LOCUS       sequence-181405-        8459 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CAG-driven ubiquitous expression of catalytically inactive Cas9 
            fused to KRAB repressor domain. Contains 2A-EGFP reporter. For 
            targeted enhancer inactivation in chicken embryos..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8459)
  AUTHORS   Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, 
            Sauka-Spengler T
  TITLE     Genome and epigenome engineering CRISPR toolkit for in vivo 
            modulation of cis-regulatory interactions and gene expression in the
            chicken embryo.
  JOURNAL   Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 
            10.1242/dev.160333.
  PUBMED    29386245
REFERENCE   2  (bases 1 to 8459)
  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: 
            "Development."; date: "2018-02-23"; volume: "145(4). pii: 
            dev.160333. doi"; pages: " 10.1242/dev.160333"
COMMENT     Sequence Label: pCAG dCas9-KRAB-2A-EGFP
FEATURES             Location/Qualifiers
     source          1..8459
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..66
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             73..93
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(103..129,130..132,133..2616,2617..2622,2623..2688,
                     2689..2691,2692..4206)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /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"
     CDS             join(103..129,130..132,133..2619,2620..2622,2623..4206)
                     /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"
     CDS             103..4206
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4219..4239
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4273..4467
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             4534..4554
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4561..4614
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4621..4623,4624..5334)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4621..4623,4624..5334)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4663..4684)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4924..4943)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5271..5292
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5433..5452)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5498..5553
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5552..5571)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5944..5960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5944..5960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5957..5979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5968..5984
                     /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)."
     promoter        complement(join(5992..5998,5999..6016,6017..6022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6037..6058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6117..6312
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6163..6298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6318..6452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6355..6374)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6409..6428
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6520..6537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6691..7279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6771..6790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7450..8241,8242..8310))
                     /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     8073..8092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8311..8415)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
       61 gacaagatgg cccccaagaa gaagaggaag gtgggccgcg gaatggacaa gaagtactcc
      121 attgggctcg ccatcggcac aaacagcgtc ggctgggccg tcattacgga cgagtacaag
      181 gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac
      241 ctcattggcg ccctcctgtt cgactccggg gaaaccgccg aagccacgcg gctcaaaaga
      301 acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt
      361 agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg
      421 gtggaggagg ataaaaagca cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg
      481 gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact
      541 gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga
      601 cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc
      661 caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt
      721 gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc
      781 gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc
      841 gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg
      901 agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac
      961 gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg
     1021 cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat
     1081 gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag
     1141 tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga
     1201 gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc
     1261 gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac
     1321 aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa
     1381 gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt
     1441 cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact
     1501 cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc
     1561 tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag
     1621 gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag
     1681 gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa
     1741 gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa
     1801 gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc
     1861 ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc
     1921 ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt
     1981 gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa
     2041 gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg
     2101 atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat
     2161 ggatttgcca accggaactt catgcagttg atccatgatg actctctcac ctttaaggag
     2221 gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat
     2281 cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa
     2341 ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag
     2401 aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag
     2461 ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt
     2521 cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag
     2581 gaactggaca tcaatcggct ctccgactac gacgtggatg ccatcgtgcc ccagtctttt
     2641 ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag
     2701 agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg
     2761 ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt
     2821 ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag
     2881 atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat
     2941 gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc
     3001 agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat
     3061 gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct
     3121 gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag
     3181 caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc
     3241 aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac
     3301 ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc
     3361 ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc
     3421 aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg
     3481 gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg
     3541 gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc
     3601 acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga
     3661 tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt
     3721 gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg
     3781 gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa
     3841 gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt
     3901 gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac
     3961 ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca
     4021 gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac
     4081 ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca
     4141 ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt
     4201 ggagacagca gggctgaccc caagaagaag aggaaggtgg ctagcgatgc taagtcactg
     4261 actgcctggt cccggacact ggtgaccttc aaggatgtgt ttgtggactt caccagggag
     4321 gagtggaagc tgctggacac tgctcagcag atcctgtaca gaaatgtgat gctggagaac
     4381 tataagaacc tggtttcctt gggttatcag cttactaagc cagatgtgat cctccggttg
     4441 gagaagggag aagagccctg gctggtggag agagaaattc accaagagac ccatcctgat
     4501 tcagagactg catttgaaat caaatcatca gttccgaaaa agaaacgcaa agttgctagc
     4561 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctggtacc
     4621 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     4681 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     4741 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4801 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4861 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4921 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4981 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     5041 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     5101 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     5161 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     5221 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     5281 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagaccggt
     5341 tgatggccgc aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     5401 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     5461 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     5521 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     5581 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     5641 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     5701 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     5761 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     5821 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     5881 ctcgacctgc accgtcgacc agctggtcga cggtgcaccg tcgaccagct tggcgtaatc
     5941 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6001 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6061 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     6121 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6181 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6241 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6301 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6361 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6421 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     6481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7561 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7741 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7921 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8221 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8281 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8341 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8401 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattg
//
