LOCUS       sequence-175018-        9081 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  CRISPR KO of Trp73.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9081)
  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 9081)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Trp73-Exon4-3sgRNA
FEATURES             Location/Qualifiers
     source          1..9081
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          10..237
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      249..258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             258..323
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             330..350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             375..4475
                     /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             375..4475
                     /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             4476..4523
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4539..4592
                     /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(4593..4595,4596..5306)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4593..4595,4596..5306)
                     /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(4896..4915)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5243..5264
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5334..5351)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5340..5547
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5556..5696
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5556..5685
                     /label=AAV2 ITR
     rep_origin      5771..6226
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5858..5877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6068..6089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6243..6262)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6362..6384
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6422..6440)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6508..6612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6613..6681,6682..7473)
                     /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(6831..6850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7644..8232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8133..8152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8294..8542
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8294..8314
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8465..8484
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8564..8639
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8736..9021
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
       61 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
      121 ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt
      181 ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt
      241 ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat catgatattg
      301 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
      361 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
      421 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
      481 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
      541 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
      601 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
      661 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
      721 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
      781 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
      841 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
      901 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
      961 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     1021 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     1081 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     1141 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     1201 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     1261 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     1321 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     1381 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     1441 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     1501 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     1561 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     1621 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     1681 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     1741 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     1801 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     1861 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     1921 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     1981 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     2041 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     2101 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     2161 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     2221 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     2281 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     2341 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     2401 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     2461 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     2521 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     2581 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     2641 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     2701 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     2761 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     2821 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     2881 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     2941 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     3001 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     3061 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     3121 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     3181 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     3241 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     3301 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     3361 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     3421 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     3481 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     3541 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     3601 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     3661 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     3721 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     3781 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     3841 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     3901 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     3961 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     4021 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     4081 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     4141 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     4201 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     4261 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     4321 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     4381 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     4441 agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg
     4501 ccggccaggc aaaaaagaaa aaggaattcg gcagtggaga gggcagagga agtctgctaa
     4561 catgcggtga cgtcgaggag aatcctggcc cagtgagcaa gggcgaggag ctgttcaccg
     4621 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4681 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4741 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4801 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4861 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4921 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4981 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     5041 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     5101 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     5161 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     5221 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     5281 ctctcggcat ggacgagctg tacaaggaat tctaactaga gctcgctgat cagcctcgac
     5341 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5401 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     5461 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5521 ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc
     5581 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     5641 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     5701 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     5761 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5821 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     5881 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5941 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     6001 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6061 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     6121 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     6181 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     6241 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6301 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6361 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     6421 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6481 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7141 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7321 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8281 tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa
     8341 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
     8401 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
     8461 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
     8521 cttgtggaaa ggacgaaaca ccgccggggt agtcggtatt ggagttttag agctagaaat
     8581 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     8641 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
     8701 cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc
     8761 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
     8821 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8881 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     8941 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     9001 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     9061 tcccccccct ccccaccccc g
//
