LOCUS       sequence-371924-       10641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L-Rhamnose inducible catalytically inactive Cas9 with sgRNA for 
            bacterial gene knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10641)
  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 10641)
  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
FEATURES             Location/Qualifiers
     source          1..10641
                     /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"
     terminator      5139..5225
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      5352..5883
                     /label=oriV
                     /note="incP origin of replication"
     CDS             6276..7424
                     /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            7485..7593
                     /note="incP origin of transfer"
     CDS             complement(7760..8575)
                     /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     8567..8586
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(8707..8801)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(8861..9709)
                     /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(9783..10619)
                     /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 aaaggcatca aataaaacga aaggctcagt
     5161 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     5221 caaatccgcc gccctagaca gctgggcgcg ccgtagaaaa gatcaaagga tcttcttgag
     5281 atcctttttt tctgcggggg atcaggaccg ctgccggagc gcaacccact cactacagca
     5341 gagccatgta gggccgccgg cgttgtggat acctcgcgga aaacttggcc ctcactgaca
     5401 gatgaggggc ggacgttgac acttgagggg ccgactcacc cggcgcggcg ttgacagatg
     5461 aggggcaggc tcgatttcgg ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa
     5521 acgcctgatt ttacgcgagt ttcccacaga tgatgtggac aagcctgggg ataagtgccc
     5581 tgcggtattg acacttgagg ggcgcgacta ctgacagatg aggggcgcga tccttgacac
     5641 ttgaggggca gagtgctgac agatgagggg cgcacctatt gacatttgag gggctgtcca
     5701 caggcagaaa atccagcatt tgcaagggtt tccgcccgtt tttcggccac cgctaacctg
     5761 tcttttaacc tgcttttaaa ccaatattta taaaccttgt ttttaaccag ggctgcgccc
     5821 tgtgcgcgtg accgcgcacg ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc
     5881 gctaacgcgg gcctcccatc cccccagggg ctgcgcccct cggccgcgaa cggcctcacc
     5941 ccaaaaatgg cagccacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat
     6001 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct
     6061 tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg
     6121 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg
     6181 gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatcgacgga tcgatccggg
     6241 gaattaattc cggggcaatc ccgcaaggag ggtgaatgaa tcggacgttt gaccggaagg
     6301 catacaggca agaactgatc gacgcggggt tttccgccga ggatgccgaa accatcgcaa
     6361 gccgcaccgt catgcgtgcg ccccgcgaaa ccttccagtc cgtcggctcg atagtccagc
     6421 aagctacggc caagatcgag cgcgacagcg tgcaactggc tccccctgcc ctgcccgcgc
     6481 catcggccgc cgtggagcgt tcgcgtcgtc tcgaacagga ggcggcaggt ttggcgaagt
     6541 cgatgaccat cgacacgcga ggaactatga cgaccaagaa gcgaaaaacc gccggcgagg
     6601 acctggcaaa acaggtcagc gaagccaagc aggccgcgtt gctgaaacac acgaagcagc
     6661 agatcaagga aatgcagctt tccttgttcg atattgcgcc gtggccggac acgatgcgag
     6721 cgatgccaaa cgacacggcc cgctctgccc tgttcaccac gcgcaacaag aaaatcccgc
     6781 gcgaggcgct gcaaaacaag gtcattttcc acgtcaacaa ggacgtgaag atcacctaca
     6841 ccggcgtcga gctgcgggcc gacgatgacg aactggtgtg gcagcaggtg ttggagtacg
     6901 cgaagcgcac ccctatcggc gagccgatca ccttcacgtt ctacgagctt tgccaggacc
     6961 tgggctggtc gatcaatggc cggtattaca cgaaggccga ggaatgcctg tcgcgcctac
     7021 aggcgacggc gatgggcttc acgtccgacc gcgttgggca cctggaatcg gtgtcgctgc
     7081 tgcaccgctt ccgcgtcctg gaccgtggca agaaaacgtc ccgttgccag gtcctgatcg
     7141 acgaggaaat cgtcgtgctg tttgctggcg accactacac gaaattcata tgggagaagt
     7201 accgcaagct gtcgccgacg gcccgacgga tgttcgacta tttcagctcg caccgggagc
     7261 cgtacccgct caagctggaa accttccgcc tcatgtgcgg atcggattcc acccgcgtga
     7321 agaagtggcg cgagcaggtc ggcgaagcct gcgaagagtt gcgaggcagc ggcctggtgg
     7381 aacacgcctg ggtcaatgat gacctggtgc attgcaaacg ctagggcctt gtggggtcag
     7441 ttccggctgg gggttcagca gccacctgca ttgcggccgg cctacggcca gcctcgcaga
     7501 gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg
     7561 ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga
     7621 aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc
     7681 gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagga caacgcgcgg
     7741 accgcggtcc aattaattat tagaaaaatt catccagcat cagatgaaat tgcagtttgt
     7801 tcatatccgg attatcaatg ccatatttct gaaacagacg tttttgcagg ctcgggctaa
     7861 attcgcccag gcagttccac agaatggcca gatcctgata acgatccgca atgcccacac
     7921 ggcccacatc aatgcagcca atcagtttgc cttcatcgaa aatcaggtta tccaggctaa
     7981 aatcgccgtg ggtcaccacg ctatccgggc taaacggcag cagtttatgc atttctttcc
     8041 acacctgttc caccggccag ccgttacgtt catcatcaaa atcgctcgca tccaccaggc
     8101 cgttgttcat acggctctgc gcctgggcca gacgaaacac acgatcgctg ttaaacgggc
     8161 agttgcacac cggaatgcta tgcagacgac gcagaaacac ggccagcgca tccacaatgt
     8221 tttcgccgct atccggatat tcttccagca cctgaaacgc ggttttgccc ggaatcgcgg
     8281 tggtcagcag ccacgcatca tccggggtgc gaataaaatg tttaatggtc ggcagcggca
     8341 taaattcggt cagccagttc agacgcacca tttcatcggt cacatcgttc gccacgctgc
     8401 ctttgccatg tttcagaaac agttccggcg catccggttt gccatacaga cgataaatgg
     8461 tcgcgccgct ctgacccacg ttatcacgcg cccatttata gccatacaga tccgcatcca
     8521 tgttgctgtt cagacgcgga cggctacagc tcgtttcacg ctgaatatgg ctcataacac
     8581 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     8641 cttgtgcaat gtaacatcag agattttgag acacaaattt aaatcgtaat tattggggac
     8701 ccctggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     8761 atggagttct gaggtcatta ctggatctat caacaggagt ccaagactag tcgccagggt
     8821 tttcccagtc acgacgtttc tttaccagac tcgaggcgaa ttaatctttc tgcgaattga
     8881 gatgacgcca ctggctgggc gtcatcccgg tttcgcgggt aaacaccacc gaaaaatagt
     8941 tactatcttc aaagccacat tcggtcgaaa tatcactgat taacaggcgg ctatgctgga
     9001 gaagatattg cgcatgacac actctgacct gtcgcagata ttgattgatg gtcattccag
     9061 tctgctggcg aaattgctga cgcaaaacgc gctcactgca cgatgcctca tcacaaaatt
     9121 tatccagcgc aaagggactt ttcaggctag ccgccagccg ggtaatcagc ttatccagca
     9181 acgtttcgct ggatgttggc ggcaacgaat cactggtgta acgatggcga ttcagcaaca
     9241 tcaccaactg cccgaacagc aactcagcca tttcgttagc aaacggcaca tgctgactac
     9301 tttcatgctc aagctgaccg ataacctgcc gcgcctgcgc catccccatg ctacctaagc
     9361 gccagtgtgg ttgccctgcg ctggcgttaa atcccggaat cgccccctgc cagtcaagat
     9421 tcagcttcag acgctccggg caataaataa tattctgcaa aaccagatcg ttaacggaag
     9481 cgtaggagtg tttatcgtca gcatgaatgt aaaagagatc gccacgggta atgcgataag
     9541 ggcgatcgtt gagtacatgc aggccattac cgcgccagac aatcaccagc tcacaaaaat
     9601 catgtgtatg ttcagcaaag acatcttgcg gataacggtc agccacagcg actgcctgct
     9661 ggtcgctggc aaaaaaatca tctttgagaa gttttaactg atgcgccacc gtggctacct
     9721 cggccagaga acgaagttga ttattcgcaa tatggcgtac aaatacgttg agaagattcg
     9781 cgttattgca gaaagccatc ccgtccctgg cgaatatcac gcggtgacca gttaaactct
     9841 cggcgaaaaa gcgtcgaaaa gtggttactg tcgctgaatc cacagcgata ggcgatgtca
     9901 gtaacgctgg cctcgctgtg gcgtagcaga tgtcgggctt tcatcagtcg caggcggttc
     9961 aggtatcgct gaggcgtcag tcccgtttgc tgcttaagct gccgatgtag cgtacgcagt
    10021 gaaagagaaa attgatccgc cacggcatcc caattcacct catcggcaaa atggtcctcc
    10081 agccaggcca gaagcaagtt gagacgtgat gcgctgtttt ccaggttctc ctgcaaactg
    10141 cttttacgca gcaagagcag taattgcata aacaagatct cgcgactggc ggtcgagggt
    10201 aaatcatttt ccccttcctg ctgttccatc tgtgcaacca gctgtcgcac ctgctgcaat
    10261 acgctgtggt taacgcgcca gtgagacgga tactgcccat ccagctcttg tggcagcaac
    10321 tgattcagcc cggcgagaaa ctgaaatcga tccggcgagc gatacagcac attggtcaga
    10381 cacagattat cggtatgttc atacagatgc cgatcatgat cgcgtacgaa acagaccgtg
    10441 ccgccggtga tggtataggg ctgcccatta aacacatgaa tacccgtgcc atgttcgaca
    10501 atcacaattt catgaaaatc atgatgatgt tcaggaaaat ccgcctgcgg gagccggggt
    10561 tctatcgcca cggacgcgtt accagacgga aaaaaatcca cactatgtaa tacggtcata
    10621 ctggcctcct gatgtcgtca a
//
