagtctt ttttaacttc
     9241 cttatatcct ttagcttcta aaaagtcaat cggatttttt tcaaaggaac ttctttccat
     9301 aattgtgatc cctagtaact ctttaacgga ttttaacttc ttcgatttcc ctttttccac
     9361 cttagcaacc actaggactg aataagctac cgttggacta tcaaaaccac catatttttt
     9421 tggatcccag tcttttttac gagcaataag cttgtccgaa tttctttttg gtaaaattga
     9481 ctccttggag aatccgcctg tctgtacttc tgttttcttg acaatattga cttggggcat
     9541 ggacaatact ttgcgcactg tggcaaaatc tcgcccttta tcccagacaa tttctccagt
     9601 ttccccatta gtttcgatta gagggcgttt gcgaatctct ccatttgcaa gtgtaatttc
     9661 tgttttgaag aagttcatga tattagagta aaagaaatat tttgcggttg ctttgcctat
     9721 ttcttgctca gacttagcaa tcattttacg aacatcataa actttataat caccatagac
     9781 aaactccgat tcaagttttg gatatttctt aatcaaagca gttccaacga cggcatttag
     9841 atacgcatca tgggcatgat ggtaattgtt aatctcacgt actttataga attggaaatc
     9901 ttttcggaag tcagaaacta atttagattt taaggtaatc actttaacct ctcgaataag
     9961 tttatcattt tcatcgtatt tagtattcat gcgactatcc aaaatttgtg ccacatgctt
    10021 agtgatttgg cgagtttcaa ccaattggcg tttgataaaa ccagctttat caagttcact
    10081 caaacctcca cgttcagctt tcgttaaatt atcaaactta cgttgagtga ttaacttggc
    10141 gtttagaagt tgtctccaat agtttttcat ctttttgact acttcttcac ttggaacgtt
    10201 atccgattta ccacgatttt tatcagaacg cgttaagacc ttattgtcta ttgaatcgtc
    10261 tttaaggaaa ctttgtggaa caatggcatc gacatcataa tcacttaaac gattaatatc
    10321 taattcttgg tccacataca tgtctcttcc attttggaga taatagagat agagcttttc
    10381 attttgcaat tgagtatttt caacaggatg ctctttaaga atctgacttc ctaattcttt
    10441 gataccttct tcgattcgtt tcatacgctc tcgcgaattt ttctggccct tttgagttgt
    10501 ctgattttca cgtgccattt caataacgat attttctggc ttatgccgcc ccattacttt
    10561 gaccaattca tcaacaa
//
