pBECKP-km

LOCUS       pBECKP-km           10252 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  A cytidine deaminase-mediated base-editing plasmid in Klebsiella 
            Pneumoniae.
ACCESSION   .
VERSION     .
KEYWORDS    pBECKP-km
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10252)
  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 10252)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10252
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..687
                     /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             736..4839
                     /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        4884..4918
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        4936..5011
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(5044..5060)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5044..5060)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5119..5135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5119..5135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5919..6110
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6104..6126)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6212..6351
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6366..6383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6537..7125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6617..6636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7236..7255)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7247..8062
                     /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(7320..7339)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8208..8224
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8209..8225
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(8287..9708)
                     /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"
ORIGIN
        1 atgagtagcg aaaccggtcc ggttgcagtg gatccgaccc tgcgccgtcg cattgaaccg
       61 cacgagtttg aagtgttctt tgatccgcgc gagctgcgca aagaaacttg cctgctgtac
      121 gagattaact ggggtggccg ccatagcatc tggcgccata ccagccagaa caccaataag
      181 cacgtggaag tgaattttat tgaaaaattt accaccgagc gctacttctg ccctaatacc
      241 cgctgcagca tcacctggtt tctgagctgg agcccgtgcg gcgaatgtag tcgcgccatt
      301 accgagttcc tgagccgcta tccgcatgtg accctgttca tctacatcgc ccgtctgtac
      361 catcacgccg atccgcgcaa tcgccaaggt ctgcgtgatc tgattagcag cggtgtgacc
      421 atccagatca tgaccgaaca agagagcggc tattgctggc gcaacttcgt gaactattct
      481 ccgagcaacg aagcccactg gccgcgttat ccgcatctgt gggtgcgcct gtatgtgctg
      541 gagctgtact gcatcatcct gggcctgccg ccttgcctga atattctgcg ccgtaaacag
      601 ccgcaactga cattcttcac catcgcactg cagagctgcc attatcagcg cctgccgccg
      661 cacattttat gggccaccgg tctgaaaagc ggtagtgaaa ctccgggcac aagcgaaagc
      721 gcaaccccgg aaagtgataa gaaatactca ataggcttag ctatcggcac aaatagcgtc
      781 ggatgggcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa ggttctggga
      841 aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt tgacagtgga
      901 gagacagcgg aagcgactcg tctcaaacgg acagctcgta gaaggtatac acgtcggaag
      961 aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt agatgatagt
     1021 ttctttcatc gacttgaaga gtcttttttg gtggaagaag acaagaagca tgaacgtcat
     1081 cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc aactatctat
     1141 catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt aatctatttg
     1201 gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga tttaaatcct
     1261 gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa tcaattattt
     1321 gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc tgcacgattg
     1381 agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc
     1441 ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa atcaaatttt
     1501 gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga tgatttagat
     1561 aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc taagaattta
     1621 tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac taaggctccc
     1681 ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac tcttttaaaa
     1741 gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga tcaatcaaaa
     1801 aacggatatg caggttatat tgatggggga gctagccaag aagaatttta taaatttatc
     1861 aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact aaatcgtgaa
     1921 gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca aattcacttg
     1981 ggtgagctgc atgctatttt gagaagacaa gaagactttt atccattttt aaaagacaat
     2041 cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt attatgttgg tccattggcg
     2101 cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat taccccatgg
     2161 aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga acgcatgaca
     2221 aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt gctttatgag
     2281 tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa
     2341 ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt caaaacaaat
     2401 cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga atgttttgat
     2461 agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac ctaccatgat
     2521 ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga agatatctta
     2581 gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga ggaaagactt
     2641 aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg tcgccgttat
     2701 actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa gcaatctggc
     2761 aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt tatgcagctg
     2821 atccatgatg atagtttgac atttaaagaa gacattcaaa aagcacaagt gtctggacaa
     2881 ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat taaaaaaggt
     2941 attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg gcataagcca
     3001 gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat
     3061 tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag tcagattctt
     3121 aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct ctattatctc
     3181 caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt aagtgattat
     3241 gatgtcgatc acattgttcc acaaagtttc cttaaagacg attcaataga caataaggtc
     3301 ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga agaagtagtc
     3361 aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac tcaacgtaag
     3421 tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa agctggtttt
     3481 atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca aattttggat
     3541 agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt taaagtgatt
     3601 accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta taaagtacgt
     3661 gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt tggaactgct
     3721 ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta taaagtttat
     3781 gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac cgcaaaatat
     3841 ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc aaatggagag
     3901 attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt ctgggataaa
     3961 gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag
     4021 aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa aagaaattcg
     4081 gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg ttttgatagt
     4141 ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag
     4201 ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc ctttgaaaaa
     4261 aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga cttaatcatt
     4321 aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat gctggctagt
     4381 gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt gaatttttta
     4441 tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga acaaaaacaa
     4501 ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat cagtgaattt
     4561 tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc atataacaaa
     4621 catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt tacgttgacg
     4681 aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg taaacgatat
     4741 acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac tggtctttat
     4801 gaaacacgca ttgatttgag tcagctagga ggtgactgat ggctggttgg cgtactgttg
     4861 tggtaaatac caagcttgat atcttgacag ctagctcagt cctaggtata atactagtcg
     4921 agaccattgg tctcagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     4981 tcaacttgaa aaagtggcac cgagtcggtg ctttttttga tatcgaattc ctgcagcccg
     5041 ggggatccac tagttctaga gcggccgcca ccgcggtgga gctcatccca atggcgcgcc
     5101 gagcttggct cgagcatggt catagctgtt tcctgctcac tcattaggca ccgggatctc
     5161 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     5221 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     5281 cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc
     5341 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg
     5401 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc ccacgggtgc
     5461 gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt
     5521 agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg
     5581 cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc
     5641 ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac
     5701 cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc
     5761 tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg
     5821 catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta
     5881 cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg
     5941 cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg
     6001 agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc
     6061 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6121 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6181 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     6241 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6301 ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     6361 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6421 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6481 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6541 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6601 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6661 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6721 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6781 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6841 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6901 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6961 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7021 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7081 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7141 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7201 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     7261 acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg
     7321 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     7381 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     7441 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     7501 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac
     7561 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     7621 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     7681 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     7741 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     7801 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     7861 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     7921 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     7981 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     8041 gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt
     8101 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtgtaaa
     8161 cgacggccag tgaattgacg cgtattggga tggtaccggg ccccccctcg aggtcgacgg
     8221 tatcgatacg gcatcagagc agattgtact gagagtgcac cataatcggc attttctttt
     8281 gcgtttttat ttgttaactg ttaattgtcc ttgttcaagg atgctgtctt tgacaacaga
     8341 tgttttcttg cctttgatgt tcagcaggaa gctaggcgca aacgttgatt gtttgtctgc
     8401 gtagaatcct ctgtttgtca tatagcttgt aatcacgaca ttgtttcctt tcgcttgagg
     8461 tacagcgaag tgtgagtaag taaaggttac atcgttagga tcaagatcca tttttaacac
     8521 aaggccagtt ttgttcagcg gcttgtatgg gccagttaaa gaattagaaa cataaccaag
     8581 catgtaaata tcgttagacg taatgccgtc aatcgtcatt tttgatccgc gggagtcagt
     8641 gaacagatac catttgccgt tcattttaaa gacgttcgcg cgttcaattt catctgttac
     8701 tgtgttagat gcaatcagcg gtttcatcac ttttttcagt gtgtaatcat cgtttagctc
     8761 aatcataccg agagcgccgt ttgctaactc agccgtgcgt tttttatcgc tttgcagaag
     8821 tttttgactt tcttgacgga agaatgatgt gcttttgcca tagtatgctt tgttaaataa
     8881 agattcttcg ccttggtagc catcttcagt tccagtgttt gcttcaaata ctaagtattt
     8941 gtggccttta tcttctacgt agtgaggatc tctcagcgta tggttgtcgc ctgagctgta
     9001 gttgccttca tcgatgaact gctgtacatt ttgatacgtt tttccgtcac cgtcaaagat
     9061 tgatttataa tcctctacac cgttgatgtt caaagagctg tctgatgctg atacgttaac
     9121 ttgtgcagtt gtcagtgttt gtttgccgta atgtttaccg gagaaatcag tgtagaataa
     9181 acggattttt ccgtcagatg taaatgtggc tgaacctgac cattcttgtg tttggtcttt
     9241 taggatagaa tcatttgcat cgaatttgtc gctgtcttta aagacgcggc cagcgttttt
     9301 ccagctgtca atagaagttt cgccgacttt ttgatagaac atgtaaatcg atgtgtcatc
     9361 cgcattttta ggatctccgg ctaatgcaaa gacgatgtgg tagccgtgat agtttgcgac
     9421 agtgccgtca gcgttttgta atggccagct gtcccaaacg tccaggcctt ttgcagaaga
     9481 gatattttta attgtggacg aatcgaactc aggaacttga tatttttcat ttttttgctg
     9541 ttcagggatt tgcagcatat catggcgtgt aatatgggaa atgccgtatg tttccttata
     9601 tggcttttgg ttcgtttctt tcgcaaacgc ttgagttgcg cctcctgcca gcagtgcggt
     9661 agtaaaggtt aatactgttg cttgttttgc aaactttttg atgttcatcg ttcatgtctc
     9721 cttttttatg tactgtgtta gcggtctgct tcttccagcc ctcctgtttg aagatggcaa
     9781 gttagttacg cacaataaaa aaagacctaa aatatgtaag gggtgacgcc aaagtataca
     9841 ctttgccctt tacacatttt aggtcttgcc tgctttatca gtaacaaacc cgcgcgattt
     9901 acttttcgac ctcattctat tagactctcg tttggattgc aactggtcta ttttcctctt
     9961 ttgtttgata gaaaatcata aaaggatttg cagactacgg gcctaaagaa ctaaaaaatc
    10021 tatctgtttc ttttcattct ctgtattttt tatagtttct gttgcatggg cataaagttg
    10081 caagcttgca aacatgcttt gggtaaacat gtaggtatta acgtcaatgc gacaatagta
    10141 gcattgatta aatgagaatt agtaagtgtt ttacttacta aattttattt aacctaaaaa
    10201 tgaaccacct ggatgtgtgg gattaaaaag tgaagagagg aggacatatc ac
//