LOCUS       sequence-182296-        9150 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR-GFP-gRNA for cutting WT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9150)
  AUTHORS   Bao X, Lian X, Hacker TA, Schmuck EG, Qian T, Bhute VJ, Han T, Shi 
            M, Drowley L, Plowright A, Wang QD, Goumans MJ, Palecek SP
  TITLE     Long-term self-renewing human epicardial cells generated from 
            pluripotent stem cells under defined xeno-free conditions.
  JOURNAL   Nat Biomed Eng. 2016;1. pii: 0003. doi: 10.1038/s41551-016-0003. 
            Epub 2016 Dec 5.
  PUBMED    28462012
REFERENCE   2  (bases 1 to 9150)
  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: "Nat Biomed 
            Eng."; date: "2016"; volume: "1. pii: 0003. doi"; pages: " 
            10.1038/s41551-016-0003. Epub 2016 Dec 5"
COMMENT     Sequence Label: WT1 KI gRNA2
FEATURES             Location/Qualifiers
     source          1..9150
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      228..237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             237..302
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             309..329
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             354..4454
                     /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             354..4454
                     /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             4455..4502
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4518..4571
                     /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(4572..4574,4575..5285)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4572..4574,4575..5285)
                     /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(4875..4894)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5222..5243
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5313..5330)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5319..5526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5535..5675
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5535..5664
                     /label=AAV2 ITR
     rep_origin      5750..6205
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5837..5856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6047..6068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6222..6241)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6341..6363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6401..6419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6487..6591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6592..6660,6661..7452)
                     /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(6810..6829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7623..8211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8112..8131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8273..8521
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8273..8293
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8444..8463
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8542..8617
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8714..8999
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
       61 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      121 tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat
      181 tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc accatggact
      241 ataaggacca cgacggagac tacaaggatc atgatattga ttacaaagac gatgacgata
      301 agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga
      361 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
      421 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
      481 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
      541 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
      601 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
      661 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
      721 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
      781 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
      841 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
      901 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
      961 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     1021 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
     1081 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     1141 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     1201 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     1261 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     1321 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     1381 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     1441 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     1501 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     1561 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     1621 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     1681 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     1741 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     1801 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     1861 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     1921 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     1981 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     2041 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     2101 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     2161 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     2221 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     2281 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     2341 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     2401 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     2461 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     2521 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     2581 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     2641 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     2701 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     2761 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     2821 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     2881 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     2941 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     3001 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     3061 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     3121 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     3181 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     3241 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     3301 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     3361 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     3421 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     3481 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     3541 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     3601 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     3661 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     3721 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     3781 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     3841 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     3901 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     3961 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     4021 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     4081 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     4141 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     4201 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     4261 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     4321 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     4381 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     4441 agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa
     4501 aggaattcgg cagtggagag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga
     4561 atcctggccc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     4621 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4681 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4741 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4801 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4861 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4921 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4981 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     5041 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     5101 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     5161 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     5221 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     5281 acaaggaatt ctaactagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     5341 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     5401 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     5461 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     5521 tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     5581 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     5641 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     5701 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     5761 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5821 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5881 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5941 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     6001 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6061 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     6121 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6181 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     6241 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6301 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     6361 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     6421 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     6481 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6541 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6601 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6661 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6721 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6781 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6841 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6901 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6961 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7021 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7081 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7141 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7201 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7261 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
     7321 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7381 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7441 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7501 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7561 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7621 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7681 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7741 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7801 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7861 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7921 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7981 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8041 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8101 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8161 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8221 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtgagggcct
     8281 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
     8341 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     8401 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     8461 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     8521 cggacactga acggtccccg agttttagag ctagaaatag caagttaaaa taaggctagt
     8581 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag agctagaaat
     8641 agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc agacaaatgg
     8701 ctctagaggt acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     8761 acccccgccc attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg
     8821 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     8881 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga
     8941 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     9001 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
     9061 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
     9121 ggcgcgcgcc aggcggggcg gggcggggcg
//
