LOCUS       sequence-374348-       10796 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 10796)
  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 10796)
  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-G1G2
FEATURES             Location/Qualifiers
     source          1..10796
                     /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"
     promoter        4715..4749
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        4770..4845
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             4888..4917
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4933..4950
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5006..5023)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5176..5222
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5294..5380
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      5507..6038
                     /label=oriV
                     /note="incP origin of replication"
     CDS             6431..7579
                     /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            7640..7748
                     /note="incP origin of transfer"
     CDS             complement(7915..8730)
                     /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     8722..8741
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(8862..8956)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(9016..9864)
                     /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(9938..10774)
                     /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 tagtggaaaa caatgcgacc gatagtttta
     4621 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     4681 gagtcggtgc tttttttgac ccggttaaag atctttgaca gctagctcag tcctaggtat
     4741 aatcctagtc catgtaatca gcgtctaacg ttttagagct agaaatagca agttaaaata
     4801 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgacccggg
     4861 gcggccgcct cgaggaagct tgggcccgaa caaaaactca tctcagaaga ggatctgaat
     4921 agcgccgtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
     4981 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     5041 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     5101 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     5161 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     5221 atctgttgtt tgtcggtgaa ctggcgccca atacgcaaac cgtgaaattg ttatccgctt
     5281 taattaaagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     5341 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct agacagctgg
     5401 gcgcgccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gggggatcag
     5461 gaccgctgcc ggagcgcaac ccactcacta cagcagagcc atgtagggcc gccggcgttg
     5521 tggatacctc gcggaaaact tggccctcac tgacagatga ggggcggacg ttgacacttg
     5581 aggggccgac tcacccggcg cggcgttgac agatgagggg caggctcgat ttcggccggc
     5641 gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc
     5701 acagatgatg tggacaagcc tggggataag tgccctgcgg tattgacact tgaggggcgc
     5761 gactactgac agatgagggg cgcgatcctt gacacttgag gggcagagtg ctgacagatg
     5821 aggggcgcac ctattgacat ttgaggggct gtccacaggc agaaaatcca gcatttgcaa
     5881 gggtttccgc ccgtttttcg gccaccgcta acctgtcttt taacctgctt ttaaaccaat
     5941 atttataaac cttgttttta accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa
     6001 ggggggtgcc cccccttctc gaaccctccc ggcccgctaa cgcgggcctc ccatcccccc
     6061 aggggctgcg cccctcggcc gcgaacggcc tcaccccaaa aatggcagcc acgtagaaag
     6121 ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa
     6181 gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc
     6241 tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg
     6301 gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat
     6361 ggcgcagggg atcaagatcg acggatcgat ccggggaatt aattccgggg caatcccgca
     6421 aggagggtga atgaatcgga cgtttgaccg gaaggcatac aggcaagaac tgatcgacgc
     6481 ggggttttcc gccgaggatg ccgaaaccat cgcaagccgc accgtcatgc gtgcgccccg
     6541 cgaaaccttc cagtccgtcg gctcgatagt ccagcaagct acggccaaga tcgagcgcga
     6601 cagcgtgcaa ctggctcccc ctgccctgcc cgcgccatcg gccgccgtgg agcgttcgcg
     6661 tcgtctcgaa caggaggcgg caggtttggc gaagtcgatg accatcgaca cgcgaggaac
     6721 tatgacgacc aagaagcgaa aaaccgccgg cgaggacctg gcaaaacagg tcagcgaagc
     6781 caagcaggcc gcgttgctga aacacacgaa gcagcagatc aaggaaatgc agctttcctt
     6841 gttcgatatt gcgccgtggc cggacacgat gcgagcgatg ccaaacgaca cggcccgctc
     6901 tgccctgttc accacgcgca acaagaaaat cccgcgcgag gcgctgcaaa acaaggtcat
     6961 tttccacgtc aacaaggacg tgaagatcac ctacaccggc gtcgagctgc gggccgacga
     7021 tgacgaactg gtgtggcagc aggtgttgga gtacgcgaag cgcaccccta tcggcgagcc
     7081 gatcaccttc acgttctacg agctttgcca ggacctgggc tggtcgatca atggccggta
     7141 ttacacgaag gccgaggaat gcctgtcgcg cctacaggcg acggcgatgg gcttcacgtc
     7201 cgaccgcgtt gggcacctgg aatcggtgtc gctgctgcac cgcttccgcg tcctggaccg
     7261 tggcaagaaa acgtcccgtt gccaggtcct gatcgacgag gaaatcgtcg tgctgtttgc
     7321 tggcgaccac tacacgaaat tcatatggga gaagtaccgc aagctgtcgc cgacggcccg
     7381 acggatgttc gactatttca gctcgcaccg ggagccgtac ccgctcaagc tggaaacctt
     7441 ccgcctcatg tgcggatcgg attccacccg cgtgaagaag tggcgcgagc aggtcggcga
     7501 agcctgcgaa gagttgcgag gcagcggcct ggtggaacac gcctgggtca atgatgacct
     7561 ggtgcattgc aaacgctagg gccttgtggg gtcagttccg gctgggggtt cagcagccac
     7621 ctgcattgcg gccggcctac ggccagcctc gcagagcagg attcccgttg agcaccgcca
     7681 ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta
     7741 tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa
     7801 atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga
     7861 agcagggtta tgcagcggaa aaggacaacg cgcggaccgc ggtccaatta attattagaa
     7921 aaattcatcc agcatcagat gaaattgcag tttgttcata tccggattat caatgccata
     7981 tttctgaaac agacgttttt gcaggctcgg gctaaattcg cccaggcagt tccacagaat
     8041 ggccagatcc tgataacgat ccgcaatgcc cacacggccc acatcaatgc agccaatcag
     8101 tttgccttca tcgaaaatca ggttatccag gctaaaatcg ccgtgggtca ccacgctatc
     8161 cgggctaaac ggcagcagtt tatgcatttc tttccacacc tgttccaccg gccagccgtt
     8221 acgttcatca tcaaaatcgc tcgcatccac caggccgttg ttcatacggc tctgcgcctg
     8281 ggccagacga aacacacgat cgctgttaaa cgggcagttg cacaccggaa tgctatgcag
     8341 acgacgcaga aacacggcca gcgcatccac aatgttttcg ccgctatccg gatattcttc
     8401 cagcacctga aacgcggttt tgcccggaat cgcggtggtc agcagccacg catcatccgg
     8461 ggtgcgaata aaatgtttaa tggtcggcag cggcataaat tcggtcagcc agttcagacg
     8521 caccatttca tcggtcacat cgttcgccac gctgcctttg ccatgtttca gaaacagttc
     8581 cggcgcatcc ggtttgccat acagacgata aatggtcgcg ccgctctgac ccacgttatc
     8641 acgcgcccat ttatagccat acagatccgc atccatgttg ctgttcagac gcggacggct
     8701 acagctcgtt tcacgctgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     8761 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     8821 ttgagacaca aatttaaatc gtaattattg gggacccctg gattctcacc aataaaaaac
     8881 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
     8941 tctatcaaca ggagtccaag actagtcgcc agggttttcc cagtcacgac gtttctttac
     9001 cagactcgag gcgaattaat ctttctgcga attgagatga cgccactggc tgggcgtcat
     9061 cccggtttcg cgggtaaaca ccaccgaaaa atagttacta tcttcaaagc cacattcggt
     9121 cgaaatatca ctgattaaca ggcggctatg ctggagaaga tattgcgcat gacacactct
     9181 gacctgtcgc agatattgat tgatggtcat tccagtctgc tggcgaaatt gctgacgcaa
     9241 aacgcgctca ctgcacgatg cctcatcaca aaatttatcc agcgcaaagg gacttttcag
     9301 gctagccgcc agccgggtaa tcagcttatc cagcaacgtt tcgctggatg ttggcggcaa
     9361 cgaatcactg gtgtaacgat ggcgattcag caacatcacc aactgcccga acagcaactc
     9421 agccatttcg ttagcaaacg gcacatgctg actactttca tgctcaagct gaccgataac
     9481 ctgccgcgcc tgcgccatcc ccatgctacc taagcgccag tgtggttgcc ctgcgctggc
     9541 gttaaatccc ggaatcgccc cctgccagtc aagattcagc ttcagacgct ccgggcaata
     9601 aataatattc tgcaaaacca gatcgttaac ggaagcgtag gagtgtttat cgtcagcatg
     9661 aatgtaaaag agatcgccac gggtaatgcg ataagggcga tcgttgagta catgcaggcc
     9721 attaccgcgc cagacaatca ccagctcaca aaaatcatgt gtatgttcag caaagacatc
     9781 ttgcggataa cggtcagcca cagcgactgc ctgctggtcg ctggcaaaaa aatcatcttt
     9841 gagaagtttt aactgatgcg ccaccgtggc tacctcggcc agagaacgaa gttgattatt
     9901 cgcaatatgg cgtacaaata cgttgagaag attcgcgtta ttgcagaaag ccatcccgtc
     9961 cctggcgaat atcacgcggt gaccagttaa actctcggcg aaaaagcgtc gaaaagtggt
    10021 tactgtcgct gaatccacag cgataggcga tgtcagtaac gctggcctcg ctgtggcgta
    10081 gcagatgtcg ggctttcatc agtcgcaggc ggttcaggta tcgctgaggc gtcagtcccg
    10141 tttgctgctt aagctgccga tgtagcgtac gcagtgaaag agaaaattga tccgccacgg
    10201 catcccaatt cacctcatcg gcaaaatggt cctccagcca ggccagaagc aagttgagac
    10261 gtgatgcgct gttttccagg ttctcctgca aactgctttt acgcagcaag agcagtaatt
    10321 gcataaacaa gatctcgcga ctggcggtcg agggtaaatc attttcccct tcctgctgtt
    10381 ccatctgtgc aaccagctgt cgcacctgct gcaatacgct gtggttaacg cgccagtgag
    10441 acggatactg cccatccagc tcttgtggca gcaactgatt cagcccggcg agaaactgaa
    10501 atcgatccgg cgagcgatac agcacattgg tcagacacag attatcggta tgttcataca
    10561 gatgccgatc atgatcgcgt acgaaacaga ccgtgccgcc ggtgatggta tagggctgcc
    10621 cattaaacac atgaataccc gtgccatgtt cgacaatcac aatttcatga aaatcatgat
    10681 gatgttcagg aaaatccgcc tgcgggagcc ggggttctat cgccacggac gcgttaccag
    10741 acggaaaaaa atccacacta tgtaatacgg tcatactggc ctcctgatgt cgtcaa
//
