| 质粒名称: | pEF1a-FB-dCas9-puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 100547 |
| 存储实验室: | Jian Xu |
|
载体骨架基本信息 |
|
| 载体名称: | pEF1a-FB-puro |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7064 bp |
| 完整质粒大小: | 11244 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | N-terminal Flag and biotin-acceptor-site (FB)-tagged dCas9 |
| 物种来源: | -- |
| 插入大小: | 4305 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEF1a-FB-dCas9-puro was a gift from Jian Xu (Addgene plasmid # 100547 ; http://n2t.net/addgene:100547 ; RRID:Addgene_100547) |
| 参考文献部分: | In Situ Capture of Chromatin Interactions by Biotinylated dCas9. Liu X, Zhang Y, Chen Y, Li M, Zhou F, Li K, Cao H, Ni M, Liu Y, Gu Z, Dickerson KE, Xie S, Hon GC, Xuan Z, Zhang MQ, Shao Z, Xu J. Cell. 2017 Aug 24;170(5):1028-1043.e19. doi: 10.1016/j.cell.2017.08.003. 10.1016/j.cell.2017.08.003 PubMed 28841410 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pEF1a-FB-dCas9-puro 11229 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses N-terminal Flag and biotin-acceptor-site (FB)-tagged dCas9
protein.
ACCESSION .
VERSION .
KEYWORDS pEF1a-FB-dCas9-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11229)
AUTHORS Liu X, Zhang Y, Chen Y, Li M, Zhou F, Li K, Cao H, Ni M, Liu Y, Gu
Z, Dickerson KE, Xie S, Hon GC, Xuan Z, Zhang MQ, Shao Z, Xu J
TITLE In Situ Capture of Chromatin Interactions by Biotinylated dCas9.
JOURNAL Cell. 2017 Aug 24;170(5):1028-1043.e19. doi:
10.1016/j.cell.2017.08.003.
PUBMED 28841410
REFERENCE 2 (bases 1 to 11229)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11229
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 156..179
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
protein_bind 279..300
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 315..345
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 353..369
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 377..393
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 425..635
/label=SV40 promoter
/note="SV40 early promoter"
promoter 447..643
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 494..629
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 556..575
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 730..1908
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 961..1899
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1856..1876
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 1941..1964
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2004..2048
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 2079..2099
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2100..6200
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 6210..6230
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(6961..6977)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7059..7075)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7060..7076)
/label=pBluescriptKS
/note="For pBluescript vector"
promoter complement(7104..7122)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(7143..7159)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 7167..7183
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(7191..7221)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7236..7257
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7374..7391)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7545..8133)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7625..7644)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8304..9164)
/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 8927..8946
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9165..9269)
/gene="bla"
/label=AmpR promoter
rep_origin 9296..9751
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9383..9402)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 9593..9614
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 9892..9908
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9915..9934
/label=T7
/note="T7 promoter, forward primer"
promoter 9915..9933
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 10028..10252
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 10241..10258
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
CDS complement(10333..10932)
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(10416..10436)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 10913..10932
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
promoter complement(10944..214)
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(10992..11011)
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
ORIGIN
1 cacgacgcga gctgcggggc gggggggaac ttcctgacta ggggaggagt agaaggtggc
61 gcgaaggggc caccaaagaa cggagccggt tggcgcctac cggtggatgt ggaatgtgtg
121 cgaggccaga ggccacttgt gtagcgccaa gtgcccagcg gggctgctaa agcgcatgct
181 ccagactgcc ttgggaaaag cgcctcccct acccggtaga attcgatctg gcacgacagg
241 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
301 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
361 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct taattccctc
421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
481 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
541 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
601 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttt gcaaagatgg
661 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
721 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
781 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
841 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
901 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
961 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
1021 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
1081 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
1141 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
1201 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
1261 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
1321 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
1381 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
1441 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
1501 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
1561 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
1621 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
1681 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
1741 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
1801 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
1861 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg aattgatcca
1921 gatctttcga aatcaccatg gactacaaag acgatgacga taaagcaaga agagcatctg
1981 tggaattcct gcagcccggg tccggcctga acgacatctt cgaggctcag aaaatcgaat
2041 ggcacgaagg cgcgccgagc tcgaggatcc ttgctagccc caaaaagaag aggaaagtgg
2101 acaagaagta ttctatcgga ctggccatcg ggactaatag cgtcgggtgg gccgtgatca
2161 ctgacgagta caaggtgccc tctaagaagt tcaaggtgct cgggaacacc gaccggcatt
2221 ccatcaagaa aaatctgatc ggagctctcc tctttgattc aggggagacc gctgaagcaa
2281 cccgcctcaa gcggactgct agacggcggt acaccaggag gaagaaccgg atttgttacc
2341 ttcaagagat attctccaac gaaatggcaa aggtcgacga cagcttcttc cataggctgg
2401 aagaatcatt cctcgtggaa gaggataaga agcatgaacg gcatcccatc ttcggtaata
2461 tcgtcgacga ggtggcctat cacgagaaat acccaaccat ctaccatctt cgcaaaaagc
2521 tggtggactc aaccgacaag gcagacctcc ggcttatcta cctggccctg gcccacatga
2581 tcaagttcag aggccacttc ctgatcgagg gcgacctcaa tcctgacaat agcgatgtgg
2641 ataaactgtt catccagctg gtgcagactt acaaccagct ctttgaagag aaccccatca
2701 atgcaagcgg agtcgatgcc aaggccattc tgtcagcccg gctgtcaaag agccgcagac
2761 ttgagaatct tatcgctcag ctgccgggtg aaaagaaaaa tggactgttc gggaacctga
2821 ttgctctttc acttgggctg actcccaatt tcaagtctaa tttcgacctg gcagaggatg
2881 ccaagctgca actgtccaag gacacctatg atgacgatct cgacaacctc ctggcccaga
2941 tcggtgacca atacgccgac cttttccttg ctgctaagaa tctttctgac gccatcctgc
3001 tgtctgacat tctccgcgtg aacactgaaa tcaccaaggc ccctctttca gcttcaatga
3061 ttaagcggta tgatgagcac caccaggacc tgaccctgct taaggcactc gtccggcagc
3121 agcttccgga gaagtacaag gaaatcttct ttgaccagtc aaagaatgga tacgccggct
3181 acatcgacgg aggtgcctcc caagaggaat tttataagtt tatcaaacct atccttgaga
3241 agatggacgg caccgaagag ctcctcgtga aactgaatcg ggaggatctg ctgcggaagc
3301 agcgcacttt cgacaatggg agcattcccc accagatcca tcttggggag cttcacgcca
3361 tccttcggcg ccaagaggac ttctacccct ttcttaagga caacagggag aagattgaga
3421 aaattctcac tttccgcatc ccctactacg tgggacccct cgccagagga aatagccggt
3481 ttgcttggat gaccagaaag tcagaagaaa ctatcactcc ctggaacttc gaagaggtgg
3541 tggacaaggg agccagcgct cagtcattca tcgaacggat gactaacttc gataagaacc
3601 tccccaatga gaaggtcctg ccgaaacatt ccctgctcta cgagtacttt accgtgtaca
3661 acgagctgac caaggtgaaa tatgtcaccg aagggatgag gaagcccgca ttcctgtcag
3721 gcgaacaaaa gaaggcaatt gtggaccttc tgttcaagac caatagaaag gtgaccgtga
3781 agcagctgaa ggaggactat ttcaagaaaa ttgaatgctt cgactctgtg gagattagcg
3841 gggtcgaaga tcggttcaac gcaagcctgg gtacctacca tgatctgctt aagatcatca
3901 aggacaagga ttttctggac aatgaggaga acgaggacat ccttgaggac attgtcctga
3961 ctctcactct gttcgaggac cgggaaatga tcgaggagag gcttaagacc tacgcccatc
4021 tgttcgacga taaagtgatg aagcaactta aacggagaag atataccgga tggggacgcc
4081 ttagccgcaa actcatcaac ggaatccggg acaaacagag cggaaagacc attcttgatt
4141 tccttaagag cgacggattc gctaatcgca acttcatgca acttatccat gatgattccc
4201 tgacctttaa ggaggacatc cagaaggccc aagtgtctgg acaaggtgac tcactgcacg
4261 agcatatcgc aaatctggct ggttcacccg ctattaagaa gggtattctc cagaccgtga
4321 aagtcgtgga cgagctggtc aaggtgatgg gtcgccataa accagagaac attgtcatcg
4381 agatggccag ggaaaaccag actacccaga agggacagaa gaacagcagg gagcggatga
4441 aaagaattga ggaagggatt aaggagctcg ggtcacagat ccttaaagag cacccggtgg
4501 aaaacaccca gcttcagaat gagaagctct atctgtacta ccttcaaaat ggacgcgata
4561 tgtatgtgga ccaagagctt gatatcaaca ggctctcaga ctacgacgtg gacgccatcg
4621 tccctcagag cttcctcaaa gacgactcaa ttgacaataa ggtgctgact cgctcagaca
4681 agaaccgggg aaagtcagat aacgtgccct cagaggaagt cgtgaaaaag atgaagaact
4741 attggcgcca gcttctgaac gcaaagctga tcactcagcg gaagttcgac aatctcacta
4801 aggctgagag gggcggactg agcgaactgg acaaagcagg attcattaaa cggcaacttg
4861 tggagactcg gcagattact aaacatgtcg cccaaatcct tgactcacgc atgaatacca
4921 agtacgacga aaacgacaaa cttatccgcg aggtgaaggt gattaccctg aagtccaagc
4981 tggtcagcga tttcagaaag gactttcaat tctacaaagt gcgggagatc aataactatc
5041 atcatgctca tgacgcatat ctgaatgccg tggtgggaac cgccctgatc aagaagtacc
5101 caaagctgga aagcgagttc gtgtacggag actacaaggt ctacgacgtg cgcaagatga
5161 ttgccaaatc tgagcaggag atcggaaagg ccaccgcaaa gtacttcttc tacagcaaca
5221 tcatgaattt cttcaagacc gaaatcaccc ttgcaaacgg tgagatccgg aagaggccgc
5281 tcatcgagac taatggggag actggcgaaa tcgtgtggga caagggcaga gatttcgcta
5341 ccgtgcgcaa agtgctttct atgcctcaag tgaacatcgt gaagaaaacc gaggtgcaaa
5401 ccggaggctt ttctaaggaa tcaatcctcc ccaagcgcaa ctccgacaag ctcattgcaa
5461 ggaagaagga ttgggaccct aagaagtacg gcggattcga ttcaccaact gtggcttatt
5521 ctgtcctggt cgtggctaag gtggaaaaag gaaagtctaa gaagctcaag agcgtgaagg
5581 aactgctggg tatcaccatt atggagcgca gctccttcga gaagaaccca attgactttc
5641 tcgaagccaa aggttacaag gaagtcaaga aggaccttat catcaagctc ccaaagtata
5701 gcctgttcga actggagaat gggcggaagc ggatgctcgc ctccgctggc gaacttcaga
5761 agggtaatga gctggctctc ccctccaagt acgtgaattt cctctacctt gcaagccatt
5821 acgagaagct gaaggggagc cccgaggaca acgagcaaaa gcaactgttt gtggagcagc
5881 ataagcatta tctggacgag atcattgagc agatttccga gttttctaaa cgcgtcattc
5941 tcgctgatgc caacctcgat aaagtcctta gcgcatacaa taagcacaga gacaaaccaa
6001 ttcgggagca ggctgagaat atcatccacc tgttcaccct caccaatctt ggtgcccctg
6061 ccgcattcaa gtacttcgac accaccatcg accggaaacg ctatacctcc accaaagaag
6121 tgctggacgc caccctcatc caccagagca tcaccggact ttacgaaact cggattgacc
6181 tctcacagct cggaggggat gagggagctc ccaagaaaaa gcgcaaggta ggtagttcct
6241 aatctagatg agggtcccca cctgggaccc ttgagagtat caggtctccc acgtgggaga
6301 caagaaatcc ctgtttaata tttaaacagc agtgttcccc atctgggtcc ttgcacccct
6361 cactctggcc tcagccgact gcacagcggc ccctgcatcc ccttggctgt gaggcccctg
6421 gacaagcaga ggtggccaga gctgggaggc atggccctgg ggtcccacga atttgctggg
6481 gaatctcgtt tttcttctta agacttttgg gacatggttt gactcccgaa catcaccgac
6541 gtgtctcctg tttttctggg tggcctcggg acacctgccc tgcccccacg agggtcagga
6601 ctgtgactct ttttagggcc aggcaggtgc ctggacattt gccttgctgg acggggactg
6661 gggatgtggg agggagcaga caggaggaat catgtcaggc ctgtgtgtga aaggaagctc
6721 cactgtcacc ctccacctct tcacccccca ctcaccagtg tcccctccac tgtcacattg
6781 taactgaact tcaggataat aaagtgtttg cctccaaaaa aaaaaaaaaa aaaaaaaaaa
6841 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
6901 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaccgaattc
6961 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
7021 ccttgcagca catccccctt tcgccagatc aagcttatcg ataccgtcga cctcgagggg
7081 gggcccggta ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca
7141 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
7201 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
7261 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
7321 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
7381 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
7441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
7501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
7561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
7621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
7681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
7741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
7801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
7861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
7921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
7981 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
8041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
8101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
8161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
8221 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
8281 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
8341 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
8401 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
8461 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
8521 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
8581 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
8641 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
8701 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
8761 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
8821 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
8881 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
8941 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
9001 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
9061 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
9121 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
9181 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
9241 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
9301 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
9361 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
9421 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
9481 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
9541 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
9601 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
9661 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
9721 aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt
9781 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
9841 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
9901 cggccagtga attgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg
9961 gccgctctag cctcgaggct agaactagtg gatctcgagc cccagctggt tctttccgcc
10021 tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
10081 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
10141 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
10201 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
10261 gatcagcgag ctctacgatg catggggtcg tgcgctcctt tcggtcgggc gctgcgggtc
10321 gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg
10381 cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg
10441 cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag
10501 cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag
10561 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
10621 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
10681 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
10741 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
10801 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
10861 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
10921 gtactcggtc atggtaagct ggtcgaaagg cccggagatg aggaagagga gaacagcgcg
10981 gcagacgtgc gcttttgaag cgtgcagaat gccgggcctc cggaggacct tcgggcgccc
11041 gccccgcccc tgagcccgcc cctgagcccg cccccggacc caccccttcc cagcctctga
11101 gcccagaaag cgaaggagca aagctgctat tggccgctgc cccaaaggcc tacccgcttc
11161 cattgctcag cggtgctgtc catctgcacg agactagtga gacgtgctac ttccatttgt
11221 cacgtcctg
//
