LOCUS       sequence-223207-        9609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SpCas9 expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9609)
  AUTHORS   Miguel-Escalada I, Bonas-Guarch S, Cebola I, Ponsa-Cobas J, 
            Mendieta-Esteban J, Atla G, Javierre BM, Rolando DMY, Farabella I, 
            Morgan CC, Garcia-Hurtado J, Beucher A, Moran I, Pasquali L, 
            Ramos-Rodriguez M, Appel EVR, Linneberg A, Gjesing AP, Witte DR, 
            Pedersen O, Grarup N, Ravassard P, Torrents D, Mercader JM, Piemonti
            L, Berney T, de Koning EJP, Kerr-Conte J, Pattou F, Fedko IO, Groop 
            L, Prokopenko I, Hansen T, Marti-Renom MA, Fraser P, Ferrer J
  TITLE     Human pancreatic islet three-dimensional chromatin architecture 
            provides insights into the genetics of type 2 diabetes.
  JOURNAL   Nat Genet. 2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 
            10.1038/s41588-019-0457-0.
  PUBMED    31253982
REFERENCE   2  (bases 1 to 9609)
  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 Genet. 
            2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 
            10.1038/s41588-019-0457-0."
COMMENT     Sequence Label: pSpCas9(BB)-T2A-HygR
FEATURES             Location/Qualifiers
     source          1..9609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      111..120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             120..185
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             192..212
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             237..4337
                     /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             237..4337
                     /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             4338..4385
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4401..4454
                     /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             4455..5474
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     primer_bind     complement(5514..5531)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5520..5727
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5738..5878
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5738..5867
                     /label=AAV2 ITR
     rep_origin      5953..6408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6040..6059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6250..6271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6425..6444)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6544..6566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6604..6622)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6690..6794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6795..6863,6864..7655)
                     /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(7013..7032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7826..8414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8315..8334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8476..8724
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8476..8496
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8647..8666
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8743..8818
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8915..9200
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        9202..9479
                     /label=chicken beta-actin promoter
     intron          join(9480..9609,1..99)
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
       61 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg
      121 actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg
      181 ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca
      241 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
      301 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
      361 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
      421 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
      481 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
      541 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
      601 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
      661 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
      721 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
      781 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
      841 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
      901 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
      961 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     1021 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     1081 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     1141 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     1201 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     1261 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     1321 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     1381 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     1441 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     1501 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     1561 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     1621 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     1681 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     1741 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     1801 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     1861 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     1921 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     1981 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     2041 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     2101 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     2161 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     2221 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     2281 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     2341 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     2401 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     2461 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     2521 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     2581 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     2641 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     2701 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     2761 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     2821 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     2881 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     2941 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     3001 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     3061 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     3121 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     3181 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     3241 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     3301 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     3361 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     3421 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     3481 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     3541 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     3601 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     3661 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     3721 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     3781 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     3841 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     3901 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     3961 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     4021 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     4081 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     4141 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     4201 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     4261 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     4321 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     4381 aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     4441 agaatcctgg cccaatgaaa aagcctgaac tcaccgctac ctctgtcgag aagtttctga
     4501 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc cgagggcgaa gaatctcggg
     4561 ctttcagctt cgatgtggga gggcgtggat atgtcctgcg ggtgaatagc tgcgccgatg
     4621 gtttctacaa agatcgctat gtttatcggc actttgcatc cgccgctctc cctattcccg
     4681 aagtgcttga cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     4741 agggtgtcac cttgcaagac ctgcctgaaa ccgaactgcc cgctgttctc cagcccgtcg
     4801 ccgaggccat ggatgccatc gctgccgccg atcttagcca gaccagcggg ttcggcccat
     4861 tcggacctca aggaatcggt caatacacta catggcgcga tttcatctgc gctattgctg
     4921 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcc tccgtcgccc
     4981 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     5041 acgccgattt cggctccaac aatgtcctga ccgacaatgg ccgcataaca gccgtcattg
     5101 actggagcga ggccatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     5161 ggccctggtt ggcttgtatg gagcagcaga cccgctactt cgagcggagg catcccgagc
     5221 ttgcaggatc tcctcggctc cgggcttata tgctccgcat tggtcttgac caactctatc
     5281 agagcttggt tgacggcaat ttcgatgatg cagcttgggc tcagggtcgc tgcgacgcaa
     5341 tcgtccggtc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgctgccg
     5401 tctggaccga tggctgtgtg gaagtgctcg ccgatagtgg aaacagacgc cccagcactc
     5461 gtcctagggc aaaggatctg cagtaagaat tctaactaga gctcgctgat cagcctcgac
     5521 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5581 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     5641 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5701 ggaagagaat agcaggcatg ctggggagcg cggccgcagg aacccctagt gatggagttg
     5761 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     5821 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     5881 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5941 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6001 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6061 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6121 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     6181 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6241 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     6301 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6361 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     6421 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6481 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6541 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6601 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6661 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6721 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6781 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6841 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6901 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6961 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7021 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7081 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     7141 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7201 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7261 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7321 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     7381 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7441 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7501 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7561 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7621 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7681 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7741 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7801 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7861 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7921 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     7981 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8041 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8101 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8161 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     8221 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     8281 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8341 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8401 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8461 cttttgctca catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac
     8521 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     8581 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     8641 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     8701 atcttgtgga aaggacgaaa caccgggtct tcgagaagac ctgttttaga gctagaaata
     8761 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     8821 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
     8881 gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc
     8941 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
     9001 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     9061 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     9121 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     9181 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     9241 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     9301 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
     9361 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     9421 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     9481 gagtcgctgc gacgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
     9541 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
     9601 ccgggctgt
//
