LOCUS       sequence-199733-       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: pMJ825
FEATURES             Location/Qualifiers
     source          1..10577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(918..2000)
                     /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     1962..1981
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2001..2078)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2284..2303
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2387..2406
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2387..2405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2406..2430
                     /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)."
     RBS             2445..2467
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2478..2495
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2499..3596
                     /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     3570..3593
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3627..3647
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3657..3683,3684..3686,3687..7763)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7828..7845
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7898..7916)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7912..7959
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      7996..8451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8083..8102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8293..8314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8543..9358)
                     /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             complement(8543..9358)
                     /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     9266..9285
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9350..9369
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      9480..10068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9969..9988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10222..10239
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10254..10393
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10319..10338)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10479..10501
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(10495..10577,1..109))
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
       61 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
      121 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
      181 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
      241 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
      301 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
      361 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
      421 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
      481 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
      541 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
      601 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
      661 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
      721 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
      781 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
      841 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
      901 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      961 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     1021 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     1081 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     1141 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     1201 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     1261 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     1321 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     1381 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     1441 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     1501 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     1561 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     1621 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     1681 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     1741 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     1801 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     1861 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     1921 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     1981 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     2041 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     2101 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     2161 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     2221 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     2281 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     2341 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     2401 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
     2461 aaggagatat acatatgcac catcaccatc accatggaaa aatcgaagaa ggtaaactgg
     2521 taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg
     2581 agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc
     2641 cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg
     2701 gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca
     2761 agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga
     2821 tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa
     2881 cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga
     2941 tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg
     3001 cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg
     3061 cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca
     3121 ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg
     3181 gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc
     3241 cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg
     3301 ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg
     3361 aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg
     3421 aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc cagaaaggtg
     3481 aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga
     3541 tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactaatt
     3601 cgagctcgaa caacaacaac actagtgaaa acctgtattt ccagggagca gcctcgatgg
     3661 ataagaaata ctcaataggc ttagctatcg gcacaaatag cgtcggatgg gcggtgatca
     3721 ctgatgaata taaggttccg tctaaaaagt tcaaggttct gggaaataca gaccgccaca
     3781 gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca gcggaagcga
     3841 ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt atttgttatc
     3901 tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt catcgacttg
     3961 aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt tttggaaata
     4021 tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg cgaaaaaaat
     4081 tggtagattc tactgataaa gcggatttgc gcttaatcta tttggcctta gcgcatatga
     4141 ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat agtgatgtgg
     4201 acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa aaccctatta
     4261 acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa tcaagacgat
     4321 tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt gggaatctca
     4381 ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg
     4441 ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta ttggcgcaaa
     4501 ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat gctattttac
     4561 tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca gcttcaatga
     4621 ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta gttcgacaac
     4681 aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt
     4741 atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca attttagaaa
     4801 aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg ctgcgcaagc
     4861 aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta
     4921 ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag aagattgaaa
     4981 aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt
     5041 ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg
     5101 tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc
     5161 ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt acggtttata
     5221 acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca tttctttcag
     5281 gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta
     5341 agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag
     5401 gagttgaaga tagatttaat gcttcattag gtacctacca tgatttgcta aaaattatta
     5461 aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat attgttttaa
     5521 cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca tatgctcacc
     5581 tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt tggggacgtt
     5641 tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca atattagatt
     5701 ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat gatgatagtt
     5761 tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat agtttacatg
     5821 aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta cagactgtaa
     5881 aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat atcgttattg
     5941 aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga gagcgtatga
     6001 aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag catcctgttg
     6061 aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagaca
     6121 tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc gatcacattg
     6181 ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg cgttctgata
     6241 aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact
     6301 attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat aatttaacga
     6361 aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg
     6421 ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta
     6481 aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta aaatctaaat
     6541 tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt aacaattacc
     6601 atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt aagaaatatc
     6661 caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt cgtaaaatga
     6721 ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt tactctaata
     6781 tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc
     6841 taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga gattttgcca
     6901 cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca gaagtacaga
     6961 caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag cttattgctc
     7021 gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg gtagcttatt
     7081 cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag
     7141 agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg attgactttt
     7201 tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta cctaaatata
     7261 gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa
     7321 aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta gctagtcatt
     7381 atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc
     7441 ataagcatta tttagatgag attattgagc aaatcagtga attttctaag cgtgttattt
     7501 tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga gacaaaccaa
     7561 tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt ggagctcccg
     7621 ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct acaaaagaag
     7681 ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt
     7741 tgagtcagct aggaggtgac taacgaggcc ggtgctttgc aggatccgaa ttcgagctcc
     7801 gtcgacaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct
     7861 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     7921 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     7981 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     8041 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     8101 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     8161 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     8221 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     8281 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     8341 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     8401 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     8461 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     8521 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     8581 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     8641 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     8701 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     8761 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     8821 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     8881 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     8941 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     9001 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     9061 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     9121 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     9181 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     9241 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     9301 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     9361 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     9421 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     9481 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     9541 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     9601 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     9661 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     9721 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     9781 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     9841 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     9901 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     9961 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
    10021 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
    10081 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
    10141 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
    10201 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
    10261 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
    10321 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
    10381 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
    10441 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
    10501 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
    10561 gtgaagcgat tcacaga
//
