LOCUS       sequence-148602-       11705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genome editing for gene xylR (cbei-2385) in clostridium beijerinckii
            NCIMB 8052.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11705)
  AUTHORS   Li Q, Chen J, Minton NP, Zhang Y, Wen Z, Liu J, Yang H, Zeng Z, Ren 
            X, Yang J, Gu Y, Jiang W, Jiang Y, Yang S
  TITLE     CRISPR-based genome editing and expression control systems in 
            Clostridium acetobutylicum and Clostridium beijerinckii.
  JOURNAL   Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053.
  PUBMED    27213844
REFERENCE   2  (bases 1 to 11705)
  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: "Biotechnol 
            J. 2016 May 23. doi: 10.1002/biot.201600053."
COMMENT     Sequence Label: pNICKclos2.0
FEATURES             Location/Qualifiers
     source          1..11705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1487..1521
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1542..1617
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(3803..3829,3830..3832,3833..7909)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(7926..7943)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7926..7942)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7935..7957)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(8151..8170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8270..8292
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8330..8348)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8416..8520
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8521..8589,8590..9381)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8739..8758)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9552..10140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10041..10060
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10294..10311
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10428..10449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10464..10469,10470..10487,10488..10494)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10502..10518
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10507..10529
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10526..10542
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10526..10542
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgataattta cagaaaagaa aattatagaa tttagtatga ttaattatac tcatttatga
       61 atgtttaatt gaatacaaaa aaaaatactt gttatgtatt caattacggg ttaaaatata
      121 gacaagttga aaaatttaat aaaaaaataa gtcctcagct cttatatatt aagctaccaa
      181 cttagtatat aagccaaaac ttaaatgtgc taccaacaca tcaagccgtt agagaactct
      241 atctatagca atatttcaaa tgtaccgaca tacaagagaa acattaacta tatatattca
      301 atttatgaga ttatcttaac agatataaat gtaaattgca ataagtaaga tttagaagtt
      361 tatagccttt gtgtattgga agcagtacgc aaaggctttt ttatttgata aaaattagaa
      421 gtatatttat tttttcataa ttaatttatg aaaatgaaag ggggtgagca aagtgacaga
      481 ggaaagcagt atcttatcaa ataacaaggt attagcaata tcattattga ctttagcagt
      541 aaacattatg acttttatag tgcttgtagc taagtagtac gaaaggggga gctttaaaaa
      601 gctccttgga atacatagaa ttcataaatt aatttatgaa aagaagggcg tatatgaaaa
      661 cttgtaaaaa ttgcaaagag tttattaaag atactgaaat atgcaaaata cattcgttga
      721 tgattcatga taaaacagta gcaacctatt gcagtaaata caatgagtca agatgtttac
      781 ataaagggaa agtccaatgt attaattgtt caaagatgaa ccgatatgga tggtgtgcca
      841 taaaaatgag atgttttaca gaggaagaac agaaaaaaga acgtacatgc attaaatatt
      901 atgcaaggag ctttaaaaaa gctcatgtaa agaagagtaa aaagaaaaaa taatttattt
      961 attaatttaa tattgagagt gccgacacag tatgcactaa aaaatatatc tgtggtgtag
     1021 tgagccgata caaaaggata gtcactcgca ttttcataat acatcttatg ttatgattat
     1081 gtgtcggtgg gacttcacga cgaaaaccca caataaaaaa agagttcggg gtagggttaa
     1141 gcatagttga ggcaactaaa caatcaagct aggatatgca gtagcagacc gtaaggtcgt
     1201 tgtttaggtg tgttgtaata catacgctat taagatgtaa aaatacggat accaatgaag
     1261 ggaaaagtat aatttttgga tgtagtttgt ttgttcatct atgggcaaac tacgtccaaa
     1321 gccgtttcca aatctgctaa aaagtatatc ctttctaaaa tcaaagtcaa gtatgaaatc
     1381 ataaataaag tttaattttg aagttattat gatattatgt ttttctatta aaataaatta
     1441 agtatataga atagtttaat aatagtatat acttaatgtg ctgcagttga cagctagctc
     1501 agtcctaggt ataatactag tcgagttaga cataatagtg agttttagag ctagaaatag
     1561 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     1621 tttgcggccg caaaacctgg ttctaatgga attgtttact taccatatct tatgggagaa
     1681 agaacacctc atatagaccc taatgtaaaa ggtgcatttt taggagtatc actaataaat
     1741 aaccatgatg attttgtacg tagtatatta gaaggtgtta gctttagttt gaaaaactgc
     1801 cttgatttaa tcgaagacat gaatgtaaac attgatgaaa taagagtaag tggcggtgga
     1861 gcagaaagtg ctgtatggag acaaatatta gctgatattt tccaatattc attaactaca
     1921 gttaaagcat ctgaaggcgg tgcgcttgga gttgcaatat tagcaggggt tggagcagga
     1981 atctatgatt ctgtagaaga tgcttgtgat agaatcgtaa aaggaaacaa gaaagtttct
     2041 ccaaatgaag aattaaaaga agtatactca aaagtatatg aaacatataa ttcagcttat
     2101 ccaagaataa aagatattta aatatattaa agatagaaat caatcatctt agttaagtaa
     2161 ttgtataatt aattaagggt gtgtagtagt atcgaatttt agcaaaaaaa tcatatagtt
     2221 ttttaaatac tttaacaaaa ataataatag tgatattatt taagaaaaag ctactattac
     2281 tgataatagt cgagaggagt attttaaagt ttgatagaag taaatcacag taaaataaaa
     2341 gaaactaaca gaaaaaagat aatagcatta ttattaaaga aaaatgagat caccaaatta
     2401 gatatatcta gaatattaga tataagtata acaacagttt ctacaaatat tacagaattg
     2461 aaaagcgaag gtattgtaga agatgtaaga tcgttggaat ctacaggagg aagaaaagca
     2521 attgctatta agttgaatga aaactgtaga tattcaattg gagtcgcact aacaccgaat
     2581 catataaaga tatcattagt aaatataaag aaaaaagtta ttgaaagtat gctgcagtgg
     2641 aatagagaac ataataaatt tacttaatga aaatatagat ttgttaatga agaaatataa
     2701 tttatgctct gcaaatttat taggaatagg gatttcatta cctggaaccg tagattttaa
     2761 agaaggcata ataaagtatt catatttgtt aggcgccaaa gattttaatt taaaggaaaa
     2821 gttcgaatac ttagacattc cagtatatgt agataatgaa gcaaatttat ctgcttacta
     2881 cgagttttta aataaaagag atatactaag aaatctatta tatgtatcaa taacagaagg
     2941 tttaggcctt ggtataataa taaatggaaa aatttacaga ggtgacaata actcatcagg
     3001 agaattaggt catactaaga tagcaattga tggaaaaaaa tgtaagtgtg gagctagagg
     3061 ctgtttagaa gcatacacat ctatgaattc tctgattgat agttataatg aagctaatag
     3121 ttcaaatatt agtgatattg atgaatttga agaaagatat aatcaaaatg acaaagatgc
     3181 tcatgatgta ttaaatgatt atttaaaaac attgtcttta ggtctatcga atttggttat
     3241 gatatttgat cctagcagca tagtaatagg tggagatata aataatttac ttaatgataa
     3301 gatagatatg ctgaaaaaag aggtttataa agataattta tttacaaatg aaaataattg
     3361 cagtataagt attgcaagtt ttaaagaatc ttatctacta ggagctgcta tgatgcctat
     3421 agaagaattc ttagaaatta aataatggag tggtgaaagc tgctctatta atttaaaaac
     3481 ttattaaata aatttttaaa atatattttt aaagttatgt aagcatagct atattaaggg
     3541 gggtcttaat ctttgattgg aactccagat taagacaagc cttttatata aaactaaata
     3601 tctatttatt tcatattaga tttatgttct tgattttctc gagtttttaa caaaatatat
     3661 tgataaaaat aatattagtg ggtataatta agttgttaga gaaaacgtat aaattaggga
     3721 taaactatgg aacttatgaa atagattgaa atggtttatc tgttaccccg tatcaaaatt
     3781 taggaggtta gttagacccg ggatggataa gaaatactca ataggcttag ctatcggcac
     3841 aaatagcgtc ggatgggcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa
     3901 ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt
     3961 tgacagtgga gagacagcgg aagcgactcg tctcaaacgg acagctcgta gaaggtatac
     4021 acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt
     4081 agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag acaagaagca
     4141 tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc
     4201 aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt
     4261 aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga
     4321 tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa
     4381 tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc
     4441 tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa
     4501 gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa
     4561 atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga
     4621 tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc
     4681 taagaattta tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac
     4741 taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac
     4801 tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga
     4861 tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag aagaatttta
     4921 taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact
     4981 aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca
     5041 aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt atccattttt
     5101 aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt attatgttgg
     5161 tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat
     5221 taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga
     5281 acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt
     5341 gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg
     5401 aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt
     5461 caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga
     5521 atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac
     5581 ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga
     5641 agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga
     5701 ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg
     5761 tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa
     5821 gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt
     5881 tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa aagcacaagt
     5941 gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat
     6001 taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg
     6061 gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg
     6121 ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag
     6181 tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct
     6241 ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt
     6301 aagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg attcaataga
     6361 caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga
     6421 agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac
     6481 tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa
     6541 agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca
     6601 aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt
     6661 taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta
     6721 taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt
     6781 tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta
     6841 taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac
     6901 cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc
     6961 aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt
     7021 ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa
     7081 tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa
     7141 aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg
     7201 ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa
     7261 atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc
     7321 ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga
     7381 cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat
     7441 gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt
     7501 gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga
     7561 acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat
     7621 cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc
     7681 atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt
     7741 tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg
     7801 taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac
     7861 tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgactgag cggccgcggg
     7921 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     7981 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     8041 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     8101 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     8161 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     8221 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     8281 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     8341 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     8401 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     8461 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     8521 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     8581 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     8641 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     8701 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     8761 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     8821 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     8881 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8941 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     9001 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     9061 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     9121 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     9181 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     9241 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     9301 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     9361 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     9421 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     9481 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     9541 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     9601 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     9661 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     9721 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     9781 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     9841 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9901 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9961 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
    10021 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
    10081 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
    10141 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
    10201 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
    10261 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
    10321 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
    10381 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
    10441 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
    10501 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
    10561 tggctaacac acacgccatt ccaaccaata gttttctcgg cataaagcca tgctctgacg
    10621 cttaaatgca ctaatgcctt aaaaaaacat taaagtctaa cacactagac ttatttactt
    10681 cgtaattaag tcgttaaacc gtgtgctcta cgaccaaaag tataaaacct ttaagaactt
    10741 tcttttttct tgtaaaaaaa gaaactagat aaatctctca tatcttttat tcaataatcg
    10801 catcagattg cagtataaat ttaacgatca ctcatcatgt tcatatttat cagagctcgt
    10861 gctataatta tactaatttt ataaggagga aaaaataaag agggttataa tgaacgagaa
    10921 aaatataaaa cacagtcaaa actttattac ttcaaaacat aatatagata aaataatgac
    10981 aaatataaga ttaaatgaac atgataatat ctttgaaatc ggctcaggaa aagggcattt
    11041 tacccttgaa ttagtacaga ggtgtaattt cgtaactgcc attgaaatag accataaatt
    11101 atgcaaaact acagaaaata aacttgttga tcacgataat ttccaagttt taaacaagga
    11161 tatattgcag tttaaatttc ctaaaaacca atcctataaa atatttggta atatacctta
    11221 taacataagt acggatataa tacgcaaaat tgtttttgat agtatagctg atgagattta
    11281 tttaatcgtg gaatacgggt ttgctaaaag attattaaat acaaaacgct cattggcatt
    11341 atttttaatg gcagaagttg atatttctat attaagtatg gttccaagag aatattttca
    11401 tcctaaacct aaagtgaata gctcacttat cagattaaat agaaaaaaat caagaatatc
    11461 acacaaagat aaacagaagt ataattattt cgttatgaaa tgggttaaca aagaatacaa
    11521 gaaaatattt acaaaaaatc aatttaacaa ttccttaaaa catgcaggaa ttgacgattt
    11581 aaacaatatt agctttgaac aattcttatc tcttttcaat agctataaat tatttaataa
    11641 gtaagttaag ggatgcataa actgcatccc ttaacttgtt tttcgtgtac ctattttttg
    11701 tgaat
//
