LOCUS       sequence-357116-       11193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NT-CRISPR plasmid for a single gRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11193)
  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 11193)
  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_116_LVL2 cam
FEATURES             Location/Qualifiers
     source          1..11193
                     /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"
     protein_bind    1490..1508
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     RBS             1675..1686
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1693..2406
                     /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      2418..2489
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2505..2532
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2778..3366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2858..2877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3473..3507)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3578..4237)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4152..4171
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(5026..6108)
                     /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     6070..6089
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6185..6190,6191..6206,6207..6213)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6219..6238
                     /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             6247..6258
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(6919..6946)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6962..7033)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(7051..7083)
                     /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(7090..11193)
                     /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(7090..11193)
                     /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 ccgctgcttc aaatcactgt aatgatctta attcctaatt tttgttgaca ctctatcatt
     1501 gatagagtta ttttagtcca gagaccgaaa gtgaaacgtg atttcatgcg tcattttgaa
     1561 cattttgtaa atcttattta ataatgtgtg cggcaattca catttaattt atgaatgttt
     1621 tcttaacatc gcggcaactc aagaaacggc aggttcggat cttagctact agagaaagag
     1681 gagaaatact agatgcgtaa aggcgaagaa cttttcactg gcgtagtacc aattttagtg
     1741 gagctggacg gagatgtaaa tggtcataag ttttcagttc gaggagaagg cgaaggagat
     1801 gcaactaacg gtaagctgac actaaagttt atctgtacca cgggtaaact gccggtccca
     1861 tggccgacac tggttactac acttacttat ggtgtacaat gctttgctcg ttatcctgac
     1921 catatgaagc agcacgattt cttcaaatcg gccatgcctg aaggatacgt gcaagaacgt
     1981 acaattagct ttaaagatga cggcacctat aaaacgcgcg ccgaagttaa attcgagggt
     2041 gacacattgg ttaatcgtat agaacttaag ggcattgact tcaaagaaga tggcaacatc
     2101 cttggccata aactggaata taattttaac tctcacaatg tctacattac ggcggataaa
     2161 caaaagaatg gcattaaagc gaatttcaaa atccgccaca atgtcgaaga cggctcggtt
     2221 caactggcgg accattatca gcagaacacc ccaatcggtg atggcccggt tctgttacct
     2281 gataatcatt acctttctac tcaaagcgtt ttatctaaag atcctaacga gaagcgtgat
     2341 catatggttc tactggaatt tgttaccgca gctggtatca cgcacggcat ggatgagctg
     2401 tataaataac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2461 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     2521 ggcctttctg cgtttatagg tctcagtttc agagctatgc tggaaacagc atagcaagtt
     2581 gaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttctc
     2641 ggtaccaaat tccagaaaag aggcctcccg aaaggggggc cttttttcgt tttggtccac
     2701 cctgcacgat aacactgagg tagctgcgat tagacagcca gcagctgaac cgtaaaaaag
     2761 gccgcgttgc tggcgttttt ccacaggctc cgcccccctg acgagcatca caaaaatcga
     2821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcactgctac caataaaaaa
     3481 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     3541 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     3601 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     3661 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     3721 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     3781 aactcaccca gggattggct gaaacgaaaa acatattctc aataaaccct ttagggaaat
     3841 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     3901 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     3961 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     4021 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     4081 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     4141 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     4201 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     4261 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     4321 aacctcttac gtgcccgatc aaaacaggac caaaacgaaa aaagacgctt ttcagcgtct
     4381 tattgttcgt ctttggtacc gagctgtgtc tgtggctgag agagcaggta caagataaac
     4441 ccgtctatat tctcaaattc tttcactatc gcacttgttt tatgtgaagt ccagatatgg
     4501 actctggaac ccaactcgga gctctgcggg tagagcgtca accattcgaa cgttgcgcgg
     4561 tcttcggaga accgtaagag gtaatatact ccgatattca tttcagcggt agctgagtcc
     4621 gtcagttgat acggtgcgta cccggagcaa ataggagagg cggttgtagt ggggggaatg
     4681 gatagcaagc tgcgggtaga cctccaatta ttgaaggcct cccaaatcgg ggggcctttt
     4741 ttattgataa caaaaggagc tcagataaaa tatttgctca tgagcccgaa gtggcgagcc
     4801 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     4861 gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac agcttatcat
     4921 cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc
     4981 cctcgagccg tcaattgtct gattcgttac caattaggat ccttatcact gcccgctttc
     5041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5101 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagactg gcaacagctg
     5161 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5221 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctatcttc
     5281 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     5341 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     5401 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5461 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5521 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5581 tgcgaccaga tgctccacgc ccagtcgcgt accgtcctca tgggagtaaa taatactgtt
     5641 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5701 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5761 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5821 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5881 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5941 cagttgttgt gccacgcggt tgggaatgta attcagctcc accatcgccg cttccacttt
     6001 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtcatata
     6061 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcatat tcaccaccct
     6121 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcggc
     6181 tgtgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgc tcacaattta
     6241 ctagagaaag aggagaaata atcaatggat atgaatgagc aacagttttt cgactacgta
     6301 actaagtttg gcgcctacca aaaacgctcg atgtttggtg gtattggctt gtttcaacac
     6361 gacgctatgt atgtgttggt cagtgaagac cgcatttttg tgcgtggtgg agaagagctc
     6421 gaccctgagc tcttggcctt agggtgcgag aaatatcgcc atgttaaaaa acagacaacg
     6481 gcaaccgtaa actattacga catcactgaa ctgtatgagc aacatcaccc tgagctagac
     6541 tcgattattg aacgctcgat tcagttttct gtgaatcagc gggaatttca acgctcagca
     6601 gccagtcgtc gtctacggga tttacccaat atgcaactga ctttggagcg tatggtaaaa
     6661 aaagcgggca ttgacgatgt ggaaactttt atgagcctcg gtgcgccaga agtgtttaat
     6721 aaggtgcgtc aagcttatgg aagtgatgtc gatgtaaaac tactgtggaa gtttgcgggt
     6781 gcgattgaag gcattcactg gaaactgctg caagagccgc gtaagcgcca actgttagaa
     6841 agttgtcagc agcgtgctta aaaaaaaaaa ccccgcccct gacagggcgg ggtttttttt
     6901 cgcttactag cgtataaacg cagaaaggcc cacccgaagg tgagccagtg tgactctagt
     6961 agagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct
     7021 ttcgttttat ttgatgcctg gttaccccta agcagccaga gcgtagtttt cgtcgttagc
     7081 agctaaagcg tcacctccta gctgactcaa atcaatgcgt gtttcataaa gaccagtgat
     7141 ggattgatgg ataagagtgg catctaaaac ttcttttgta gacgtatatc gtttacgatc
     7201 aattgttgta tcaaaatatt taaaagcagc gggagctcca agattcgtca acgtaaataa
     7261 atgaataata ttttctgctt gttcacgtat tggtttgtct ctatgtttgt tatatgcact
     7321 aagaacttta tctaaattgg catctgctaa aataacacgc ttagaaaatt cactgatttg
     7381 ctcaataatc tcatctaaat aatgcttatg ctgctccaca aacaattgtt tttgttcgtt
     7441 atcttctgga ctacccttca acttttcata atgactagct aaatataaaa aattcacata
     7501 tttgcttggc agagccagct catttccttt ttgtaattct ccggcactag ccagcatccg
     7561 tttacgaccg ttttctaact caaaaagact atatttaggt agtttaatga ttaagtcttt
     7621 tttaacttcc ttatatcctt tagcttctaa aaagtcaatc ggattttttt caaaggaact
     7681 tctttccata attgtgatcc ctagtaactc tttaacggat tttaacttct tcgatttccc
     7741 tttttccacc ttagcaacca ctaggactga ataagctacc gttggactat caaaaccacc
     7801 atattttttt ggatcccagt cttttttacg agcaataagc ttgtccgaat ttctttttgg
     7861 taaaattgac tccttggaga atccgcctgt ctgtacttct gttttcttga caatattgac
     7921 ttggggcatg gacaatactt tgcgcactgt ggcaaaatct cgccctttat cccagacaat
     7981 ttctccagtt tccccattag tttcgattag agggcgtttg cgaatctctc catttgcaag
     8041 tgtaatttct gttttgaaga agttcatgat attagagtaa aagaaatatt ttgcggttgc
     8101 tttgcctatt tcttgctcag acttagcaat cattttacga acatcataaa ctttataatc
     8161 accatagaca aactccgatt caagttttgg atatttctta atcaaagcag ttccaacgac
     8221 ggcatttaga tacgcatcat gggcatgatg gtaattgtta atctcacgta ctttatagaa
     8281 ttggaaatct tttcggaagt cagaaactaa tttagatttt aaggtaatca ctttaacctc
     8341 tcgaataagt ttatcatttt catcgtattt agtattcatg cgactatcca aaatttgtgc
     8401 cacatgctta gtgatttggc gagtttcaac caattggcgt ttgataaaac cagctttatc
     8461 aagttcactc aaacctccac gttcagcttt cgttaaatta tcaaacttac gttgagtgat
     8521 taacttggcg tttagaagtt gtctccaata gtttttcatc tttttgacta cttcttcact
     8581 tggaacgtta tccgatttac cacgattttt atcagaacgc gttaagacct tattgtctat
     8641 tgaatcgtct ttaaggaaac tttgtggaac aatgtgatcg acatcataat cacttaaacg
     8701 attaatatct aattcttggt ccacatacat gtctcttcca ttttggagat aatagagata
     8761 gagcttttca ttttgcaatt gagtattttc aacaggatgc tctttaagaa tctgacttcc
     8821 taattctttg ataccttctt cgattcgttt catacgctct cgcgaatttt tctggccctt
     8881 ttgagttgtc tgattttcac gtgccatttc aataacgata ttttctggct tatgccgccc
     8941 cattactttg accaattcat caacaacttt tacagtctgt aaaatacctt ttttaatagc
     9001 agggctacca gctaaatttg caatatgttc atgtaaacta tcgccttgtc cagacacttg
     9061 tgctttttga atgtcttctt taaatgtcaa actatcatca tggatcagct gcataaaatt
     9121 gcgattggca aaaccatctg atttcaaaaa atctaatatt gttttgccag attgcttatc
     9181 cctaatacca ttaatcaatt ttcgagacaa acgtccccaa ccagtataac ggcgacgttt
     9241 aagctgtttc atcaccttat catcaaagag gtgagcatat gttttaagtc tttcctcaat
     9301 catctcccta tcttcaaata aggtcaatgt taaaacaata tcctctaaga tatcttcatt
     9361 ttcttcatta tccaaaaaat ctttatcttt aataattttt agcaaatcat ggtaggtacc
     9421 taatgaagca ttaaatctat cttcaactcc tgaaatttca acactatcaa aacattctat
     9481 ttttttgaaa taatcttctt ttaattgctt aacggttact tttcgatttg ttttgaagag
     9541 taaatcaaca atggctttct tctgttcacc tgaaagaaat gctggttttc gcattccttc
     9601 agtaacatat ttgacctttg tcaattcgtt ataaaccgta aaatactcat aaagcaaact
     9661 atgttttggt agtacttttt catttggaag atttttatca aagtttgtca tgcgttcaat
     9721 aaatgattga gctgaagcac ctttatcgac aacttcttca aaattccatg gggtaattgt
     9781 ttcttcagac ttccgagtca tccatgcaaa acgactattg ccacgcgcca atggaccaac
     9841 ataataagga attcgaaaag tcaagatttt ttcaatcttc tcacgattgt cttttaaaaa
     9901 tggataaaag tcttcttgtc ttctcaaaat agcatgcagc tcacccaagt gaatttgatg
     9961 gggaatagag ccgttgtcaa aggtccgttg cttgcgcagc aaatcttcac gatttagttt
    10021 caccaataat tcctcagtac catccatttt ttctaaaatt ggtttgataa atttataaaa
    10081 ttcttcttgg ctagctcccc catcaatata acctgcatat ccgttttttg attgatcaaa
    10141 aaagatttct ttatactttt ctggaagttg ttgtcgaact aaagctttta aaagagtcaa
    10201 gtcttgatga tgttcatcgt agcgtttaat cattgaagct gataggggag ccttagttat
    10261 ttcagtattt actcttagga tatctgaaag taaaatagca tctgataaat tcttagctgc
    10321 caaaaacaaa tcagcatatt gatctccaat ttgcgccaat aaattatcta aatcatcatc
    10381 gtaagtatct tttgaaagct gtaatttagc atcttctgcc aaatcaaaat ttgatttaaa
    10441 attaggggtc aaacccaatg acaaagcaat gagattccca aataagccat ttttcttctc
    10501 accggggagc tgagcaatga gattttctaa tcgtcttgat ttactcaatc gtgcagaaag
    10561 aatcgcttta gcatctactc cacttgcgtt aatagggttt tcttcaaata attgattgta
    10621 ggtttgtacc aactggataa atagtttgtc cacatcacta ttatcaggat ttaaatctcc
    10681 ctcaatcaaa aaatgaccac gaaacttaat catatgcgct aaggccaaat agattaagcg
    10741 caaatccgct ttatcagtag aatctaccaa tttttttcgc agatgataga tagttggata
    10801 tttctcatga taagcaactt catctactat atttccaaaa ataggatgac gttcatgctt
    10861 cttgtcttct tccaccaaaa aagactcttc aagtcgatga aagaaactat catctacttt
    10921 cgccatctca tttgaaaaaa tctcctgtag ataacaaata cgattcttcc gacgtgtata
    10981 ccttctacga gctgtccgtt tcagacgagt cgcttccgct gtctctccac tgtcaaataa
    11041 aagagcccct ataagatttt ttttgatact gtggcggtct gtatttccca gaaccttgaa
    11101 ctttttagac ggaaccttat attcatcagt gatcaccgcc catccgacgc tatttgtgcc
    11161 gatatctaag cctattgagt atttcttatc cat
//
