| 质粒名称: | pCS2-nCas9n-nanos 3’UTR |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 62542 |
| 存储实验室: | Antonio Giraldez |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2-nCas9n |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 8272 bp |
| 完整质粒大小: | 8910 bp |
| 载体修饰: | -- |
| 载体类型: | 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: | nanos 3' UTR,nos1 |
| 物种来源: | D. rerio (zebrafish) |
| 插入大小: | -- |
| 基因突变: | 3' UTR only |
| GenBank编号 | -- |
| 启动子 | SP6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SnaBI (在克隆过程中销毁) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | ATCaccggtAGCGGACATTGATGCTCC |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | pCS2-nCas9n-nanos 3’UTR was a gift from Antonio Giraldez (Addgene plasmid # 62542 ; http://n2t.net/addgene:62542 ; RRID:Addgene_62542) |
| 参考文献部分: | CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo. Moreno-Mateos MA, Vejnar CE, Beaudoin JD, Fernandez JP, Mis EK, Khokha MK, Giraldez AJ. Nat Methods. 2015 Aug 31. doi: 10.1038/nmeth.3543. 10.1038/nmeth.3543 PubMed 26322839 |
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-nCas9n-nanos 3'UTR 8903 bp ds-DNA circular SYN 15-FEB-2019
DEFINITION expression of a germ-line targeted, codon optimized Cas9 for
genome-editing in zebrafish.
ACCESSION .
VERSION .
KEYWORDS pCS2-nCas9n-nanos 3'UTR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8903)
AUTHORS Moreno-Mateos MA, Vejnar CE, Beaudoin JD, Fernandez JP, Mis EK,
Khokha MK, Giraldez AJ
TITLE CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9
targeting in vivo.
JOURNAL Nat Methods. 2015 Aug 31. doi: 10.1038/nmeth.3543.
PUBMED 26322839
REFERENCE 2 (bases 1 to 8903)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 15, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8903
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 35..52
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 99..108
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 117..137
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 144..4244
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes MGAS15252 Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL
ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
RBS 4157..4165
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 4260..4280
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
promoter complement(4299..4316)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(4933..4950)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 4955..5089
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4979..4998)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5033..5052
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(5202..5222)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5202..5220)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5241..5257)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5241..5257)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5254..5276)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5265..5281
/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(5289..5319)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5334..5355
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5472..5489)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5643..6231)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5723..5742)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6402..7262)
/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 7025..7044
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7263..7367)
/gene="bla"
/label=AmpR promoter
rep_origin complement(7393..7848)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7530..7551)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7742..7761
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 7926..8903
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
61 ctacttgttc tttttgcagg atccgatcca atgtcgacgc caccatggct tctccaccta
121 agaagaagag aaaggtggga agcatggata agaagtatag catcggcctg gatattggaa
181 ctaactccgt gggttgggca gtgattacag acgactacaa ggtccctagc aagaaattta
241 aggtgctggg taacaccgac aggcacagca tcaagaaaaa tctgattgga gccctgctgt
301 tcggttctgg agagactgcc gaagcaacac gcctgaaaag aacagcaaga aggcgctata
361 ccagaaggaa gaatagaatc tgttacctgc aggagatttt ctctaacgaa atggctaagg
421 tggacgattc attctttcat aggctggagg aaagtttcct ggtcgaggaa gataagaaac
481 acgagcgcca tcctatcttt ggaaacattg tggacgaggt cgcctatcac gaaaaatacc
541 caaccatcta tcatctgcgc aagaaactgg ctgactctac tgataaagcc gacctgagac
601 tgatctatct ggctctggcc cacatgatta agttcagggg tcattttctg atcgagggcg
661 atctgaaccc cgacaattcc gatgtggaca agctgttcat ccagctggtc cagatttaca
721 atcagctgtt tgaggaaaac cctattaatg cttccagagt ggacgcaaaa gctatcctgt
781 cagccaggct gtccaagtca cgcagactgg agaacctgat tgcacagctg cccggagaaa
841 agaggaacgg tctgtttgga aatctgatcg ctctgagtct gggcctgact cctaacttca
901 aaagcaattt tgatctggct gaggacgcca aactgcagct gtcaaaggac acatatgacg
961 atgacctgga taacctgctg gcacagatcg gagatcagta cgctgacctg ttcctggctg
1021 ccaaaaatct gtccgacgca atcctgctgt cagatattct gagagtgaac agcgagatta
1081 caaaagctcc tctgagtgcc agcatgatca agagatatga cgagcaccat caggatctga
1141 ccctgctgaa ggctctggtc aggcagcagc tgccagagaa gtacaaggaa attttctttg
1201 atcagtccaa gaacggctac gccggttata tcgacggagg cgcatcacag gaggaattct
1261 acaagtttat caaacctatt ctggagaaga tggacggaac tgaggaactg ctggtgaaac
1321 tgaatagaga ggacctgctg aggaagcagc gcacatttga taacggttcc atcccacacc
1381 agattcatct gggagagctg cacgctatcc tgaggcgcca ggaagacttc tacccctttc
1441 tgaaagataa ccgcgagaag atcgaaaaaa ttctgacctt cagaatccct tactatgtgg
1501 gtccactggc tcgcggaaac agcagatttg cctggatgac tcgcaaatcc gaggaaacca
1561 ttactccttg gaacttcgag gaagtggtcg ataagggcgc ctctgcacag tccttcatcg
1621 agagaatgac taattttgac aaaaacctgc ccaatgagaa agtgctgcct aagcactccc
1681 tgctgtacga gtatttcact gtctataacg aactgacaaa ggtgaaatac gtcaccgagg
1741 gcatgagaaa gccagccttc ctgtcaggag agcagaagaa agcaatcgtg gatctgctgt
1801 ttaaaaccaa caggaaagtg actgtcaagc agctgaagga ggactacttc aagaaaattg
1861 aatgcttcga ttccgtggag atcagcggag tcgaagacag atttaacgca agcctgggcg
1921 cttaccacga tctgctgaag atcattaagg ataaagactt cctggacaac gaggaaaatg
1981 aggatatcct ggaagacatt gtgctgacac tgaccctgtt tgaggacaga ggaatgatcg
2041 aggaaagact gaaaacctat gctcatctgt tcgatgacaa ggtgatgaaa cagctgaaga
2101 gaaggcgcta cactggctgg ggtagactga gcaggaagct gatcaacggc attagggata
2161 aacagtcagg aaagacaatc ctggactttc tgaaaagtga tggcttcgcc aaccgcaatt
2221 ttatgcagct gattcacgat gacagtctga ccttcaaaga ggacatccag aaggctcagg
2281 tgtctggaca gggccactcc ctgcatgagc agattgcaaa cctggctgga agcccagcca
2341 tcaagaaagg cattctgcag acagtgaaaa tcgtcgatga gctggtgaaa gtcatgggcc
2401 ataagcccga aaacatcgtg attgagatgg ctcgcgaaaa tcagacaacc cagaagggtc
2461 agaagaacag tagagagagg atgaaaagaa tcgaggaagg cattaaggag ctgggtagcc
2521 agatcctgaa agagcaccca gtggaaaaca cacagctgca gaatgagaag ctgtatctgt
2581 actatctgca gaatggaaga gatatgtacg tggaccagga gctggatatt aacaggctgt
2641 ctgattacga cgtggatcat atcgtccccc agagtttcat caaagatgac agcattgaca
2701 acaaggtgct gaccaggtcc gacaaaaaca gaggaaaatc agataatgtc cctagtgagg
2761 aagtggtcaa gaaaatgaag aactactgga gacagctgct gaatgccaaa ctgatcactc
2821 agaggaagtt tgataacctg acaaaagcag agcgcggtgg actgtcagaa ctggacaaag
2881 ctggattcat caagaggcag ctggtggaaa cacgccagat cactaaacac gtcgcacaga
2941 ttctggatag tcgcatgaac acaaagtacg atgagaatga caaactgatc agagaagtga
3001 aggtcattac cctgaagagt aaactggtca gcgactttag gaaagatttc cagttttata
3061 aggtccgcga gattaacaat tatcaccatg cccatgacgc atacctgaac gccgtggtcg
3121 gtaccgcact gatcaagaaa tacccaaaac tggagagcga attcgtgtac ggagactata
3181 aggtgtacga tgtcagaaaa atgatcgcca agtccgagca ggaaattgga aaagctactg
3241 ccaagtattt cttttactca aacatcatga atttctttaa gacagagatc accctggcca
3301 atggagaaat ccgcaaaagg cccctgattg agacaaacgg agagacaggc gaaatcgtgt
3361 gggacaaagg cagagatttt gcaaccgtga ggaaggtcct gagcatgcct caagtgaata
3421 tcgtcaagaa aactgaggtg cagacaggcg gtttctcaaa agaaagtatt ctgccaaaac
3481 gcaactctga taagctgatc gctagaaaga aagactggga ccctaagaag tatggaggct
3541 ttgactctcc cactgtggca tactccgtcc tggtggtcgc taaggtggag aagggcaaaa
3601 gcaagaaact gaaatctgtc aaggagctgc tgggtatcac aattatggag agaagctctt
3661 tcgagaagaa cccaatcgat tttctggagg ccaaaggtta taaggaagtg aagaaagacc
3721 tgatcattaa actgcccaag tacagtctgt ttgagctgga aaacggcagg aaacgcatgc
3781 tggcaagcgc tggagagctg cagaaaggca atgaactggc cctgccttct aagtacgtga
3841 acttcctgta tctggcaagc cactacgaga agctgaaagg atctccagag gataacgaac
3901 agaaacagct gtttgtggag cagcacaagc attatctgga cgagatcatt gaacagatta
3961 gcgagttctc taaaagagtg atcctggccg acgcaaatct ggataaggtc ctgtctgctt
4021 acaacaaaca cagagataag cccatcaggg agcaggccga aaatatcatt catctgttca
4081 ctctgacaaa cctgggcgca cctgcagctt tcaagtactt cgacactaca atcgatagaa
4141 agaggtacac ctccactaag gaggtgctgg acgctacact gatccatcag agtattaccg
4201 gcctgtacga aacaaggatt gacctgtctc agctgggtgg cgactcaccg gtgagatctc
4261 ctaagaagaa gagaaaggtg tgaccgcgga atctagaact atagtgagtc gtattactca
4321 ccggtagcgg acattgatgc tccgggagat ttgaagaaac actttttacc gcaggtttta
4381 atgtttaagt tttaactctt taattgtttg tttggttgat acgcggcgga ttgcgagttt
4441 gcatgcatgt gtgcgttcac tgtttgattt tgcacttttt ttgtgtgtgt gtatatgtgt
4501 gtgtttgctg tgttttattt tgtgtgcact ggtgttgtgt tttcacttgg taacaaactc
4561 gtacacaagc cagcaggctc gctacaggcg caaccgcact caaaaacaaa ccctttcatg
4621 cttatttggt aaatacaatg tgtgtttagt cctcctttta aatgtcagat tttatggtgt
4681 tgtatttaaa caaaaaattc aatgttaata tttagatttt agtgatttta ttattgaaaa
4741 cggcttgttt tgtataagta acctttaaaa aaagttttct ccattgcatt taaattcagc
4801 ttgaaaaaca taatcgccat attttcatgt cgcttgctaa aattcatgta ctactttcat
4861 cattttatgt cagtgtgtga tttttgactt gtgatggagt gaaaaatgtg aggaaaatat
4921 aaacatctag aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga
4981 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
5041 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
5101 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc
5161 cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt
5221 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
5281 attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg
5341 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
5401 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
5461 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
5521 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
5581 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
5641 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
5701 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
5761 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
5821 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
5881 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
5941 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
6001 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
6061 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
6121 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
6181 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
6241 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
6301 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
6361 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
6421 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
6481 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
6541 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
6601 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
6661 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
6721 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
6781 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
6841 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
6901 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
6961 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
7021 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
7081 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
7141 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
7201 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
7261 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
7321 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
7381 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
7441 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
7501 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
7561 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
7621 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
7681 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
7741 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
7801 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
7861 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt
7921 cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga
7981 cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga
8041 ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc
8101 actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg
8161 ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc
8221 tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
8281 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
8341 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
8401 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
8461 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
8521 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
8581 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
8641 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
8701 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
8761 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
8821 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
8881 taagcaatgc tcgtttaggg aac
//
