LOCUS       sequence-221057-       10256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A cytidine deaminase-mediated base-editing plasmid in Klebsiella 
            Pneumoniae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10256)
  AUTHORS   Wang Y, Wang S, Chen W, Song L, Zhang Y, Shen Z, Yu F, Li M, Ji Q
  TITLE     Precise and efficient genome editing in Klebsiella pneumoniae using 
            CRISPR-Cas9 and CRISPR-assisted cytidine deaminase.
  JOURNAL   Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 
            10.1128/AEM.01834-18.
  PUBMED    30217854
REFERENCE   2  (bases 1 to 10256)
  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: "Appl 
            Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 
            10.1128/AEM.01834-18."
COMMENT     Sequence Label: pBECKP-km
FEATURES             Location/Qualifiers
     source          1..10256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..18
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2..18
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     55..71
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     56..72
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(134..1468,1469..1555))
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             2100..2786
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(2835..2858,2859..2861,2862..6938)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     promoter        6983..7017
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        7035..7110
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(7143..7159)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7143..7159)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7218..7234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7218..7234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             8018..8209
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(8203..8225)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    8311..8453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8468..8485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8639..9227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8719..8738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(9338..9357)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             9349..10164
                     /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"
     CDS             9349..10164
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /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     complement(9422..9441)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tgtaaaacga cggccagtga attgacgcgt attgggatgg taccgggccc cccctcgagg
       61 tcgacggtat cgatacggca tcagagcaga ttgtactgag agtgcaccat aatcggcatt
      121 ttcttttgcg tttttatttg ttaactgtta attgtccttg ttcaaggatg ctgtctttga
      181 caacagatgt tttcttgcct ttgatgttca gcaggaagct aggcgcaaac gttgattgtt
      241 tgtctgcgta gaatcctctg tttgtcatat agcttgtaat cacgacattg tttcctttcg
      301 cttgaggtac agcgaagtgt gagtaagtaa aggttacatc gttaggatca agatccattt
      361 ttaacacaag gccagttttg ttcagcggct tgtatgggcc agttaaagaa ttagaaacat
      421 aaccaagcat gtaaatatcg ttagacgtaa tgccgtcaat cgtcattttt gatccgcggg
      481 agtcagtgaa cagataccat ttgccgttca ttttaaagac gttcgcgcgt tcaatttcat
      541 ctgttactgt gttagatgca atcagcggtt tcatcacttt tttcagtgtg taatcatcgt
      601 ttagctcaat cataccgaga gcgccgtttg ctaactcagc cgtgcgtttt ttatcgcttt
      661 gcagaagttt ttgactttct tgacggaaga atgatgtgct tttgccatag tatgctttgt
      721 taaataaaga ttcttcgcct tggtagccat cttcagttcc agtgtttgct tcaaatacta
      781 agtatttgtg gcctttatct tctacgtagt gaggatctct cagcgtatgg ttgtcgcctg
      841 agctgtagtt gccttcatcg atgaactgct gtacattttg atacgttttt ccgtcaccgt
      901 caaagattga tttataatcc tctacaccgt tgatgttcaa agagctgtct gatgctgata
      961 cgttaacttg tgcagttgtc agtgtttgtt tgccgtaatg tttaccggag aaatcagtgt
     1021 agaataaacg gatttttccg tcagatgtaa atgtggctga acctgaccat tcttgtgttt
     1081 ggtcttttag gatagaatca tttgcatcga atttgtcgct gtctttaaag acgcggccag
     1141 cgtttttcca gctgtcaata gaagtttcgc cgactttttg atagaacatg taaatcgatg
     1201 tgtcatccgc atttttagga tctccggcta atgcaaagac gatgtggtag ccgtgatagt
     1261 ttgcgacagt gccgtcagcg ttttgtaatg gccagctgtc ccaaacgtcc aggccttttg
     1321 cagaagagat atttttaatt gtggacgaat cgaactcagg aacttgatat ttttcatttt
     1381 tttgctgttc agggatttgc agcatatcat ggcgtgtaat atgggaaatg ccgtatgttt
     1441 ccttatatgg cttttggttc gtttctttcg caaacgcttg agttgcgcct cctgccagca
     1501 gtgcggtagt aaaggttaat actgttgctt gttttgcaaa ctttttgatg ttcatcgttc
     1561 atgtctcctt ttttatgtac tgtgttagcg gtctgcttct tccagccctc ctgtttgaag
     1621 atggcaagtt agttacgcac aataaaaaaa gacctaaaat atgtaagggg tgacgccaaa
     1681 gtatacactt tgccctttac acattttagg tcttgcctgc tttatcagta acaaacccgc
     1741 gcgatttact tttcgacctc attctattag actctcgttt ggattgcaac tggtctattt
     1801 tcctcttttg tttgatagaa aatcataaaa ggatttgcag actacgggcc taaagaacta
     1861 aaaaatctat ctgtttcttt tcattctctg tattttttat agtttctgtt gcatgggcat
     1921 aaagttgcaa gcttgcaaac atgctttggg taaacatgta ggtattaacg tcaatgcgac
     1981 aatagtagca ttgattaaat gagaattagt aagtgtttta cttactaaat tttatttaac
     2041 ctaaaaatga accacctgga tgtgtgggat taaaaagtga agagaggagg acatatcaca
     2101 tgagtagcga aaccggtccg gttgcagtgg atccgaccct gcgccgtcgc attgaaccgc
     2161 acgagtttga agtgttcttt gatccgcgcg agctgcgcaa agaaacttgc ctgctgtacg
     2221 agattaactg gggtggccgc catagcatct ggcgccatac cagccagaac accaataagc
     2281 acgtggaagt gaattttatt gaaaaattta ccaccgagcg ctacttctgc cctaataccc
     2341 gctgcagcat cacctggttt ctgagctgga gcccgtgcgg cgaatgtagt cgcgccatta
     2401 ccgagttcct gagccgctat ccgcatgtga ccctgttcat ctacatcgcc cgtctgtacc
     2461 atcacgccga tccgcgcaat cgccaaggtc tgcgtgatct gattagcagc ggtgtgacca
     2521 tccagatcat gaccgaacaa gagagcggct attgctggcg caacttcgtg aactattctc
     2581 cgagcaacga agcccactgg ccgcgttatc cgcatctgtg ggtgcgcctg tatgtgctgg
     2641 agctgtactg catcatcctg ggcctgccgc cttgcctgaa tattctgcgc cgtaaacagc
     2701 cgcaactgac attcttcacc atcgcactgc agagctgcca ttatcagcgc ctgccgccgc
     2761 acattttatg ggccaccggt ctgaaaagcg gtagtgaaac tccgggcaca agcgaaagcg
     2821 caaccccgga aagtgataag aaatactcaa taggcttagc tatcggcaca aatagcgtcg
     2881 gatgggcggt gatcactgat gaatataagg ttccgtctaa aaagttcaag gttctgggaa
     2941 atacagaccg ccacagtatc aaaaaaaatc ttataggggc tcttttattt gacagtggag
     3001 agacagcgga agcgactcgt ctcaaacgga cagctcgtag aaggtataca cgtcggaaga
     3061 atcgtatttg ttatctacag gagatttttt caaatgagat ggcgaaagta gatgatagtt
     3121 tctttcatcg acttgaagag tcttttttgg tggaagaaga caagaagcat gaacgtcatc
     3181 ctatttttgg aaatatagta gatgaagttg cttatcatga gaaatatcca actatctatc
     3241 atctgcgaaa aaaattggta gattctactg ataaagcgga tttgcgctta atctatttgg
     3301 ccttagcgca tatgattaag tttcgtggtc attttttgat tgagggagat ttaaatcctg
     3361 ataatagtga tgtggacaaa ctatttatcc agttggtaca aacctacaat caattatttg
     3421 aagaaaaccc tattaacgca agtggagtag atgctaaagc gattctttct gcacgattga
     3481 gtaaatcaag acgattagaa aatctcattg ctcagctccc cggtgagaag aaaaatggct
     3541 tatttgggaa tctcattgct ttgtcattgg gtttgacccc taattttaaa tcaaattttg
     3601 atttggcaga agatgctaaa ttacagcttt caaaagatac ttacgatgat gatttagata
     3661 atttattggc gcaaattgga gatcaatatg ctgatttgtt tttggcagct aagaatttat
     3721 cagatgctat tttactttca gatatcctaa gagtaaatac tgaaataact aaggctcccc
     3781 tatcagcttc aatgattaaa cgctacgatg aacatcatca agacttgact cttttaaaag
     3841 ctttagttcg acaacaactt ccagaaaagt ataaagaaat cttttttgat caatcaaaaa
     3901 acggatatgc aggttatatt gatgggggag ctagccaaga agaattttat aaatttatca
     3961 aaccaatttt agaaaaaatg gatggtactg aggaattatt ggtgaaacta aatcgtgaag
     4021 atttgctgcg caagcaacgg acctttgaca acggctctat tccccatcaa attcacttgg
     4081 gtgagctgca tgctattttg agaagacaag aagactttta tccattttta aaagacaatc
     4141 gtgagaagat tgaaaaaatc ttgacttttc gaattcctta ttatgttggt ccattggcgc
     4201 gtggcaatag tcgttttgca tggatgactc ggaagtctga agaaacaatt accccatgga
     4261 attttgaaga agttgtcgat aaaggtgctt cagctcaatc atttattgaa cgcatgacaa
     4321 actttgataa aaatcttcca aatgaaaaag tactaccaaa acatagtttg ctttatgagt
     4381 attttacggt ttataacgaa ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac
     4441 cagcatttct ttcaggtgaa cagaagaaag ccattgttga tttactcttc aaaacaaatc
     4501 gaaaagtaac cgttaagcaa ttaaaagaag attatttcaa aaaaatagaa tgttttgata
     4561 gtgttgaaat ttcaggagtt gaagatagat ttaatgcttc attaggtacc taccatgatt
     4621 tgctaaaaat tattaaagat aaagattttt tggataatga agaaaatgaa gatatcttag
     4681 aggatattgt tttaacattg accttatttg aagataggga gatgattgag gaaagactta
     4741 aaacatatgc tcacctcttt gatgataagg tgatgaaaca gcttaaacgt cgccgttata
     4801 ctggttgggg acgtttgtct cgaaaattga ttaatggtat tagggataag caatctggca
     4861 aaacaatatt agattttttg aaatcagatg gttttgccaa tcgcaatttt atgcagctga
     4921 tccatgatga tagtttgaca tttaaagaag acattcaaaa agcacaagtg tctggacaag
     4981 gcgatagttt acatgaacat attgcaaatt tagctggtag ccctgctatt aaaaaaggta
     5041 ttttacagac tgtaaaagtt gttgatgaat tggtcaaagt aatggggcgg cataagccag
     5101 aaaatatcgt tattgaaatg gcacgtgaaa atcagacaac tcaaaagggc cagaaaaatt
     5161 cgcgagagcg tatgaaacga atcgaagaag gtatcaaaga attaggaagt cagattctta
     5221 aagagcatcc tgttgaaaat actcaattgc aaaatgaaaa gctctatctc tattatctcc
     5281 aaaatggaag agacatgtat gtggaccaag aattagatat taatcgttta agtgattatg
     5341 atgtcgatca cattgttcca caaagtttcc ttaaagacga ttcaatagac aataaggtct
     5401 taacgcgttc tgataaaaat cgtggtaaat cggataacgt tccaagtgaa gaagtagtca
     5461 aaaagatgaa aaactattgg agacaacttc taaacgccaa gttaatcact caacgtaagt
     5521 ttgataattt aacgaaagct gaacgtggag gtttgagtga acttgataaa gctggtttta
     5581 tcaaacgcca attggttgaa actcgccaaa tcactaagca tgtggcacaa attttggata
     5641 gtcgcatgaa tactaaatac gatgaaaatg ataaacttat tcgagaggtt aaagtgatta
     5701 ccttaaaatc taaattagtt tctgacttcc gaaaagattt ccaattctat aaagtacgtg
     5761 agattaacaa ttaccatcat gcccatgatg cgtatctaaa tgccgtcgtt ggaactgctt
     5821 tgattaagaa atatccaaaa cttgaatcgg agtttgtcta tggtgattat aaagtttatg
     5881 atgttcgtaa aatgattgct aagtctgagc aagaaatagg caaagcaacc gcaaaatatt
     5941 tcttttactc taatatcatg aacttcttca aaacagaaat tacacttgca aatggagaga
     6001 ttcgcaaacg ccctctaatc gaaactaatg gggaaactgg agaaattgtc tgggataaag
     6061 ggcgagattt tgccacagtg cgcaaagtat tgtccatgcc ccaagtcaat attgtcaaga
     6121 aaacagaagt acagacaggc ggattctcca aggagtcaat tttaccaaaa agaaattcgg
     6181 acaagcttat tgctcgtaaa aaagactggg atccaaaaaa atatggtggt tttgatagtc
     6241 caacggtagc ttattcagtc ctagtggttg ctaaggtgga aaaagggaaa tcgaagaagt
     6301 taaaatccgt taaagagtta ctagggatca caattatgga aagaagttcc tttgaaaaaa
     6361 atccgattga ctttttagaa gctaaaggat ataaggaagt taaaaaagac ttaatcatta
     6421 aactacctaa atatagtctt tttgagttag aaaacggtcg taaacggatg ctggctagtg
     6481 ccggagaatt acaaaaagga aatgagctgg ctctgccaag caaatatgtg aattttttat
     6541 atttagctag tcattatgaa aagttgaagg gtagtccaga agataacgaa caaaaacaat
     6601 tgtttgtgga gcagcataag cattatttag atgagattat tgagcaaatc agtgaatttt
     6661 ctaagcgtgt tattttagca gatgccaatt tagataaagt tcttagtgca tataacaaac
     6721 atagagacaa accaatacgt gaacaagcag aaaatattat tcatttattt acgttgacga
     6781 atcttggagc tcccgctgct tttaaatatt ttgatacaac aattgatcgt aaacgatata
     6841 cgtctacaaa agaagtttta gatgccactc ttatccatca atccatcact ggtctttatg
     6901 aaacacgcat tgatttgagt cagctaggag gtgactgatg gctggttggc gtactgttgt
     6961 ggtaaatacc aagcttgata tcttgacagc tagctcagtc ctaggtataa tactagtcga
     7021 gaccattggt ctcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     7081 caacttgaaa aagtggcacc gagtcggtgc tttttttgat atcgaattcc tgcagcccgg
     7141 gggatccact agttctagag cggccgccac cgcggtggag ctcatcccaa tggcgcgccg
     7201 agcttggctc gagcatggtc atagctgttt cctgctcact cattaggcac cgggatctcg
     7261 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     7321 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     7381 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     7441 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     7501 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg
     7561 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     7621 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     7681 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     7741 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
     7801 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
     7861 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
     7921 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     7981 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     8041 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     8101 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     8161 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     8221 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     8281 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     8341 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     8401 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     8461 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     8521 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     8581 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     8641 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     8701 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     8761 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8821 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8881 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8941 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     9001 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     9061 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     9121 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     9181 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     9241 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     9301 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     9361 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
     9421 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     9481 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     9541 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     9601 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa
     9661 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     9721 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     9781 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     9841 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     9901 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     9961 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
    10021 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
    10081 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
    10141 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
    10201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
