LOCUS       sequence-357082-       10926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NT-CRISPR plasmid for integration of multiple gRNAs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10926)
  AUTHORS   Stukenberg D, Hoff J, Faber A, Becker A
  TITLE     NT-CRISPR, combining natural transformation and CRISPR-Cas9 
            counterselection for markerless and scarless genome editing in 
            Vibrio natriegens.
  JOURNAL   Commun Biol. 2022 Mar 25;5(1):265. doi: 10.1038/s42003-022-03150-0.
  PUBMED    35338236
REFERENCE   2  (bases 1 to 10926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2022-03-25"; volume: "5(1):265. doi"; pages: " 
            10.1038/s42003-022-03150-0"
COMMENT     Sequence Label: pST_119_LVL2 cam
FEATURES             Location/Qualifiers
     source          1..10926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    61..79
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             176..799
                     /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(249..268)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1076..1110
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             1605..1616
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1623..2336
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2348..2419
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2435..2462
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2511..3099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2591..2610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3206..3240)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3311..3970)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3885..3904
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(4759..5841)
                     /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="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5803..5822
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5918..5923,5924..5939,5940..5946)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5952..5971
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             5980..5991
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(6652..6679)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6695..6766)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6784..6816)
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AANDENYALAA"
     CDS             complement(6823..10926)
                     /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(6823..10926)
                     /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"
ORIGIN
        1 tgattagtcc tgtgtgactc tagtagtgct cagtatctct atcactgata gggatgtcaa
       61 tctctatcac tgatagggag gtcagtgcgt cctgctgaaa acagccgaat ggcgcaaaac
      121 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatatgtc
      181 cagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat
      241 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta
      301 ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag
      361 gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa
      421 cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg
      481 tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca
      541 acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc attttacttt
      601 aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac
      661 tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc
      721 agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa
      781 atgtgaaagt gggtcctgat aaggatccta attggtaacg aatcagaatt gacggctcga
      841 gggagtagca tagggtttgc agaatccctg cttcgtccat ttgacaggca cattatgcat
      901 cgatgataag ctgtcaaaca tgagcagatc ctctacgccg gacgcatcgt ggccggcatc
      961 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     1021 cgggctcgcc acttcgggct catgagcaaa tattttatct gagctccaat gggagtttac
     1081 ggctagctca gtcctaggta tagtgctagc tactagagtc acacaggaaa gtaatcaatg
     1141 aacattaacg acctcatacg agagattaag aacaaagatt acaccgtcaa actgtcagga
     1201 actgatagta actcaatcac ccagcttatt atcagggtaa acaatgatgg gaatgaatat
     1261 gtgatatctg agagcgaaaa cgagtctatc gtcgagaaat tcatttccgc ttttaagaac
     1321 gggtggaatc aggaatatga ggatgaagaa gaattttaca atgacatgca gacgatcacg
     1381 ttgaaaagtg aactgaacta agcttaacgc caaaaacccc gcttcggcgg ggtttttccg
     1441 ccgctgctta gagacggaaa gtgaaacgtg atttcatgcg tcattttgaa cattttgtaa
     1501 atcttattta ataatgtgtg cggcaattca catttaattt atgaatgttt tcttaacatc
     1561 gcggcaactc aagaaacggc aggttcggat cttagctact agagaaagag gagaaatact
     1621 agatgcgtaa aggcgaagaa cttttcactg gcgtagtacc aattttagtg gagctggacg
     1681 gagatgtaaa tggtcataag ttttcagttc gaggagaagg cgaaggagat gcaactaacg
     1741 gtaagctgac actaaagttt atctgtacca cgggtaaact gccggtccca tggccgacac
     1801 tggttactac acttacttat ggtgtacaat gctttgctcg ttatcctgac catatgaagc
     1861 agcacgattt cttcaaatcg gccatgcctg aaggatacgt gcaagaacgt acaattagct
     1921 ttaaagatga cggcacctat aaaacgcgcg ccgaagttaa attcgagggt gacacattgg
     1981 ttaatcgtat agaacttaag ggcattgact tcaaagaaga tggcaacatc cttggccata
     2041 aactggaata taattttaac tctcacaatg tctacattac ggcggataaa caaaagaatg
     2101 gcattaaagc gaatttcaaa atccgccaca atgtcgaaga cggctcggtt caactggcgg
     2161 accattatca gcagaacacc ccaatcggtg atggcccggt tctgttacct gataatcatt
     2221 acctttctac tcaaagcgtt ttatctaaag atcctaacga gaagcgtgat catatggttc
     2281 tactggaatt tgttaccgca gctggtatca cgcacggcat ggatgagctg tataaatgac
     2341 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2401 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     2461 cgtttatacg tctctagctg aaccgtaaaa aaggccgcgt tgctggcgtt tttccacagg
     2521 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2581 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2641 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2701 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2761 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2821 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2881 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2941 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3001 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3061 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3121 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3181 tcaaaaagga tcttcactgc taccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     3241 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
     3301 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     3361 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     3421 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     3481 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgaaacga
     3541 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     3601 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     3661 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     3721 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg
     3781 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     3841 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     3901 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     3961 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     4021 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaaaacag
     4081 gaccaaaacg aaaaaagacg cttttcagcg tcttattgtt cgtctttggt accgagctgt
     4141 gtctgtggct gagagagcag gtacaagata aacccgtcta tattctcaaa ttctttcact
     4201 atcgcacttg ttttatgtga agtccagata tggactctgg aacccaactc ggagctctgc
     4261 gggtagagcg tcaaccattc gaacgttgcg cggtcttcgg agaaccgtaa gaggtaatat
     4321 actccgatat tcatttcagc ggtagctgag tccgtcagtt gatacggtgc gtacccggag
     4381 caaataggag aggcggttgt agtgggggga atggatagca agctgcgggt agacctccaa
     4441 ttattgaagg cctcccaaat cggggggcct tttttattga taacaaaagg agctcagata
     4501 aaatatttgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     4561 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     4621 tagaggatct gctcatgttt gacagcttat catcgatgca taatgtgcct gtcaaatgga
     4681 cgaagcaggg attctgcaaa ccctatgcta ctccctcgag ccgtcaattg tctgattcgt
     4741 taccaattag gatccttatc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4801 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     4861 ttttcttttc accagtgaga ctggcaacag ctgattgccc ttcaccgcct ggccctgaga
     4921 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     4981 ggttaacggc gggatataac atgagctatc ttcggtatcg tcgtatccca ctaccgagat
     5041 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     5101 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     5161 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     5221 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     5281 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     5341 cgtaccgtcc tcatgggagt aaataatact gttgatgggt gtctggtcag agacatcaag
     5401 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     5461 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     5521 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     5581 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     5641 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     5701 gtaattcagc tccaccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     5761 ggcctggttc accacgcggg aaacggtcat ataagagaca ccggcatact ctgcgacatc
     5821 gtataacgtt actggtttca tattcaccac cctgaattga ctctcttccg ggcgctatca
     5881 tgccataccg cgaaaggttt tgcgccattc ggctgtgttg acaattaatc atcggctcgt
     5941 ataatgtgtg gaattgtgag cgctcacaat ttactagaga aagaggagaa ataatcaatg
     6001 gatatgaatg agcaacagtt tttcgactac gtaactaagt ttggcgccta ccaaaaacgc
     6061 tcgatgtttg gtggtattgg cttgtttcaa cacgacgcta tgtatgtgtt ggtcagtgaa
     6121 gaccgcattt ttgtgcgtgg tggagaagag ctcgaccctg agctcttggc cttagggtgc
     6181 gagaaatatc gccatgttaa aaaacagaca acggcaaccg taaactatta cgacatcact
     6241 gaactgtatg agcaacatca ccctgagcta gactcgatta ttgaacgctc gattcagttt
     6301 tctgtgaatc agcgggaatt tcaacgctca gcagccagtc gtcgtctacg ggatttaccc
     6361 aatatgcaac tgactttgga gcgtatggta aaaaaagcgg gcattgacga tgtggaaact
     6421 tttatgagcc tcggtgcgcc agaagtgttt aataaggtgc gtcaagctta tggaagtgat
     6481 gtcgatgtaa aactactgtg gaagtttgcg ggtgcgattg aaggcattca ctggaaactg
     6541 ctgcaagagc cgcgtaagcg ccaactgtta gaaagttgtc agcagcgtgc ttaaaaaaaa
     6601 aaaccccgcc cctgacaggg cggggttttt tttcgcttac tagcgtataa acgcagaaag
     6661 gcccacccga aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga
     6721 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggttaccc
     6781 ctaagcagcc agagcgtagt tttcgtcgtt agcagctaaa gcgtcacctc ctagctgact
     6841 caaatcaatg cgtgtttcat aaagaccagt gatggattga tggataagag tggcatctaa
     6901 aacttctttt gtagacgtat atcgtttacg atcaattgtt gtatcaaaat atttaaaagc
     6961 agcgggagct ccaagattcg tcaacgtaaa taaatgaata atattttctg cttgttcacg
     7021 tattggtttg tctctatgtt tgttatatgc actaagaact ttatctaaat tggcatctgc
     7081 taaaataaca cgcttagaaa attcactgat ttgctcaata atctcatcta aataatgctt
     7141 atgctgctcc acaaacaatt gtttttgttc gttatcttct ggactaccct tcaacttttc
     7201 ataatgacta gctaaatata aaaaattcac atatttgctt ggcagagcca gctcatttcc
     7261 tttttgtaat tctccggcac tagccagcat ccgtttacga ccgttttcta actcaaaaag
     7321 actatattta ggtagtttaa tgattaagtc ttttttaact tccttatatc ctttagcttc
     7381 taaaaagtca atcggatttt tttcaaagga acttctttcc ataattgtga tccctagtaa
     7441 ctctttaacg gattttaact tcttcgattt ccctttttcc accttagcaa ccactaggac
     7501 tgaataagct accgttggac tatcaaaacc accatatttt tttggatccc agtctttttt
     7561 acgagcaata agcttgtccg aatttctttt tggtaaaatt gactccttgg agaatccgcc
     7621 tgtctgtact tctgttttct tgacaatatt gacttggggc atggacaata ctttgcgcac
     7681 tgtggcaaaa tctcgccctt tatcccagac aatttctcca gtttccccat tagtttcgat
     7741 tagagggcgt ttgcgaatct ctccatttgc aagtgtaatt tctgttttga agaagttcat
     7801 gatattagag taaaagaaat attttgcggt tgctttgcct atttcttgct cagacttagc
     7861 aatcatttta cgaacatcat aaactttata atcaccatag acaaactccg attcaagttt
     7921 tggatatttc ttaatcaaag cagttccaac gacggcattt agatacgcat catgggcatg
     7981 atggtaattg ttaatctcac gtactttata gaattggaaa tcttttcgga agtcagaaac
     8041 taatttagat tttaaggtaa tcactttaac ctctcgaata agtttatcat tttcatcgta
     8101 tttagtattc atgcgactat ccaaaatttg tgccacatgc ttagtgattt ggcgagtttc
     8161 aaccaattgg cgtttgataa aaccagcttt atcaagttca ctcaaacctc cacgttcagc
     8221 tttcgttaaa ttatcaaact tacgttgagt gattaacttg gcgtttagaa gttgtctcca
     8281 atagtttttc atctttttga ctacttcttc acttggaacg ttatccgatt taccacgatt
     8341 tttatcagaa cgcgttaaga ccttattgtc tattgaatcg tctttaagga aactttgtgg
     8401 aacaatgtga tcgacatcat aatcacttaa acgattaata tctaattctt ggtccacata
     8461 catgtctctt ccattttgga gataatagag atagagcttt tcattttgca attgagtatt
     8521 ttcaacagga tgctctttaa gaatctgact tcctaattct ttgatacctt cttcgattcg
     8581 tttcatacgc tctcgcgaat ttttctggcc cttttgagtt gtctgatttt cacgtgccat
     8641 ttcaataacg atattttctg gcttatgccg ccccattact ttgaccaatt catcaacaac
     8701 ttttacagtc tgtaaaatac cttttttaat agcagggcta ccagctaaat ttgcaatatg
     8761 ttcatgtaaa ctatcgcctt gtccagacac ttgtgctttt tgaatgtctt ctttaaatgt
     8821 caaactatca tcatggatca gctgcataaa attgcgattg gcaaaaccat ctgatttcaa
     8881 aaaatctaat attgttttgc cagattgctt atccctaata ccattaatca attttcgaga
     8941 caaacgtccc caaccagtat aacggcgacg tttaagctgt ttcatcacct tatcatcaaa
     9001 gaggtgagca tatgttttaa gtctttcctc aatcatctcc ctatcttcaa ataaggtcaa
     9061 tgttaaaaca atatcctcta agatatcttc attttcttca ttatccaaaa aatctttatc
     9121 tttaataatt tttagcaaat catggtaggt acctaatgaa gcattaaatc tatcttcaac
     9181 tcctgaaatt tcaacactat caaaacattc tatttttttg aaataatctt cttttaattg
     9241 cttaacggtt acttttcgat ttgttttgaa gagtaaatca acaatggctt tcttctgttc
     9301 acctgaaaga aatgctggtt ttcgcattcc ttcagtaaca tatttgacct ttgtcaattc
     9361 gttataaacc gtaaaatact cataaagcaa actatgtttt ggtagtactt tttcatttgg
     9421 aagattttta tcaaagtttg tcatgcgttc aataaatgat tgagctgaag cacctttatc
     9481 gacaacttct tcaaaattcc atggggtaat tgtttcttca gacttccgag tcatccatgc
     9541 aaaacgacta ttgccacgcg ccaatggacc aacataataa ggaattcgaa aagtcaagat
     9601 tttttcaatc ttctcacgat tgtcttttaa aaatggataa aagtcttctt gtcttctcaa
     9661 aatagcatgc agctcaccca agtgaatttg atggggaata gagccgttgt caaaggtccg
     9721 ttgcttgcgc agcaaatctt cacgatttag tttcaccaat aattcctcag taccatccat
     9781 tttttctaaa attggtttga taaatttata aaattcttct tggctagctc ccccatcaat
     9841 ataacctgca tatccgtttt ttgattgatc aaaaaagatt tctttatact tttctggaag
     9901 ttgttgtcga actaaagctt ttaaaagagt caagtcttga tgatgttcat cgtagcgttt
     9961 aatcattgaa gctgataggg gagccttagt tatttcagta tttactctta ggatatctga
    10021 aagtaaaata gcatctgata aattcttagc tgccaaaaac aaatcagcat attgatctcc
    10081 aatttgcgcc aataaattat ctaaatcatc atcgtaagta tcttttgaaa gctgtaattt
    10141 agcatcttct gccaaatcaa aatttgattt aaaattaggg gtcaaaccca atgacaaagc
    10201 aatgagattc ccaaataagc catttttctt ctcaccgggg agctgagcaa tgagattttc
    10261 taatcgtctt gatttactca atcgtgcaga aagaatcgct ttagcatcta ctccacttgc
    10321 gttaataggg ttttcttcaa ataattgatt gtaggtttgt accaactgga taaatagttt
    10381 gtccacatca ctattatcag gatttaaatc tccctcaatc aaaaaatgac cacgaaactt
    10441 aatcatatgc gctaaggcca aatagattaa gcgcaaatcc gctttatcag tagaatctac
    10501 caattttttt cgcagatgat agatagttgg atatttctca tgataagcaa cttcatctac
    10561 tatatttcca aaaataggat gacgttcatg cttcttgtct tcttccacca aaaaagactc
    10621 ttcaagtcga tgaaagaaac tatcatctac tttcgccatc tcatttgaaa aaatctcctg
    10681 tagataacaa atacgattct tccgacgtgt ataccttcta cgagctgtcc gtttcagacg
    10741 agtcgcttcc gctgtctctc cactgtcaaa taaaagagcc cctataagat tttttttgat
    10801 actgtggcgg tctgtatttc ccagaacctt gaacttttta gacggaacct tatattcatc
    10861 agtgatcacc gcccatccga cgctatttgt gccgatatct aagcctattg agtatttctt
    10921 atccat
//
