LOCUS       sequence-209389-        9201 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Knock out of EB3 in human cells by CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9201)
  AUTHORS   van Haren J, Charafeddine RA, Ettinger A, Wang H, Hahn KM, Wittmann 
            T
  TITLE     Local control of intracellular microtubule dynamics by EB1 
            photodissociation.
  JOURNAL   Nat Cell Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 
            10.1038/s41556-017-0028-5.
  PUBMED    29379139
REFERENCE   2  (bases 1 to 9201)
  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 Cell 
            Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 
            10.1038/s41556-017-0028-5."
COMMENT     Sequence Label: pSpCas9_BB_2A-GFP_MAPRE3-gRNA#3
FEATURES             Location/Qualifiers
     source          1..9201
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          73..300
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      312..321
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             321..386
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             393..413
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             438..4538
                     /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             438..4538
                     /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             4539..4586
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4602..4655
                     /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(4656..4658,4659..5369)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4656..4658,4659..5369)
                     /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(4959..4978)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5306..5327
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5397..5414)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5403..5610
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5619..5759
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5619..5748
                     /label=AAV2 ITR
     rep_origin      5834..6289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5921..5940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6131..6152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6306..6325)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6425..6447
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6485..6503)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6571..6675
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6676..6744,6745..7536)
                     /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(6894..6913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7707..8295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8196..8215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8357..8605
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8357..8377
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8528..8547
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8627..8702
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8799..9084
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
       61 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      121 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      181 ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg
      241 gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag
      301 gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag gatcatgata
      361 ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc
      421 acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg
      481 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
      541 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
      601 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
      661 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
      721 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
      781 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
      841 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
      901 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
      961 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     1021 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     1081 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     1141 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     1201 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     1261 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     1321 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     1381 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     1441 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     1501 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     1561 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     1621 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     1681 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     1741 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     1801 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     1861 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     1921 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     1981 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     2041 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     2101 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     2161 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     2221 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     2281 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     2341 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     2401 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     2461 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     2521 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     2581 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     2641 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     2701 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     2761 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     2821 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     2881 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     2941 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     3001 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     3061 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     3121 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     3181 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     3241 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     3301 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     3361 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     3421 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     3481 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     3541 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     3601 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     3661 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     3721 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     3781 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     3841 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     3901 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     3961 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     4021 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     4081 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     4141 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     4201 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     4261 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     4321 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     4381 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     4441 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     4501 acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa
     4561 aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga ggaagtctgc
     4621 taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag gagctgttca
     4681 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4741 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4801 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4861 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4921 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     4981 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     5041 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     5101 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     5161 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     5221 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     5281 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     5341 tcactctcgg catggacgag ctgtacaagg aattctaact agagctcgct gatcagcctc
     5401 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5461 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5521 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5581 ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt
     5641 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     5701 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     5761 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     5821 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5881 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     5941 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6001 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     6061 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6121 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     6181 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     6241 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     6301 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6361 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6421 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6481 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6541 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6601 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6661 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6721 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6781 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6841 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6901 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6961 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7021 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7081 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7141 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7201 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7261 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7321 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7381 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7441 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7501 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7561 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7621 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7681 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7741 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7801 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7861 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7921 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7981 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8041 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8101 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8161 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8221 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8281 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8341 ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
     8401 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     8461 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     8521 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     8581 tatcttgtgg aaaggacgaa acaccgtctt ggtatagttg aggtgcgttt tagagctaga
     8641 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     8701 gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc
     8761 gtgcgccaat tctgcagaca aatggctcta gaggtacccg ttacataact tacggtaaat
     8821 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata
     8881 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     8941 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     9001 gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     9061 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     9121 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     9181 gcgatggggg cggggggggg g
//
