| 质粒名称: | PCS2+Cas9-mSA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 103882 |
| 存储实验室: | anet Rossant |
|
载体骨架基本信息 |
|
| 载体名称: | JPCS2+ |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4072 bp |
| 完整质粒大小: | 8886 bp |
| 载体类型: | CRISPR |
| 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: | CAS9-MSA |
| 物种来源: | synthetic |
| 插入大小: | 4818 bp |
| 启动子 | SP6 |
| 标签/融合蛋白: | MSA（单体链霉亲和素（插入时的C端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Kpn1(未破坏) |
| 3' 克隆位点: | 已销毁（如果已销毁则未知） |
| 5' 测序引物 | ATTTAGGTGACACTAT |
| 3' 测序引物 | CCTGTGTGAAATTGTTATCC |
|
文献引用方法 |
|
| 材料和方法部分: | PCS2+Cas9-mSA was a gift from Janet Rossant (Addgene plasmid # 103882 ; http://n2t.net/addgene:103882 ; RRID:Addgene_103882) |
| 参考文献部分: | Efficient generation of targeted large insertions by microinjection into two-cell-stage mouse embryos. Gu B, Posfai E, Rossant J. Nat Biotechnol. 2018 Jun 11. pii: nbt.4166. doi: 10.1038/nbt.4166. 10.1038/nbt.4166 PubMed 29889212 |
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 PCS2+Cas9-mSA 8883 bp ds-DNA circular SYN 07-NOV-2018
DEFINITION Cas9 mSA for in vitro transcription for gene editing in Mouse
Embryos.
ACCESSION .
VERSION .
KEYWORDS PCS2+Cas9-mSA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8883)
AUTHORS Gu B, Posfai E, Rossant J
TITLE Efficient generation of targeted large insertions by microinjection
into two-cell-stage mouse embryos.
JOURNAL Nat Biotechnol. 2018 Jun 11. pii: nbt.4166. doi: 10.1038/nbt.4166.
PUBMED 29889212
REFERENCE 2 (bases 1 to 8883)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 7, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8883
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..978
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1013..1031
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1013..1030
/label=SP6
/note="SP6 promoter, forward primer"
CDS 1066..1131
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1138..1158
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1183..5283
/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 5284..5331
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5401..5463
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
/translation="METDTLLLWVLLLWVPGSTGD"
CDS 5464..5490
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(5891..5908)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 5913..6047
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5937..5956)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5991..6010
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(6160..6180)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(6160..6178)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6199..6215)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6199..6215)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6212..6234)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6223..6239
/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(6247..6277)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6292..6313
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6430..6447)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6601..7189)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6681..6700)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7360..8220)
/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 7983..8002
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8221..8325)
/gene="bla"
/label=AmpR promoter
rep_origin complement(8351..8806)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8488..8509)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 8700..8719
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
1021 gacactatag aatacaagct acttgttctt tttgcaggat ccatggacta taaggaccac
1081 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca
1141 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc
1201 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
1261 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
1321 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
1381 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
1441 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
1501 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
1561 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
1621 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
1681 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
1741 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
1801 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
1861 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
1921 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
1981 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
2041 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
2101 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
2161 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
2221 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
2281 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
2341 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
2401 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
2461 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
2521 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
2581 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
2641 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
2701 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
2761 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
2821 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
2881 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
2941 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
3001 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
3061 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
3121 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
3181 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
3241 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
3301 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
3361 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
3421 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
3481 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
3541 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
3601 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
3661 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
3721 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
3781 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
3841 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
3901 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
3961 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
4021 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
4081 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
4141 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
4201 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
4261 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
4321 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
4381 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
4441 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
4501 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
4561 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
4621 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
4681 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
4741 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
4801 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
4861 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
4921 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
4981 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
5041 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
5101 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
5161 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
5221 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
5281 gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gtatgaattc
5341 aaggcctctc gaagcggttc agagacccca ggaactagcg agagcgctac accggaatcg
5401 atggagacag acacactcct gctatgggta ctgctgctct gggttccagg ttccactggt
5461 gactatccat atgatgttcc agattatgct ggggcccagc cggccagatc tatggcggaa
5521 gcgggtatca ccggcacgtg gtacaaccag tctggttcta ccttcaccgt taccgcgggt
5581 gcggacggta acctgaccgg tcagtacgaa aaccgtgcgc agggcactgg ttgccagaac
5641 tctccgtaca ccctgaccgg tcgttacaac ggtaccaaac tggaatggcg tgttgaatgg
5701 aacaactcta ccgaaaactg ccactctcgt accgaatggc gtggtcagta ccagggtggt
5761 gcggaagcgc gtatcaacac ccagtggaac ctgacctacg aaggtggttc tggtccggcg
5821 accgaacagg gtcaggacac cttcaccaaa gttaaaccgt ctgcggcgtc tggatcctga
5881 gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
5941 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
6001 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
6061 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
6121 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
6181 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
6241 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
6301 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
6361 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
6421 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
6481 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
6541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
6601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
6661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
6721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
6781 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
6841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
6901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
6961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
7021 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
7081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
7141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
7201 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
7261 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
7321 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
7381 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
7441 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
7501 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
7561 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
7621 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
7681 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
7741 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
7801 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
7861 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
7921 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
7981 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
8041 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
8101 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
8161 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
8221 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
8281 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
8341 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
8401 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
8461 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
8521 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
8581 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
8641 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
8701 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
8761 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
8821 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
8881 acc
//
