LOCUS Exported 12545 bp ds-DNA circular SYN 24-AUG-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..12545
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA complement(14..89)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 119..135
/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)."
CDS complement(404..1486)
/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
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(1487..1564)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS complement(1698..2648)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 2696..2918
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS complement(3572..4387)
/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"
misc_RNA complement(4543..4621)
/label=tracrRNA
/note="trans-activating CRISPR RNA for the Streptococcus
pyogenes CRISPR/Cas9 system"
CDS 4917..9023
/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(9153..10031)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRH
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 10058..10342
/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)"
CDS 10386..10802
/codon_start=1
/gene="gam"
/product="inhibitor of the host RecBCD nuclease in the
lambda Red system"
/label=Gam
/note="protects the double-stranded DNA substrate for
recombination"
/translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
CDS 10808..11593
/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"
CDS 11590..12270
/codon_start=1
/gene="exo"
/product="5' to 3' double-stranded DNA exonuclease in the
lambda Red system"
/label=Exo
/note="creates 3' single-stranded DNA tails"
/translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
EQWR"
terminator 12271..12515
/label=-lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
ORIGIN
1 gatctcaaaa aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta
61 ttttaacttg ctatttctag ctctaaaacc tggtaacagg attagcagat gtgtgaaatt
121 gttatccgct cacaattcca cacaacatac gagccggatg attaattgtc aacagctcat
181 ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa aaacattatc cagaacggga
241 gtgcgccttg agcgacacga attatgcagt gatttacgac ctgcacagcc ataccacagc
301 ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt gcagtcgatg ataagctgtc
361 aaaccagatc aattcgcgct aacttacatt aattgcgttg cgctcactgc ccgctttcca
421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
481 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
541 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
601 gcaggcgaaa atcctgtttg atggtggttg acggcgggat ataacatgag ctgtcttcgg
661 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
721 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
781 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
841 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
901 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
961 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
1021 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
1081 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
1141 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
1201 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
1261 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
1321 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
1381 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
1441 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
1501 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg
1561 tgtcaacgta aatgcatgcc gcttcgcctt ccatgggtat ggacagtttt ccctttgata
1621 tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata
1681 caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt
1741 cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca
1801 ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt
1861 gtttttctta gtccgttacg taggtaggaa tctgatgtaa tggttgttgg tattttgtca
1921 ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt
1981 tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata
2041 ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta
2101 aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct
2161 atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc
2221 atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt
2281 aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag
2341 aacttggcat agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt
2401 tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc
2461 ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc
2521 cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt
2581 tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat
2641 tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct
2701 agtcaatgat aattactagt ccttttcctt tgagttgtgg gtatctgtaa attctgctag
2761 acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg
2821 tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa
2881 aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag
2941 tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct
3001 aaaggcttaa gtagcaccct cgcaagctcg gttgcggccg caatcgggca aatcgctgaa
3061 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
3121 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
3181 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
3241 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
3301 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
3361 aatgcaccca gtaaggcagc ggtatcatca acggggtctg acgctcagtg gaacgaaaac
3421 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
3481 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
3541 tacgtttcca caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
3601 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
3661 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
3721 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
3781 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
3841 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
3901 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
3961 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
4021 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
4081 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
4141 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
4201 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
4261 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
4321 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
4381 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
4441 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttccc
4501 tgcagggttt gcagtcagag tagaatagaa gtatcaaaaa aagcaccgac tcggtgccac
4561 tttttcaagt tgataacgga ctagccttat tttaacttgc tatgctgttt tgaatggttc
4621 caacaagatt attttataac ttttataaca aataatcaag gagaaattca aagaaattta
4681 tcagccataa aacaatactt aatactatag aatgataaca aaataaacta ctttttaaaa
4741 gaattttgtg ttataatcta tttattatta agtattgggt aatatttttt gaagagatat
4801 tttgaaaaag aaaaattaaa gcatattaaa ctaatttcgg aggtcattaa aactattatt
4861 gaaatcatca aactcattat ggatttaatt taaacttttt attttaggag gcaaaaatgg
4921 ataagaaata ctcaataggc ttagatatcg gcacaaatag cgtcggatgg gcggtgatca
4981 ctgatgaata taaggttccg tctaaaaagt tcaaggttct gggaaataca gaccgccaca
5041 gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca gcggaagcga
5101 ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt atttgttatc
5161 tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt catcgacttg
5221 aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt tttggaaata
5281 tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg cgaaaaaaat
5341 tggtagattc tactgataaa gcggatttgc gcttaatcta tttggcctta gcgcatatga
5401 ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat agtgatgtgg
5461 acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa aaccctatta
5521 acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa tcaagacgat
5581 tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt gggaatctca
5641 ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg
5701 ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta ttggcgcaaa
5761 ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat gctattttac
5821 tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca gcttcaatga
5881 ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta gttcgacaac
5941 aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt
6001 atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca attttagaaa
6061 aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg ctgcgcaagc
6121 aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta
6181 ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag aagattgaaa
6241 aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt
6301 ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg
6361 tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc
6421 ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt acggtttata
6481 acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca tttctttcag
6541 gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta
6601 agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag
6661 gagttgaaga tagatttaat gcttcattag gtacctacca tgatttgcta aaaattatta
6721 aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat attgttttaa
6781 cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca tatgctcacc
6841 tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt tggggacgtt
6901 tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca atattagatt
6961 ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat gatgatagtt
7021 tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat agtttacatg
7081 aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta cagactgtaa
7141 aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat atcgttattg
7201 aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga gagcgtatga
7261 aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag catcctgttg
7321 aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagaca
7381 tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc gatcacattg
7441 ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg cgttctgata
7501 aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact
7561 attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat aatttaacga
7621 aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg
7681 ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta
7741 aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta aaatctaaat
7801 tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt aacaattacc
7861 atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt aagaaatatc
7921 caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt cgtaaaatga
7981 ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt tactctaata
8041 tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc
8101 taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga gattttgcca
8161 cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca gaagtacaga
8221 caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag cttattgctc
8281 gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg gtagcttatt
8341 cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag
8401 agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg attgactttt
8461 tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta cctaaatata
8521 gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa
8581 aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta gctagtcatt
8641 atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc
8701 ataagcatta tttagatgag attattgagc aaatcagtga attttctaag cgtgttattt
8761 tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga gacaaaccaa
8821 tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt ggagctcccg
8881 ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct acaaaagaag
8941 ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt
9001 tgagtcagct aggaggtgac tgaagtatat tttagatgaa gattatttct taatctagac
9061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
9121 tttccccgaa aagtgccacc tgcatcgatt tattatgaca acttgacggc tacatcattc
9181 actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat
9241 acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc
9301 atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt
9361 aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa
9421 acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac
9481 tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc
9541 atgtgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct
9601 tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct
9661 tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc
9721 cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga
9781 cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca
9841 aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa
9901 cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc
9961 gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc
10021 gcttcagcca tacttttcat actcccgcca ttcagagaag aaaccaattg tccatattgc
10081 atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccaa accggtaacc
10141 ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac
10201 aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg cgtcacactt
10261 tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg ctttttatcg
10321 caactctcta ctgtttctcc atacccgttt ttttgggaat tcgagctcta aggaggttat
10381 aaaaaatgga tattaatact gaaactgaga tcaagcaaaa gcattcacta accccctttc
10441 ctgttttcct aatcagcccg gcatttcgcg ggcgatattt tcacagctat ttcaggagtt
10501 cagccatgaa cgcttattac attcaggatc gtcttgaggc tcagagctgg gcgcgtcact
10561 accagcagct cgcccgtgaa gagaaagagg cagaactggc agacgacatg gaaaaaggcc
10621 tgccccagca cctgtttgaa tcgctatgca tcgatcattt gcaacgccac ggggccagca
10681 aaaaatccat tacccgtgcg tttgatgacg atgttgagtt tcaggagcgc atggcagaac
10741 acatccggta catggttgaa accattgctc accaccaggt tgatattgat tcagaggtat
10801 aaaacgaatg agtactgcac tcgcaacgct ggctgggaag ctggctgaac gtgtcggcat
10861 ggattctgtc gacccacagg aactgatcac cactcttcgc cagacggcat ttaaaggtga
10921 tgccagcgat gcgcagttca tcgcattact gatcgttgcc aaccagtacg gccttaatcc
10981 gtggacgaaa gaaatttacg cctttcctga taagcagaat ggcatcgttc cggtggtggg
11041 cgttgatggc tggtcccgca tcatcaatga aaaccagcag tttgatggca tggactttga
11101 gcaggacaat gaatcctgta catgccggat ttaccgcaag gaccgtaatc atccgatctg
11161 cgttaccgaa tggatggatg aatgccgccg cgaaccattc aaaactcgcg aaggcagaga
11221 aatcacgggg ccgtggcagt cgcatcccaa acggatgtta cgtcataaag ccatgattca
11281 gtgtgcccgt ctggccttcg gatttgctgg tatctatgac aaggatgaag ccgagcgcat
11341 tgtcgaaaat actgcataca ctgcagaacg tcagccggaa cgcgacatca ctccggttaa
11401 cgatgaaacc atgcaggaga ttaacactct gctgatcgcc ctggataaaa catgggatga
11461 cgacttattg ccgctctgtt cccagatatt tcgccgcgac attcgtgcat cgtcagaact
11521 gacacaggcc gaagcagtaa aagctcttgg attcctgaaa cagaaagccg cagagcagaa
11581 ggtggcagca tgacaccgga cattatcctg cagcgtaccg ggatcgatgt gagagctgtc
11641 gaacaggggg atgatgcgtg gcacaaatta cggctcggcg tcatcaccgc ttcagaagtt
11701 cacaacgtga tagcaaaacc ccgctccgga aagaagtggc ctgacatgaa aatgtcctac
11761 ttccacaccc tgcttgctga ggtttgcacc ggtgtggctc cggaagttaa cgctaaagca
11821 ctggcctggg gaaaacagta cgagaacgac gccagaaccc tgtttgaatt cacttccggc
11881 gtgaatgtta ctgaatcccc gatcatctat cgcgacgaaa gtatgcgtac cgcctgctct
11941 cccgatggtt tatgcagtga cggcaacggc cttgaactga aatgcccgtt tacctcccgg
12001 gatttcatga agttccggct cggtggtttc gaggccataa agtcagctta catggcccag
12061 gtgcagtaca gcatgtgggt gacgcgaaaa aatgcctggt actttgccaa ctatgacccg
12121 cgtatgaagc gtgaaggcct gcattatgtc gtgattgagc gggatgaaaa gtacatggcg
12181 agttttgacg agatcgtgcc ggagttcatc gaaaaaatgg acgaggcact ggctgaaatt
12241 ggttttgtat ttggggagca atggcgatga cgcatcctca cgataatatc cgggtaggcg
12301 caatcacttt cgtctactcc gttacaaagc gaggctgggt atttcccggc ctttctgtta
12361 tccgaaatcc actgaaagca cagcggctgg ctgaggagat aaataataaa cgaggggctg
12421 tatgcacaaa gcatcttctg ttgagttaag aacgagtatc gagatggcac atagccttgc
12481 tcaaattgga atcaggtttg tgccaatacc agtagaaaca gacgaagaat ccatgggtat
12541 ggaca
//