LOCUS       sequence-371637-       11563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L-Rhamnose inducible catalytically inactive Cas9 with sgRNA and racE
            gene for bacterial gene knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11563)
  AUTHORS   Kim SK, Kim H, Woo SG, Kim TH, Rha E, Kwon KK, Lee H, Lee SG, Lee DH
  TITLE     CRISPRi-based programmable logic inverter cascade for 
            antibiotic-free selection and maintenance of multiple plasmids.
  JOURNAL   Nucleic Acids Res. 2022 Dec 9;50(22):13155-13171. doi: 
            10.1093/nar/gkac1104.
  PUBMED    36511859
REFERENCE   2  (bases 1 to 11563)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2022-12-9- 9"; volume: "50"; issue: 
            "22"; pages: "13155-13171"
COMMENT     Sequence Label: pSLiP-G2-RacE
FEATURES             Location/Qualifiers
     source          1..11563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        124..242
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             join(266..292,293..295,296..2782,2783..2785,2786..4372)
                     /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
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     terminator      4396..4467
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4483..4510
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4560..4594
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        4615..4690
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             4733..4762
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4778..4795
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4851..4868)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5021..5067
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        5133..5167
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     terminator      6061..6147
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      6274..6805
                     /label=oriV
                     /note="incP origin of replication"
     CDS             7198..8346
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     oriT            8407..8515
                     /note="incP origin of transfer"
     CDS             complement(8682..9497)
                     /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"
     primer_bind     9489..9508
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(9629..9723)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(9783..10631)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(10705..11541)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
ORIGIN
        1 cacggcgaaa tagtaatcac gaggtcaggt tcttacctta aattttcgac ggaaaaccac
       61 gtaaaaaacg tcgatttttc aagatacagc gtgaattttc aggaaatgcg gtgagcatca
      121 catcaccaca attcagcaaa ttgtgaacat catcacgttc atctttccct ggttgccaat
      181 ggcccatttt cctgtcagta acgagaaggt cgcgaattca ggcgcttttt agactggtcg
      241 taatgaaatt cagcaggatc accatatgga taagaaatac tcaataggct tagctatcgg
      301 cacaaatagc gtcggatggg cggtgatcac tgatgaatat aaggttccgt ctaaaaagtt
      361 caaggttctg ggaaatacag accgccacag tatcaaaaaa aatcttatag gggctctttt
      421 atttgacagt ggagagacag cggaagcgac tcgtctcaaa cggacagctc gtagaaggta
      481 tacacgtcgg aagaatcgta tttgttatct acaggagatt ttttcaaatg agatggcgaa
      541 agtagatgat agtttctttc atcgacttga agagtctttt ttggtggaag aagacaagaa
      601 gcatgaacgt catcctattt ttggaaatat agtagatgaa gttgcttatc atgagaaata
      661 tccaactatc tatcatctgc gaaaaaaatt ggtagattct actgataaag cggatttgcg
      721 cttaatctat ttggccttag cgcatatgat taagtttcgt ggtcattttt tgattgaggg
      781 agatttaaat cctgataata gtgatgtgga caaactattt atccagttgg tacaaaccta
      841 caatcaatta tttgaagaaa accctattaa cgcaagtgga gtagatgcta aagcgattct
      901 ttctgcacga ttgagtaaat caagacgatt agaaaatctc attgctcagc tccccggtga
      961 gaagaaaaat ggcttatttg ggaatctcat tgctttgtca ttgggtttga cccctaattt
     1021 taaatcaaat tttgatttgg cagaagatgc taaattacag ctttcaaaag atacttacga
     1081 tgatgattta gataatttat tggcgcaaat tggagatcaa tatgctgatt tgtttttggc
     1141 agctaagaat ttatcagatg ctattttact ttcagatatc ctaagagtaa atactgaaat
     1201 aactaaggct cccctatcag cttcaatgat taaacgctac gatgaacatc atcaagactt
     1261 gactctttta aaagctttag ttcgacaaca acttccagaa aagtataaag aaatcttttt
     1321 tgatcaatca aaaaacggat atgcaggtta tattgatggg ggagctagcc aagaagaatt
     1381 ttataaattt atcaaaccaa ttttagaaaa aatggatggt actgaggaat tattggtgaa
     1441 actaaatcgt gaagatttgc tgcgcaagca acggaccttt gacaacggct ctattcccca
     1501 tcaaattcac ttgggtgagc tgcatgctat tttgagaaga caagaagact tttatccatt
     1561 tttaaaagac aatcgtgaga agattgaaaa aatcttgact tttcgaattc cttattatgt
     1621 tggtccattg gcgcgtggca atagtcgttt tgcatggatg actcggaagt ctgaagaaac
     1681 aattacccca tggaattttg aagaagttgt cgataaaggt gcttcagctc aatcatttat
     1741 tgaacgcatg acaaactttg ataaaaatct tccaaatgaa aaagtactac caaaacatag
     1801 tttgctttat gagtatttta cggtttataa cgaattgaca aaggtcaaat atgttactga
     1861 aggaatgcga aaaccagcat ttctttcagg tgaacagaag aaagccattg ttgatttact
     1921 cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa gaagattatt tcaaaaaaat
     1981 agaatgtttt gatagtgttg aaatttcagg agttgaagat agatttaatg cttcattagg
     2041 tacctaccat gatttgctaa aaattattaa agataaagat tttttggata atgaagaaaa
     2101 tgaagatatc ttagaggata ttgttttaac attgacctta tttgaagata gggagatgat
     2161 tgaggaaaga cttaaaacat atgctcacct ctttgatgat aaggtgatga aacagcttaa
     2221 acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa ttgattaatg gtattaggga
     2281 taagcaatct ggcaaaacaa tattagattt tttgaaatca gatggttttg ccaatcgcaa
     2341 ttttatgcag ctgatccatg atgatagttt gacatttaaa gaagacattc aaaaagcaca
     2401 agtgtctgga caaggcgata gtttacatga acatattgca aatttagctg gtagccctgc
     2461 tattaaaaaa ggtattttac agactgtaaa agttgttgat gaattggtca aagtaatggg
     2521 gcggcataag ccagaaaata tcgttattga aatggcacgt gaaaatcaga caactcaaaa
     2581 gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca aagaattagg
     2641 aagtcagatt cttaaagagc atcctgttga aaatactcaa ttgcaaaatg aaaagctcta
     2701 tctctattat ctccaaaatg gaagagacat gtatgtggac caagaattag atattaatcg
     2761 tttaagtgat tatgatgtcg atgccattgt tccacaaagt ttccttaaag acgattcaat
     2821 agacaataag gtcttaacgc gttctgataa aaatcgtggt aaatcggata acgttccaag
     2881 tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg ccaagttaat
     2941 cactcaacgt aagtttgata atttaacgaa agctgaacgt ggaggtttga gtgaacttga
     3001 taaagctggt tttatcaaac gccaattggt tgaaactcgc caaatcacta agcatgtggc
     3061 acaaattttg gatagtcgca tgaatactaa atacgatgaa aatgataaac ttattcgaga
     3121 ggttaaagtg attaccttaa aatctaaatt agtttctgac ttccgaaaag atttccaatt
     3181 ctataaagta cgtgagatta acaattacca tcatgcccat gatgcgtatc taaatgccgt
     3241 cgttggaact gctttgatta agaaatatcc aaaacttgaa tcggagtttg tctatggtga
     3301 ttataaagtt tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa taggcaaagc
     3361 aaccgcaaaa tatttctttt actctaatat catgaacttc ttcaaaacag aaattacact
     3421 tgcaaatgga gagattcgca aacgccctct aatcgaaact aatggggaaa ctggagaaat
     3481 tgtctgggat aaagggcgag attttgccac agtgcgcaaa gtattgtcca tgccccaagt
     3541 caatattgtc aagaaaacag aagtacagac aggcggattc tccaaggagt caattttacc
     3601 aaaaagaaat tcggacaagc ttattgctcg taaaaaagac tgggatccaa aaaaatatgg
     3661 tggttttgat agtccaacgg tagcttattc agtcctagtg gttgctaagg tggaaaaagg
     3721 gaaatcgaag aagttaaaat ccgttaaaga gttactaggg atcacaatta tggaaagaag
     3781 ttcctttgaa aaaaatccga ttgacttttt agaagctaaa ggatataagg aagttaaaaa
     3841 agacttaatc attaaactac ctaaatatag tctttttgag ttagaaaacg gtcgtaaacg
     3901 gatgctggct agtgccggag aattacaaaa aggaaatgag ctggctctgc caagcaaata
     3961 tgtgaatttt ttatatttag ctagtcatta tgaaaagttg aagggtagtc cagaagataa
     4021 cgaacaaaaa caattgtttg tggagcagca taagcattat ttagatgaga ttattgagca
     4081 aatcagtgaa ttttctaagc gtgttatttt agcagatgcc aatttagata aagttcttag
     4141 tgcatataac aaacatagag acaaaccaat acgtgaacaa gcagaaaata ttattcattt
     4201 atttacgttg acgaatcttg gagctcccgc tgcttttaaa tattttgata caacaattga
     4261 tcgtaaacga tatacgtcta caaaagaagt tttagatgcc actcttatcc atcaatccat
     4321 cactggtctt tatgaaacac gcattgattt gagtcagcta ggaggtgact aactcgagta
     4381 aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     4441 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     4501 cctttctgcg tttataccta gggatatatt ccgcttcctc ggcgaccggt taaagatctt
     4561 tgacagctag ctcagtccta ggtataatcc tagtccatgt aatcagcgtc taacgtttta
     4621 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     4681 gagtcggtgc tttttttgac ccggggcggc cgcctcgagg aagcttgggc ccgaacaaaa
     4741 actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt
     4801 ttaaacggtc tccagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag
     4861 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     4921 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     4981 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     5041 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaactggc gcccaatacg
     5101 caaaccgtga aattgttatc cgctttaatt aatttatggc tagctcagtc ctaggtacaa
     5161 tgctagcatc cgccaacctg gagaaggaga tatacatatg ttggaacaac caataggagt
     5221 cattgattcc ggggttggcg gtttaaccgt tgcgaaggaa atcatgagac agctacctaa
     5281 agaaaatatt atctacgtcg gggatacgaa acggtgtcct tacgggccgc gccctgaaga
     5341 agaggtgctt caatatacgt gggagctgac gaattattta ctcgaaaacc accacatcaa
     5401 aatgctcgtg atcgcctgta atacagcaac agcgatcgct ttggatgaca tccagcgcag
     5461 cgtcggtata ccggtggtcg gagtcatcca gcctggtgcg agagcagcga taaaagtgac
     5521 ggataatcag catatcggtg tcatcggcac agagaatacg attaagagca atgcatacga
     5581 agaagcgctt ttggcattaa accctgattt gaaggttgaa aaccttgcct gcccgctgct
     5641 tgtgcctttt gtggaaagcg ggaagtttct cgacaaaaca gcagacgaga ttgttaaaac
     5701 ctcgctgtat ccgttaaaag acacatcaat tgattcgctg attttaggct gcacccatta
     5761 ccctatttta aaagaagcca ttcaaagata tatgggagag cacgtaaaca ttatttcgtc
     5821 cggcgatgaa acagcccggg aagtcagcac aattctctct tataaagggc tgctgaacca
     5881 gtctccgatt gccccggatc atcagttcct gacaacaggg gcgcgtgatc agtttgcaaa
     5941 aatcgcagac gattggtttg gccatgaagt cgggcatgtg gaatgtatct cactgcaaga
     6001 accgattaaa agagcagcaa acgacgaaaa ctacgcttta gcagcttaaa ttaaaggcat
     6061 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     6121 gtgaacgctc tcctgagtag gacaaatccg ccgccctaga cagctgggcg cgccgtagaa
     6181 aagatcaaag gatcttcttg agatcctttt tttctgcggg ggatcaggac cgctgccgga
     6241 gcgcaaccca ctcactacag cagagccatg tagggccgcc ggcgttgtgg atacctcgcg
     6301 gaaaacttgg ccctcactga cagatgaggg gcggacgttg acacttgagg ggccgactca
     6361 cccggcgcgg cgttgacaga tgaggggcag gctcgatttc ggccggcgac gtggagctgg
     6421 ccagcctcgc aaatcggcga aaacgcctga ttttacgcga gtttcccaca gatgatgtgg
     6481 acaagcctgg ggataagtgc cctgcggtat tgacacttga ggggcgcgac tactgacaga
     6541 tgaggggcgc gatccttgac acttgagggg cagagtgctg acagatgagg ggcgcaccta
     6601 ttgacatttg aggggctgtc cacaggcaga aaatccagca tttgcaaggg tttccgcccg
     6661 tttttcggcc accgctaacc tgtcttttaa cctgctttta aaccaatatt tataaacctt
     6721 gtttttaacc agggctgcgc cctgtgcgcg tgaccgcgca cgccgaaggg gggtgccccc
     6781 ccttctcgaa ccctcccggc ccgctaacgc gggcctccca tccccccagg ggctgcgccc
     6841 ctcggccgcg aacggcctca ccccaaaaat ggcagccacg tagaaagcca gtccgcagaa
     6901 acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
     6961 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
     7021 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
     7081 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
     7141 aagatcgacg gatcgatccg gggaattaat tccggggcaa tcccgcaagg agggtgaatg
     7201 aatcggacgt ttgaccggaa ggcatacagg caagaactga tcgacgcggg gttttccgcc
     7261 gaggatgccg aaaccatcgc aagccgcacc gtcatgcgtg cgccccgcga aaccttccag
     7321 tccgtcggct cgatagtcca gcaagctacg gccaagatcg agcgcgacag cgtgcaactg
     7381 gctccccctg ccctgcccgc gccatcggcc gccgtggagc gttcgcgtcg tctcgaacag
     7441 gaggcggcag gtttggcgaa gtcgatgacc atcgacacgc gaggaactat gacgaccaag
     7501 aagcgaaaaa ccgccggcga ggacctggca aaacaggtca gcgaagccaa gcaggccgcg
     7561 ttgctgaaac acacgaagca gcagatcaag gaaatgcagc tttccttgtt cgatattgcg
     7621 ccgtggccgg acacgatgcg agcgatgcca aacgacacgg cccgctctgc cctgttcacc
     7681 acgcgcaaca agaaaatccc gcgcgaggcg ctgcaaaaca aggtcatttt ccacgtcaac
     7741 aaggacgtga agatcaccta caccggcgtc gagctgcggg ccgacgatga cgaactggtg
     7801 tggcagcagg tgttggagta cgcgaagcgc acccctatcg gcgagccgat caccttcacg
     7861 ttctacgagc tttgccagga cctgggctgg tcgatcaatg gccggtatta cacgaaggcc
     7921 gaggaatgcc tgtcgcgcct acaggcgacg gcgatgggct tcacgtccga ccgcgttggg
     7981 cacctggaat cggtgtcgct gctgcaccgc ttccgcgtcc tggaccgtgg caagaaaacg
     8041 tcccgttgcc aggtcctgat cgacgaggaa atcgtcgtgc tgtttgctgg cgaccactac
     8101 acgaaattca tatgggagaa gtaccgcaag ctgtcgccga cggcccgacg gatgttcgac
     8161 tatttcagct cgcaccggga gccgtacccg ctcaagctgg aaaccttccg cctcatgtgc
     8221 ggatcggatt ccacccgcgt gaagaagtgg cgcgagcagg tcggcgaagc ctgcgaagag
     8281 ttgcgaggca gcggcctggt ggaacacgcc tgggtcaatg atgacctggt gcattgcaaa
     8341 cgctagggcc ttgtggggtc agttccggct gggggttcag cagccacctg cattgcggcc
     8401 ggcctacggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     8461 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     8521 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     8581 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc
     8641 agcggaaaag gacaacgcgc ggaccgcggt ccaattaatt attagaaaaa ttcatccagc
     8701 atcagatgaa attgcagttt gttcatatcc ggattatcaa tgccatattt ctgaaacaga
     8761 cgtttttgca ggctcgggct aaattcgccc aggcagttcc acagaatggc cagatcctga
     8821 taacgatccg caatgcccac acggcccaca tcaatgcagc caatcagttt gccttcatcg
     8881 aaaatcaggt tatccaggct aaaatcgccg tgggtcacca cgctatccgg gctaaacggc
     8941 agcagtttat gcatttcttt ccacacctgt tccaccggcc agccgttacg ttcatcatca
     9001 aaatcgctcg catccaccag gccgttgttc atacggctct gcgcctgggc cagacgaaac
     9061 acacgatcgc tgttaaacgg gcagttgcac accggaatgc tatgcagacg acgcagaaac
     9121 acggccagcg catccacaat gttttcgccg ctatccggat attcttccag cacctgaaac
     9181 gcggttttgc ccggaatcgc ggtggtcagc agccacgcat catccggggt gcgaataaaa
     9241 tgtttaatgg tcggcagcgg cataaattcg gtcagccagt tcagacgcac catttcatcg
     9301 gtcacatcgt tcgccacgct gcctttgcca tgtttcagaa acagttccgg cgcatccggt
     9361 ttgccataca gacgataaat ggtcgcgccg ctctgaccca cgttatcacg cgcccattta
     9421 tagccataca gatccgcatc catgttgctg ttcagacgcg gacggctaca gctcgtttca
     9481 cgctgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     9541 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaaat
     9601 ttaaatcgta attattgggg acccctggat tctcaccaat aaaaaacgcc cggcggcaac
     9661 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     9721 gtccaagact agtcgccagg gttttcccag tcacgacgtt tctttaccag actcgaggcg
     9781 aattaatctt tctgcgaatt gagatgacgc cactggctgg gcgtcatccc ggtttcgcgg
     9841 gtaaacacca ccgaaaaata gttactatct tcaaagccac attcggtcga aatatcactg
     9901 attaacaggc ggctatgctg gagaagatat tgcgcatgac acactctgac ctgtcgcaga
     9961 tattgattga tggtcattcc agtctgctgg cgaaattgct gacgcaaaac gcgctcactg
    10021 cacgatgcct catcacaaaa tttatccagc gcaaagggac ttttcaggct agccgccagc
    10081 cgggtaatca gcttatccag caacgtttcg ctggatgttg gcggcaacga atcactggtg
    10141 taacgatggc gattcagcaa catcaccaac tgcccgaaca gcaactcagc catttcgtta
    10201 gcaaacggca catgctgact actttcatgc tcaagctgac cgataacctg ccgcgcctgc
    10261 gccatcccca tgctacctaa gcgccagtgt ggttgccctg cgctggcgtt aaatcccgga
    10321 atcgccccct gccagtcaag attcagcttc agacgctccg ggcaataaat aatattctgc
    10381 aaaaccagat cgttaacgga agcgtaggag tgtttatcgt cagcatgaat gtaaaagaga
    10441 tcgccacggg taatgcgata agggcgatcg ttgagtacat gcaggccatt accgcgccag
    10501 acaatcacca gctcacaaaa atcatgtgta tgttcagcaa agacatcttg cggataacgg
    10561 tcagccacag cgactgcctg ctggtcgctg gcaaaaaaat catctttgag aagttttaac
    10621 tgatgcgcca ccgtggctac ctcggccaga gaacgaagtt gattattcgc aatatggcgt
    10681 acaaatacgt tgagaagatt cgcgttattg cagaaagcca tcccgtccct ggcgaatatc
    10741 acgcggtgac cagttaaact ctcggcgaaa aagcgtcgaa aagtggttac tgtcgctgaa
    10801 tccacagcga taggcgatgt cagtaacgct ggcctcgctg tggcgtagca gatgtcgggc
    10861 tttcatcagt cgcaggcggt tcaggtatcg ctgaggcgtc agtcccgttt gctgcttaag
    10921 ctgccgatgt agcgtacgca gtgaaagaga aaattgatcc gccacggcat cccaattcac
    10981 ctcatcggca aaatggtcct ccagccaggc cagaagcaag ttgagacgtg atgcgctgtt
    11041 ttccaggttc tcctgcaaac tgcttttacg cagcaagagc agtaattgca taaacaagat
    11101 ctcgcgactg gcggtcgagg gtaaatcatt ttccccttcc tgctgttcca tctgtgcaac
    11161 cagctgtcgc acctgctgca atacgctgtg gttaacgcgc cagtgagacg gatactgccc
    11221 atccagctct tgtggcagca actgattcag cccggcgaga aactgaaatc gatccggcga
    11281 gcgatacagc acattggtca gacacagatt atcggtatgt tcatacagat gccgatcatg
    11341 atcgcgtacg aaacagaccg tgccgccggt gatggtatag ggctgcccat taaacacatg
    11401 aatacccgtg ccatgttcga caatcacaat ttcatgaaaa tcatgatgat gttcaggaaa
    11461 atccgcctgc gggagccggg gttctatcgc cacggacgcg ttaccagacg gaaaaaaatc
    11521 cacactatgt aatacggtca tactggcctc ctgatgtcgt caa
//
