LOCUS       sequence-313187-       12986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expressed dCas9, LacI and TetR, pNBU2 backbone, AmpR,
            TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12986)
  AUTHORS   Taketani M, Zhang J, Zhang S, Triassi AJ, Huang YJ, Griffith LG, 
            Voigt CA
  TITLE     Genetic circuit design automation for the gut resident species 
            Bacteroides thetaiotaomicron.
  JOURNAL   Nat Biotechnol. 2020 Aug;38(8):962-969. doi: 
            10.1038/s41587-020-0468-5. Epub 2020 Mar 30.
  PUBMED    32231334
REFERENCE   2  (bases 1 to 12986)
  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: 
            "10.1038/s41587-020-0468-5"; journalName: "Nat Biotechnol"; date: 
            "2020-08"; volume: "38"; issue: "8"; pages: "962-969"
COMMENT     Sequence Label: pMT423
FEATURES             Location/Qualifiers
     source          1..12986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(30..56,57..59,60..2546,2547..2549,2550..4136)
                     /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
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGEFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4283..5365
                     /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(4302..4321)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5548..6171
                     /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     complement(5621..5640)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(10856..10875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10975..10997
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11035..11053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11117..11221
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11222..11290,11291..12082)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(11440..11459)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            12246..12355
                     /note="incP origin of transfer"
     rep_origin      12469..12857
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 aaataaagac atataaaaga aaagacacca tggataagaa atactcaata ggcttagcta
       61 tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt ccgtctaaaa
      121 agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc
      181 ttttatttga cagtggagag acagcggaag cgactcgtct caaacggaca gctcgtagaa
      241 ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca aatgagatgg
      301 cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg gaagaagaca
      361 agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct tatcatgaga
      421 aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat aaagcggatt
      481 tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat tttttgattg
      541 agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag ttggtacaaa
      601 cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat gctaaagcga
      661 ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct cagctccccg
      721 gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt ttgaccccta
      781 attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca aaagatactt
      841 acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct gatttgtttt
      901 tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga gtaaatactg
      961 aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa catcatcaag
     1021 acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat aaagaaatct
     1081 tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct agccaagaag
     1141 aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag gaattattgg
     1201 tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc
     1261 cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc
     1321 catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcga attccttatt
     1381 atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg aagtctgaag
     1441 aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat
     1501 ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac
     1561 atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc aaatatgtta
     1621 ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatt
     1681 tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa
     1741 aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcat
     1801 taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg gataatgaag
     1861 aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa gatagggaga
     1921 tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg atgaaacagc
     1981 ttaaacgtcg ccgttatact ggttggggac gtttgtctcg aaaattgatt aatggtatta
     2041 gggataagca atctggcaaa acaatattag attttttgaa atcagatggt tttgccaatc
     2101 gcaattttat gcagctgatc catgatgata gtttgacatt taaagaagac attcaaaaag
     2161 cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta gctggtagcc
     2221 ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg gtcaaagtaa
     2281 tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat cagacaactc
     2341 aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat
     2401 taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc
     2461 tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa ttagatatta
     2521 atcgtttaag tgattatgat gtcgatgcca ttgttccaca aagtttcctt aaagacgatt
     2581 caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg gataacgttc
     2641 caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta aacgccaagt
     2701 taatcactca acgtaagttt gataatttaa cgaaagctga acgtggtggt ttgagtgaac
     2761 ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg
     2821 tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttattc
     2881 gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga aaagatttcc
     2941 aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg tatctaaatg
     3001 ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag tttgtctatg
     3061 gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca
     3121 aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta
     3181 cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg gaaactggag
     3241 aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg tccatgcccc
     3301 aagtcaatat tgtcaagaaa acagaagtac agacaggcga attctccaag gagtcaattt
     3361 taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat
     3421 atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct aaggtggaaa
     3481 aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca attatggaaa
     3541 gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat aaggaagtta
     3601 aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa aacggtcgta
     3661 aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct ctgccaagca
     3721 aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt agtccagaag
     3781 ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat gagattattg
     3841 agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta gataaagttc
     3901 ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa aatattattc
     3961 atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt gatacaacaa
     4021 ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt atccatcaat
     4081 ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt gactgatgct
     4141 gtcataggcc ggttgtccgg tagtccggat gatttaagga agatttctct tcttttttca
     4201 ctttctccga acaacctttc ttttaaaatt gttatttttg tagtgatttt aataacctaa
     4261 aatgaatacg atcgtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     4321 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     4381 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     4441 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     4501 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     4561 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     4621 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     4681 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     4741 gacacccatc aacagtatta ttttctccca tgaggacggt acgcgactgg gcgtggagca
     4801 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     4861 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     4921 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     4981 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     5041 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     5101 cgacgatacc gaggacagct catgttatat cccgccgtta accaccatca aacaggattt
     5161 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     5221 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     5281 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5341 ttcccgactg gaaagcgggc agtgaaaaga aaaggcgttt tgtttttctt ctttaccttc
     5401 tttccctttc gctaagagag tctgagaaac gatagaaaaa gaaaagcgaa aaaacttccg
     5461 aaaacatttg gtagttaaaa taaaacctct tacctttgca cccgcgcatt ttaaaataaa
     5521 ataaattatt tatgatatta aacgaatatg tctagattag ataaaagtaa agtgattaac
     5581 agcgcattag agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc
     5641 cagaagctag gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg
     5701 ctcgacgcct tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa
     5761 ggggaaagct ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta
     5821 agtcatcgcg atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa
     5881 actctcgaaa atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta
     5941 tatgcactca gcgctgtggg gcattttact ttaggttgcg tattggaaga tcaagagcat
     6001 caagtcgcta aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga
     6061 caagctatcg aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa
     6121 ttgatcatat gcggattaga aaaacaactt aaatgtgaaa gtgggtctta atgaactgca
     6181 cttgctttga taattaatga taaacaatct aaaagcactc taatcgttat cggagtgctt
     6241 ttagattact aatcaaattg cttctacggc cgcgatcccc cgggctgcag gaattcgata
     6301 tcaagcttat cgataccgtc gactaattgc ctatcttcca gtgatggaac agcatttgtg
     6361 cattggctgc aacaatcagc cttacttgtg cctgttctat ttccgaaccg accgcttgta
     6421 tgaatgcatc aaaattcgtt ttctctacgt tggattcctt gttgctcata ttgtgatgat
     6481 aatttctaca aatatagtca ttggtaacta tctatgaaac tgtttgatac ttttatagtt
     6541 gattaaactt gttcatggca tttgccttaa tatcatccgc tatgtcaatg tagggtttca
     6601 tagctttgta gtcgctgtgt cccgtccatt tcatgaccac ctgtgccggg attccgagag
     6661 ccagcgcatt gcagatgaat gtcctttttc ctgcatgggt actgagcaaa gcgtatttgg
     6721 gtgtgacttc atcaatacgt tcatttccct tgtagtaggt ttcccgtaca ggctcgttga
     6781 tttctgccag ttcgcccagc tctttcaggt aatcgttcat cttctggttg ctgatgacgg
     6841 gcagagccat gtaattctcg aaatggatgt ccttgtattt gtccagtatg gctttgctgt
     6901 atttgttcag ttcaatcgtc aggctgtcgg cagtcttgac tgtggttatt tcgatgtggt
     6961 cggacttcac atcgcttctt ttcagattgc gaacatctga ataccgcaaa ctcgtaaagc
     7021 agcagaacag gaaaacatca cgcacacgtt ccaggtattg cttatccttg ggtatctggt
     7081 agtctttcag cttgttcagt tcatcccaag tcaggaagat tacttttttc gaggtggttt
     7141 tcagtttcgg tttgaacgta tcgtatgcaa tgttctgatg atgtcctttc ttgaagctcc
     7201 agcgcaggaa ccatttgagg aatcccattt gcttgccgat ggtgctgttt ctcatatcct
     7261 tggtgtcacg caggaagttg acgtattcgt tcaatccaaa ctcgttgaaa tagttgaacg
     7321 ttgcatcctc cttgaactct ttgaggtggt tcctcactgc tgcaaatttt tcataggtgg
     7381 atgccgtcca gttattctgg ttaccgcact cttttacaaa ctcatcgaac acctcccaaa
     7441 agctgacagg ggcttcttcc ggctgttctt cactggtgtc tttcattctc atgttgaaag
     7501 cttccttcaa ctgttgggtc gttggcatga cctcctgcac ctcaaattcc ttgaaaatat
     7561 tctggatttc ggcatagtat ttcagcaagt ccgtattgat ttcggctgca ctttgcttta
     7621 gcttgttggt acatccgttc tttacccgct gcttatctgc atcccatttg gctacgtcaa
     7681 tccggtagcc cgttgtaaac tcgatacgtt ggctggcaaa gatgacacgc atacggatgg
     7741 gtacgttctc tacgattggc acaccgttct ttttccggct ctccaatgca aaaatgatgt
     7801 tgcgcttgat attcataatt gggtgcgttt gaaattctac acccaaatat acacccaatt
     7861 attgagatag caaaagacat ttagaaacat ttacttttac tctatattgt aatttacact
     7921 tgattatcag tcgtttgcag ttttatgata ttctgtgaaa gtataagttc gagagcctgt
     7981 ctctccgcaa aaaacgctga aaatcagcag attgcaaaac aaacaccctg ttttacaccc
     8041 aagaatgtaa agtcgggtgt ttttgtttta tttaagataa tacaaccact acataataaa
     8101 agagtagcga tattaaaaga atccgatgag aaaagactaa tatttatcta tccattcagt
     8161 ttgatttctc aggactttac atcgtcctga aagtatttgt tggtaccgag gacgcgttat
     8221 ctccttaacg tacgttttcg ttccattggc cctcaaaccc cgttatatac attcatgtcc
     8281 atttatgtaa aaaatcctgc tgaccttgtt tatgtcttgt cagtcaccat ttgcaaaacc
     8341 atatttgacc ctcaaagagg ctgaatttga taagcaactt gctacatact cataataagg
     8401 agctaaatag aacacgaatg ggaaatactc aaatgccaaa ctaaagaaga tattggccaa
     8461 aataaacgct ataccgagag agaaacttga tttttcaact tcctaaaaca gtgttgttca
     8521 aacatttcta cttatttgta cttaccagtt gaacctacgt ttccctaata aaatgtctat
     8581 ggtaaaaagt taaaaaatcc tcctactttt gttagatata tttttttgtg taattttgta
     8641 atcgttatgc ggcagtaata atatacatat taatacgagt taggaatcct gtagttctca
     8701 tatgctacga ggaggtatta aaaggtgcgt ttcgacaatg catctattgt agtatattat
     8761 tgcttaatcc aaatgaatat tataaattta ggaattcttg ctcacattga tgcaggaaaa
     8821 acttccgtaa ccgagaatct gctgtttgcc agtggagcaa cggaaaagtg cggctgtgtg
     8881 gataatggtg acaccataac ggactctatg gatatagaga aacgtagagg aattactgtt
     8941 cgggcttcta cgacatctat tatctggaat ggtgtgaaat gcaatatcat tgacactccg
     9001 ggacacatgg attttattgc ggaagtggag cggacattca aaatgcttga tggagcagtc
     9061 ctcatcttat ccgcaaagga aggcatacaa gcgcagacaa agttgctgtt caatacttta
     9121 cagaagctgc aaatcccgac aattatattt atcaataaga ttgaccgagc cggtgtgaat
     9181 ttggagcgtt tgtatctgga tataaaagca aatctgtctc aagatgtcct gtttatgcaa
     9241 aatgttgtcg atggatcggt ttatccggtt tgctcccaaa catatataaa ggaagaatac
     9301 aaagaatttg tatgcaacca tgacgacaat atattagaac gatatttggc ggatagcgaa
     9361 atttcaccgg ctgattattg gaatacgata atcgctcttg tggcaaaagc caaagtctat
     9421 ccggtgctac atggatcagc aatgttcaat atcggtatca atgagttgtt ggacgccatc
     9481 acttctttta tacttcctcc ggcatcggtc tcaaacagac tttcatctta tctttataag
     9541 atagagcatg accccaaagg acataaaaga agttttctaa aaataattga cggaagtctg
     9601 agacttcgag acgttgtaag aatcaacgat tcggaaaaat tcatcaagat taaaaatcta
     9661 aaaactatca atcagggcag agagataaat gttgatgaag tgggcgccaa tgatatcgcg
     9721 attgtagagg atatggatga ttttcgaatc ggaaattatt taggtgctga accttgtttg
     9781 attcaaggat tatcgcatca gcatcccgct ctcaaatcct ccgtccggcc agacaggccc
     9841 gaagagagaa gcaaggtgat atccgctctg aatacattgt ggattgaaga cccgtctttg
     9901 tccttttcca taaactcata tagtgatgaa ttggaaatct cgttatatgg tttaacccaa
     9961 aaggaaatca tacagacatt gctggaagaa cgattttccg taaaggtcca ttttgatgag
    10021 atcaagacta tatacaaaga acgacctgta aaaaaggtca ataagattat tcagatcgaa
    10081 gtgccgccca acccttattg ggccacaata gggctgactc ttgaaccctt accgttaggg
    10141 acagggttgc aaatcgaaag tgacatctcc tatggttatc tgaaccattc ttttcaaaat
    10201 gccgtttttg aagggattcg tatgtcttgc caatccgggt tacatggatg ggaagtgact
    10261 gatctgaaag taacttttac tcaagccgag tattatagcc cggtaagtac acctgctgat
    10321 ttcagacagc tgacccctta tgtcttcagg ctggccttgc aacagtcagg tgtggacatt
    10381 ctcgaaccga tgctctattt tgagttgcag ataccccaag cggcaagttc caaagctatt
    10441 acagatttgc aaaaaatgat gtctgagatt gaagacatca gttgcaataa tgagtggtgt
    10501 catattaaag ggaaagttcc attaaataca agtaaagact atgcatcaga agtaagttca
    10561 tacactaagg gcttaggcat ttttatggtt aagccatgcg ggtatcaaat aacaaaaggc
    10621 ggttattctg ataatatccg catgaacgaa aaagataaac ttttattcat gttccaaaaa
    10681 tcaatgtcat caaaataatg gagcggtcag gaaatttcta taaggcaata cagttgggat
    10741 atatacttat ctccattctt atcggatgta tggcatataa tagcctctat gaatggcagg
    10801 agatagaagc attagaactt ggcaataaaa aaatagacgt atggtgcact ctcagtacaa
    10861 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
    10921 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
    10981 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
    11041 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag caggtggcac
    11101 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
    11161 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
    11221 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
    11281 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
    11341 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
    11401 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
    11461 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
    11521 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
    11581 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
    11641 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
    11701 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
    11761 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
    11821 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
    11881 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
    11941 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
    12001 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
    12061 ctcactgatt aagcattggt aaacgcgtca attcgagggg gatcaattcc gttttccgct
    12121 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
    12181 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
    12241 cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc
    12301 ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc
    12361 gccggcgtaa cagatgaggg caagcggatg gctgatgaaa ccaagccaac aggaagggca
    12421 gcccacctat cacggaattg atccggccac gatgcgtccg gcgtagagga tctgaagatc
    12481 agcagttcaa cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca
    12541 tgtttaacgt actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag
    12601 ctctcatggc taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca
    12661 ataaaattaa tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa
    12721 aaaaagaata tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca
    12781 gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga
    12841 acacttaacg gctgacatgg gaattcccct ccaccgcggt ggctcggtac caaagacgaa
    12901 caataagacg ctgaaaagcg tcttttttcg ttttggtccg ataaagtttg gaagataaag
    12961 ctaaaagttc ttatctttgc agtccg
//
