LOCUS       sequence-175016-        9090 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR KO of Trp63.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9090)
  AUTHORS   Wang Q, Zou Y, Nowotschin S, Kim SY, Li QV, Soh CL, Su J, Zhang C, 
            Shu W, Xi Q, Huangfu D, Hadjantonakis AK, Massague J
  TITLE     The p53 Family Coordinates Wnt and Nodal Inputs in Mesendodermal 
            Differentiation of Embryonic Stem Cells.
  JOURNAL   Cell Stem Cell. 2017 Jan 5;20(1):70-86. doi: 
            10.1016/j.stem.2016.10.002. Epub 2016 Nov 23.
  PUBMED    27889317
REFERENCE   2  (bases 1 to 9090)
  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: 
            "10.1016/j.stem.2016.10.002"; journalName: "Cell Stem Cell"; date: 
            "2017-01-5- 5"; volume: "20"; issue: "1"; pages: "70-86"
COMMENT     Sequence Label: Px458-Cas9n-Trp63-Exon4-2sgRNA
FEATURES             Location/Qualifiers
     source          1..9090
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      223..232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             232..297
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             304..324
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             349..4449
                     /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             349..4449
                     /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             4450..4497
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4513..4566
                     /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(4567..4569,4570..5280)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4567..4569,4570..5280)
                     /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(4870..4889)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5217..5238
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5308..5325)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5314..5521
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5530..5670
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5530..5659
                     /label=AAV2 ITR
     rep_origin      5745..6200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5832..5851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6042..6063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6217..6236)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6336..6358
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6396..6414)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6482..6586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6587..6655,6656..7447)
                     /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(6805..6824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7618..8206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8107..8126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8268..8516
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8268..8288
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8439..8458
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8538..8613
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8710..8995
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
       61 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct
      121 gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct
      181 ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag
      241 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
      301 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac
      361 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
      421 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
      481 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
      541 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
      601 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
      661 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
      721 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
      781 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
      841 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
      901 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
      961 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     1021 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     1081 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     1141 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     1201 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     1261 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     1321 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     1381 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     1441 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     1501 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     1561 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     1621 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     1681 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     1741 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     1801 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     1861 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     1921 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     1981 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     2041 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     2101 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     2161 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     2221 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     2281 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     2341 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     2401 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     2461 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     2521 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     2581 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     2641 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     2701 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     2761 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     2821 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     2881 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     2941 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     3001 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     3061 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     3121 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     3181 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     3241 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     3301 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     3361 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     3421 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     3481 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     3541 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     3601 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     3661 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     3721 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     3781 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     3841 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     3901 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     3961 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     4021 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     4081 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     4141 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     4201 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     4261 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     4321 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     4381 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     4441 ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggaa
     4501 ttcggcagtg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct
     4561 ggcccagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4621 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4681 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4741 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4801 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4861 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4921 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4981 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     5041 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     5101 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     5161 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     5221 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5281 gaattctaac tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     5341 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     5401 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     5461 gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg
     5521 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     5581 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     5641 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     5701 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     5761 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5821 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5881 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5941 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6001 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6061 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     6121 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6181 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     6241 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6301 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6361 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6421 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7261 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc
     7321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     8041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     8101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     8221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag ggcctatttc
     8281 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta
     8341 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
     8401 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
     8461 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgtgg
     8521 gcccgggtaa tctgtgtgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     8581 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca
     8641 agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct
     8701 agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     8761 ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8821 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     8881 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt
     8941 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     9001 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     9061 gtatttattt attttttaat tattttgtgc
//
