LOCUS       sequence-174386-        8898 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 nuclease, FBL-specific gRNA, and the PITCh-gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8898)
  AUTHORS   Sakuma T, Nakade S, Sakane Y, Suzuki KT, Yamamoto T
  TITLE     MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the 
            PITCh systems.
  JOURNAL   Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub
            2015 Dec 17.
  PUBMED    26678082
REFERENCE   2  (bases 1 to 8898)
  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: 
            "10.1038/nprot.2015.140"; journalName: "Nat Protoc"; date: 
            "2016-01"; volume: "11"; issue: "1"; pages: "118-33"
COMMENT     Sequence Label: pX330A-FBL/PITCh
FEATURES             Location/Qualifiers
     source          1..8898
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        119..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(501..576)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(597..845)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        complement(943..1018)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(1040..1288)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     rep_origin      complement(1350..1938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1430..1449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2109..2900,2901..2969))
                     /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     2732..2751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2970..3074)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3142..3160
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3198..3220)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3320..3339
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3356..3811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3493..3514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3705..3724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3886..4026
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3897..4026
                     /label=AAV2 ITR
     polyA_signal    4035..4242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4231..4248
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4276..4323)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4324..8424)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4324..8424)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(8449..8469)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8476..8541)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      8541..8550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          8562..8789
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
       61 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
      121 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
      181 tgtactgggc acaatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
      241 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
      301 attgacgtca atggaaagtc cctattggcg ttactattga cgtcaatggg cgggggtcgt
      361 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgggtacc tctagagcca
      421 tttgtctgca gaattggcgc acgcgctaaa aacggactag ccttatttta acttgctatt
      481 tctagctcta aaacaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact
      541 agccttattt taacttgcta tttctagctc taaaacaaca cgtacgcgta cgatgcggtg
      601 tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca agttacggta
      661 agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat
      721 tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcca
      781 attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg
      841 ccctcacatg cctctagagc catttgtctg cagaattggc gcacgcgcta aaaacggact
      901 agccttattt taacttgcta tttctagctc taaaacaaaa aagcaccgac tcggtgccac
      961 tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaactg
     1021 gggggtggcc tgtgagagcg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat
     1081 cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa
     1141 ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg
     1201 tgtttacagt caaattaatt ccaattatct ctctaacagc cttgtatcgt atatgcaaat
     1261 atgaaggaat catgggaaat aggccctcac atgtgagcaa aaggccagca aaaggccagg
     1321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2041 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2101 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2161 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2221 tctggcccca gtgctgcaat gataccgcgg cttccacgct caccggctcc agatttatca
     2281 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2341 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2401 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2461 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2521 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2581 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2641 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2701 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2761 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2821 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2881 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2941 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3001 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3061 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     3121 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     3181 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     3241 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     3301 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt
     3361 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3421 aaccaataga ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag
     3481 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3541 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca
     3601 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3661 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3721 ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3781 gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg
     3841 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc
     3901 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
     3961 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     4021 gttcctgcgg ccgctcccca gcatgcctgc tattctcttc ccaatcctcc cccttgctgt
     4081 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     4141 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     4201 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcgag
     4261 ctctaggaat tcttactttt tcttttttgc ctggccggcc tttttcgtgg ccgccggcct
     4321 tttgtcgcct cccagctgag acaggtcgat ccgtgtctcg tacaggccgg tgatgctctg
     4381 gtggatcagg gtggcgtcca gcacctcttt ggtgctggtg tacctcttcc ggtcgatggt
     4441 ggtgtcaaag tacttgaagg cggcaggggc tcccagattg gtcagggtaa acaggtggat
     4501 gatattctcg gcctgctctc tgatgggctt atcccggtgc ttgttgtagg cggacagcac
     4561 tttgtccaga ttagcgtcgg ccaggatcac tctcttggag aactcgctga tctgctcgat
     4621 gatctcgtcc aggtagtgct tgtgctgttc cacaaacagc tgtttctgct cattatcctc
     4681 gggggagccc ttcagcttct catagtggct ggccaggtac aggaagttca catatttgga
     4741 gggcagggcc agttcgtttc ccttctgcag ttcgccggca gaggccagca ttctcttccg
     4801 gccgttttcc agctcgaaca gggagtactt aggcagcttg atgatcaggt cctttttcac
     4861 ttctttgtag cccttggctt ccagaaagtc gatgggattc ttctcgaagc tgcttctttc
     4921 catgatggtg atccccagca gctctttcac actcttcagt ttcttggact tgcccttttc
     4981 cactttggcc accaccagca cagaataggc cacggtgggg ctgtcgaagc cgccgtactt
     5041 cttagggtcc cagtccttct ttctggcgat cagcttatcg ctgttcctct tgggcaggat
     5101 agactctttg ctgaagccgc ctgtctgcac ctcggtcttt ttcacgatat tcacttgggg
     5161 catgctcagc actttccgca cggtggcaaa atcccggccc ttatcccaca cgatctcccc
     5221 ggtttcgccg tttgtctcga tcagaggccg cttccggatc tcgccgttgg ccagggtaat
     5281 ctcggtcttg aaaaagttca tgatgttgct gtagaagaag tacttggcgg tagccttgcc
     5341 gatttcctgc tcgctcttgg cgatcatctt ccgcacgtcg tacaccttgt agtcgccgta
     5401 cacgaactcg ctttccagct tagggtactt tttgatcagg gcggttccca cgacggcgtt
     5461 caggtaggcg tcgtgggcgt ggtggtagtt gttgatctcg cgcactttgt aaaactggaa
     5521 atccttccgg aaatcggaca ccagcttgga cttcagggtg atcactttca cttcccggat
     5581 cagcttgtca ttctcgtcgt acttagtgtt catccgggag tccaggatct gtgccacgtg
     5641 ctttgtgatc tgccgggttt ccaccagctg tctcttgatg aagccggcct tatccagttc
     5701 gctcaggccg cctctctcgg ccttggtcag attgtcgaac tttctctggg taatcagctt
     5761 ggcgttcagc agctgccgcc agtagttctt catcttcttc acgacctctt cggagggcac
     5821 gttgtcgctc ttgccccggt tcttgtcgct tctggtcagc accttgttgt cgatggagtc
     5881 gtccttcaga aagctctgag gcacgatatg gtccacatcg tagtcggaca gccggttgat
     5941 gtccagttcc tggtccacgt acatatcccg cccattctgc aggtagtaca ggtacagctt
     6001 ctcgttctgc agctgggtgt tttccacggg gtgttctttc aggatctggc tgcccagctc
     6061 tttgatgccc tcttcgatcc gcttcattct ctcgcggctg ttcttctgtc ccttctgggt
     6121 ggtctggttc tctctggcca tttcgatcac gatgttctcg ggcttgtgcc ggcccatcac
     6181 tttcacgagc tcgtccacca ccttcactgt ctgcaggatg cccttcttaa tggcggggct
     6241 gccggccaga ttggcaatgt gctcgtgcag gctatcgccc tggccggaca cctgggcttt
     6301 ctggatgtcc tctttaaagg tcaggctgtc gtcgtggatc agctgcatga agtttctgtt
     6361 ggcgaagccg tcggacttca ggaaatccag gattgtcttg ccggactgct tgtcccggat
     6421 gccgttgatc agcttccggc tcagcctgcc ccagccggtg tatctccgcc gcttcagctg
     6481 cttcatcact ttgtcgtcga acaggtgggc ataggttttc agccgttcct cgatcatctc
     6541 tctgtcctca aacagtgtca gggtcagcac gatatcttcc agaatgtcct cgttttcctc
     6601 attgtccagg aagtccttgt ccttgataat tttcagcaga tcgtggtatg tgcccaggga
     6661 ggcgttgaac cgatcttcca cgccggagat ttccacggag tcgaagcact cgattttctt
     6721 gaagtagtcc tctttcagct gcttcacggt cactttccgg ttggtcttga acagcaggtc
     6781 cacgatggcc tttttctgct cgccgctcag gaaggcgggc tttctcattc cctcggtcac
     6841 gtatttcact ttggtcagct cgttatacac ggtgaagtac tcgtacagca ggctgtgctt
     6901 gggcagcacc ttctcgttgg gcaggttctt atcgaagttg gtcatccgct cgatgaagct
     6961 ctgggcggaa gcgcccttgt ccaccacttc ctcgaagttc cagggggtga tggtttcctc
     7021 gctctttctg gtcatccagg cgaatctgct gtttcccctg gccagagggc ccacgtagta
     7081 ggggatgcgg aaggtcagga tcttctcgat cttttcccgg ttgtccttca ggaatgggta
     7141 aaaatcttcc tgccgccgca gaatggcgtg cagctctccc aggtggatct ggtgggggat
     7201 gctgccgttg tcgaaggtcc gctgcttccg cagcaggtcc tctctgttca gcttcacgag
     7261 cagttcctcg gtgccgtcca tcttttccag gatgggcttg atgaacttgt agaactcttc
     7321 ctggctggct ccgccgtcaa tgtagccggc gtagccgttc ttgctctggt cgaagaaaat
     7381 ctctttgtac ttctcaggca gctgctgccg cacgagagct ttcagcaggg tcaggtcctg
     7441 gtggtgctcg tcgtatctct tgatcataga ggcgctcagg ggggccttgg tgatctcggt
     7501 gttcactctc aggatgtcgc tcagcaggat ggcgtcggac aggttcttgg cggccagaaa
     7561 caggtcggcg tactggtcgc cgatctgggc cagcaggttg tccaggtcgt cgtcgtaggt
     7621 gtccttgctc agctgcagtt tggcatcctc ggccaggtcg aagttgctct tgaagttggg
     7681 ggtcaggccc aggctcaggg caatcaggtt tccgaacagg ccattcttct tctcgccggg
     7741 cagctgggcg atcagatttt ccagccgtct gctcttgctc agtctggcag acaggatggc
     7801 cttggcgtcc acgccgctgg cgttgatggg gttttcctcg aacagctggt tgtaggtctg
     7861 caccagctgg atgaacagct tgtccacgtc gctgttgtcg gggttcaggt cgccctcgat
     7921 caggaagtgg ccccggaact tgatcatgtg ggccagggcc agatagatca gccgcaggtc
     7981 ggccttgtcg gtgctgtcca ccagtttctt tctcaggtgg tagatggtgg ggtacttctc
     8041 gtggtaggcc acctcgtcca cgatgttgcc gaagatgggg tgccgctcgt gcttcttatc
     8101 ctcttccacc aggaaggact cttccagtct gtggaagaag ctgtcgtcca ccttggccat
     8161 ctcgttgctg aagatctctt gcagatagca gatccggttc ttccgtctgg tgtatcttct
     8221 tctggcggtt ctcttcagcc gggtggcctc ggctgtttcg ccgctgtcga acagcagggc
     8281 tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg cccagcacct tgaatttctt
     8341 gctgggcacc ttgtactcgt cggtgatcac ggcccagccc acagagttgg tgccgatgtc
     8401 caggccgatg ctgtacttct tgtcggctgc tgggactccg tggataccga ccttccgctt
     8461 cttctttggg gccatcttat cgtcatcgtc tttgtaatca atatcatgat ccttgtagtc
     8521 tccgtcgtgg tccttatagt ccatggtggc accggtccaa cctgaaaaaa agtgatttca
     8581 ggcaggtgct ccaggtaatt aaacattaat accccaccaa ccaaccatcc cttaaaccct
     8641 tacctcttgc tcagctaatt acagcccgga ggagaagggc cgtcccgccc gctcacctgt
     8701 gggagtaacg cggtcagtca gagccggggc gggcggcgcg aggcggcggc ggagcggggc
     8761 acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg cgcttcgctt tttatagggc
     8821 cgccgccgcc gccgcctcgc cataaaagga aactttcgga gcgcgccgct ctgattggct
     8881 gccgccgcac ctctccgc
//
