LOCUS       sequence-175015-        9137 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 9137)
  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 9137)
  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-Exon4-1sgRNA
FEATURES             Location/Qualifiers
     source          1..9137
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          2..229
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      241..250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             250..315
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             322..342
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             367..4467
                     /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             367..4467
                     /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             4468..4515
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4531..4584
                     /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(4585..4587,4588..5298)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4585..4587,4588..5298)
                     /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(4888..4907)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5235..5256
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5326..5343)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5332..5539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5548..5688
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5548..5677
                     /label=AAV2 ITR
     rep_origin      5763..6218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5850..5869)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6060..6081
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6235..6254)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6354..6376
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6414..6432)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6500..6604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6605..6673,6674..7465)
                     /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(6823..6842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7636..8224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8125..8144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8286..8534
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8286..8306
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8457..8476
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8556..8631
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8728..9013
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
       61 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      121 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt
      181 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt
      241 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
      301 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
      361 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
      421 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
      481 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
      541 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
      601 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
      661 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
      721 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
      781 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
      841 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
      901 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
      961 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     1021 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     1081 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     1141 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     1201 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     1261 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     1321 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     1381 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     1441 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     1501 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     1561 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     1621 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     1681 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     1741 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     1801 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     1861 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     1921 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     1981 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     2041 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     2101 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     2161 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     2221 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     2281 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     2341 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     2401 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     2461 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     2521 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     2581 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     2641 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     2701 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     2761 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     2821 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     2881 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     2941 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     3001 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     3061 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     3121 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     3181 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     3241 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     3301 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     3361 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     3421 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     3481 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     3541 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     3601 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     3661 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     3721 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     3781 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     3841 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     3901 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     3961 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     4021 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     4081 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     4141 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     4201 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     4261 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     4321 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     4381 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     4441 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag
     4501 gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt
     4561 gacgtcgagg agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg
     4621 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4681 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4741 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4801 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4861 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     4921 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4981 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5041 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5101 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     5161 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     5221 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     5281 atggacgagc tgtacaagga attctaacta gagctcgctg atcagcctcg actgtgcctt
     5341 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5401 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5461 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
     5521 atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct
     5581 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     5641 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     5701 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     5761 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5821 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5881 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5941 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     6001 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6061 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     6121 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     6181 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     6241 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     6301 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6361 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     6421 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     6481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6841 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7141 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7321 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7741 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8281 catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     8341 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     8401 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     8461 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     8521 aaggacgaaa caccgccgtc acgctattct gtgcggtttt agagctagaa atagcaagtt
     8581 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
     8641 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
     8701 ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
     8761 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
     8821 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     8881 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
     8941 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     9001 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     9061 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     9121 gggggggggg ggcgggc
//
