| 质粒名称: | pX330A_FokI-1x3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 63603 |
| 存储实验室: | Takashi Yamamoto |
|
载体骨架基本信息 |
|
| 载体名称: | pUC ori vector |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达,CRISPR |
| 筛选标记: | -- |
| 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: | FokI-dCas9 |
| 物种来源: | S. pyogenes and F. okeanokoites |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CBh |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 (TAATACGACTCACTATAGGG) |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pX330A_FokI-1x3 was a gift from Takashi Yamamoto (Addgene plasmid # 63603 ; http://n2t.net/addgene:63603 ; RRID:Addgene_63603) |
| 参考文献部分: | Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T. BMC Biotechnol. 2015 May 22;15(1):33. 10.1186/s12896-015-0144-x [pii] PubMed 25997509 |
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 pX330A_FokI-1x3 9476 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses FokI-dCas9 and gRNA.
ACCESSION .
VERSION .
KEYWORDS pX330A_FokI-1x3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9476)
AUTHORS Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T
TITLE Production of knockout mice by DNA microinjection of various
CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes.
JOURNAL BMC Biotechnol. 2015 May 22;15(1):33.
PUBMED 25997509
REFERENCE 2 (bases 1 to 9476)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9476
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 8..293
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 295..572
/label=chicken beta-actin promoter
intron 573..801
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 822..842
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 849..1436
/codon_start=1
/product="nonspecific DNA cleavage domain of the FokI
endonuclease (Li et al., 1992)"
/label=FokI cleavage domain
/note="must dimerize to be catalytically active"
/translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
CDS 1452..5552
/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 5553..5600
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5628..5645)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5634..5841
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5850..5990
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5850..5979
/label=AAV2 ITR
rep_origin 6065..6520
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6152..6171)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6362..6383
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6537..6556)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6656..6678
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6716..6734)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6802..6906
/gene="bla"
/label=AmpR promoter
CDS 6907..7767
/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(7125..7144)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7938..8526
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8427..8446
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 8588..8828
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 8588..8608
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 8759..8778
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 8855..8930
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS complement(9016..9321)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
primer_bind 9171..9193
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 9185..9202
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 9186..9202
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9212..9231
/label=T7
/note="T7 promoter, forward primer"
promoter 9212..9230
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9278..9298)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(9278..9296)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(9317..9333)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9317..9333)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9330..9352)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9341..9357
/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(9365..9395)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9410..9431
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
61 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
121 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
181 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
241 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
301 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
361 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg
421 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
481 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
541 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc
601 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
661 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
721 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
781 cctgaaatca ctttttttca ggttggaccg gtgccaccat gcctaagaag aagcggaagg
841 tgagctccca actcgtgaag agtgaacttg aggagaaaaa gtcggagctg cggcacaaat
901 tgaaatacgt accgcatgaa tacatcgaac ttatcgaaat tgctaggaac tcgactcaag
961 acagaatcct tgagatgaag gtaatggagt tctttatgaa ggtttatgga taccgaggga
1021 agcatctcgg tggatcacga aaacccgacg gagcaatcta tacggtgggg agcccgattg
1081 attacggagt gatcgtcgac acgaaagcct acagcggtgg gtacaatctt cccatcgggc
1141 aggcagatga gatgcaacgt tatgtcgaag aaaatcagac caggaacaaa cacatcaatc
1201 caaatgagtg gtggaaagtg tatccttcat cagtgaccga gtttaagttt ttgtttgtct
1261 ctgggcattt caaaggcaac tataaggccc agctcacacg gttgaatcac attacgaact
1321 gcaatggtgc ggttttgtcc gtagaggaac tgctcattgg tggagaaatg atcaaagcgg
1381 gaactctgac actggaagaa gtcagacgca agtttaacaa tggcgagatc aatttcggtg
1441 gcggtggatc cgacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct
1501 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
1561 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
1621 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
1681 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
1741 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
1801 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
1861 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
1921 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
1981 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
2041 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
2101 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
2161 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
2221 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
2281 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
2341 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
2401 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
2461 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
2521 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
2581 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
2641 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
2701 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
2761 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
2821 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
2881 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
2941 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
3001 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
3061 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
3121 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
3181 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
3241 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
3301 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
3361 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
3421 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
3481 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
3541 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
3601 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
3661 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
3721 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
3781 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
3841 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
3901 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
3961 tggacgccat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
4021 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
4081 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
4141 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
4201 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
4261 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
4321 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
4381 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
4441 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
4501 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
4561 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
4621 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
4681 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
4741 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
4801 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
4861 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
4921 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
4981 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
5041 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
5101 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
5161 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
5221 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
5281 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
5341 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
5401 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
5461 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
5521 cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg
5581 gccaggcaaa aaagaaaaag taagaattcc tagagctcgc tgatcagcct cgactgtgcc
5641 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
5701 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
5761 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
5821 gaatagcagg catgctgggg agcggccgca ggaaccccta gtgatggagt tggccactcc
5881 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
5941 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
6001 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
6061 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
6121 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
6181 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
6241 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
6301 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
6361 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
6421 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
6481 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
6541 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
6601 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
6661 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
6721 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
6781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
6841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
6901 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
6961 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
7021 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
7081 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
7141 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
7201 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
7261 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
7321 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
7381 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
7441 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
7501 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
7561 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
7621 ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
7681 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
7741 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
7801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
7861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
7921 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
7981 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
8041 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
8101 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
8161 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
8221 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
8281 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
8341 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
8401 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
8461 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
8521 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
8581 cacatgtgag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
8641 tagagagata attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
8701 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
8761 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
8821 gaaaggacga aacaccgggt cttcgagaag acctgtttta gagctagaaa tagcaagtta
8881 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt
8941 tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc
9001 tgcagacaaa tggctctaga gaccggcgcc gctacagggc gcgtcccatt cgccattcag
9061 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
9121 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
9181 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
9241 taccgggccc cccctcgagg tcctccagct tttgttccct ttagtgaggg ttaattgcgc
9301 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
9361 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
9421 aactcacatt aattgcgttg cgctcactgc ccgctttcca ccggtggtct ctctag
//
