| 质粒名称: | pcDNA3.1-hAsCpf1(TYCV) (pY210) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89351 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5402 bp |
| 完整质粒大小: | 9509 bp |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Acidaminococcus sp. Cpf1 (RR variant) |
| 物种来源: | Synthetic |
| 插入大小: | 4107 bp |
| 基因突变: | S542R/K607R |
| 启动子 | CMV |
| 标签/融合蛋白: |
NLS（插入件上的N端子）
NLS（插入件上的C端子）
3xHA（插入件上的C端子）
|
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | CGCAAATGGGCGGTAGGCGTG |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1-hAsCpf1(TYCV) (pY210) was a gift from Feng Zhang (Addgene plasmid # 89351 ; http://n2t.net/addgene:89351 ; RRID:Addgene_89351) |
| 参考文献部分: | Engineered Cpf1 variants with altered PAM specificities. Gao L, Cox DBT, Yan WX, Manteiga JC, Schneider MW, Yamano T, Nishimasu H, Nureki O, Crosetto N, Zhang F. Nat Biotechnol. 2017 Jun 5. doi: 10.1038/nbt.3900. 10.1038/nbt.3900 PubMed 28581492 |
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pcDNA3.1-hAsCpf1(TYCV) (pY210)载体质粒基本信息
pcDNA3.1-hAsCpf1(TYCV) (pY210)质粒图谱载体图谱和pcDNA3.1-hAsCpf1(TYCV) (pY210)载体序列质粒序列多克隆位点信息
 (pY210)质粒图谱.png)
pcDNA3.1-hAsCpf1(TYCV) (pY210)质粒载体简介
pcDNA3.1-hAsCpf1(TYCV) (pY210)质粒序列载体序列
LOCUS pcDNA3.1-hAsCpf1(TYCV) (pY210) 9509 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses humanized AsCpf1 variant that recognizes TYCV PAMs.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1-hAsCpf1(TYCV) (pY210)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9509)
AUTHORS Gao L, Cox DBT, Yan WX, Manteiga JC, Schneider MW, Yamano T,
Nishimasu H, Nureki O, Crosetto N, Zhang F
TITLE Engineered Cpf1 variants with altered PAM specificities.
JOURNAL Nat Biotechnol. 2017 Jun 5. doi: 10.1038/nbt.3900.
PUBMED 28581492
REFERENCE 2 (bases 1 to 9509)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9509
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 213..233
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(242..670)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(352..373)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 564..583
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 716..940
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 929..946
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
CDS complement(1020..1100)
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
CDS complement(1107..1154)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS complement(1155..5072)
/codon_start=1
/product="CRISPR-associated protein from Acidaminococcus
sp. BV3L6 (Zetsche et al., 2015)"
/label=AsCpf1
/translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
NYATKKPYSVEKFKLNFQMPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
LSFEPTEKTSEGFDKMYYDYFPDAAKMIPRCSTQLKAVTAHFQTHTTPILLSNNFIEPL
EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
DLKLQNGISNQDWLAYIQELRN"
regulatory 1426..1435
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS complement(5097..5117)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory 5120..5129
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(5164..5183)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5165..5183)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(5228..5431)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(5257..5277)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 5432..5811
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 5983..6002
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 6077..6181
/gene="bla"
/label=AmpR promoter
CDS 6182..7042
/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(6400..6419)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7213..7801
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7702..7721
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7955..7972
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 8089..8110
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8125..8155
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8163..8179
/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 8168..8190
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 8187..8203
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8187..8203
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 8252..8373
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8263..8282)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 8317..8336
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(8547..9341)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(8658..8677)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 9268..9287
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(9408..228)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 9422..48
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(9476..9495)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
ORIGIN
1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
661 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
721 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
781 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
841 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
901 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
961 taaacgggcc ctctagactc gagcggccgc cactgtgctg gatatctgca gaattcttag
1021 gcatagtcgg ggacatcata tgggtatgca taatcaggca cgtcgtaggg ataagcgtaa
1081 tctggaacat cgtatgggta ggatcccttt ttcttttttg cctggccggc ctttttcgtg
1141 gccgccggcc ttttgttgcg cagctcctgg atgtaggcca gccagtcctg attggagatg
1201 ccgttctgca gcttcagatc cttgctctcc ttcaggtgat tcagcagcag ctggcccttc
1261 agggcgatgt ggtaggcgcc attggcatcg gcgtccatgg gccactctgg gttctgaaac
1321 cgggagtcga agcacacgcc attcagatcg cgcacggggc tgttgatata gtcctcgcct
1381 gtggcggcat tggagttccg catctgcagc acgctgcgga tcagggccac catggtgtcg
1441 atggcgtgag aatcgtcatt ctccagcagc tttggcagga tgttggagcc atccctgaac
1501 acgatgccct tctcctccag cagggcgatc agctcgttgg caggatacag gtcccggtat
1561 ctgccggtga atctgtgatt ctcgatcact ggcacgattc tcttgccggc gatgaaaggg
1621 gtgcccttgg cgtcaaactg tgtctcgttc ttctcgaaca cgatatccca tgcaggcata
1681 aagccgggca ggcccctctg gaaggacaga tttctgttca tcttaaagtg caggatgaag
1741 tcgccggttt tcacgtcgta gtgcagaaag tcgaagccct ccaggaagtg cttgcggctc
1801 tcgtgattct tgatggtttt ccacacgaag gggtccacga agccggtcag gggatcgatc
1861 ttagatgtat atggggcagg cacgtaaaac aggaagccag actgggtgcc catcttggca
1921 aaggaggtga actggtctgt cagctggtat gggttcagca cgcctcccac tttctctgct
1981 ggatagtcct tcagcaccag gcaattcagc ttatcgatca gcatcttctc gaactgctgg
2041 tacacggcct tctcggcgat gccggtcctc ttgctcttaa agccgaaatt caggttctcc
2101 agcaccacca cggcctggta gtggatcatc aggtccacga tctcgtggat gacctggctc
2161 agatagccct gcttcagatc cttgattgtg cccaccacag accaggcctg ccttgctgcc
2221 accctctcct tctccctgtt gtccagcttc ttctggtaat caaactgctg gatggtgttc
2281 aggctccgct gctccaggat cttgccggtg gagtcgatca ctgtgatata gatcaggttt
2341 ctctcgcccc gatcgatgcc gatgataggt gtctcggggt gctccttcag gtaggcattc
2401 accctctggt tgaacttaga tggggaattg gcggcctgat agttcagtgt gataggcacg
2461 tggaaaaaga acttgtcgct ggtaaagcgc ctatccttga tgatctcgtg agacacctcc
2521 ttggtgatca cgttgggcag cagggccctg gcctcatcag acaggtcgtg ggacagtctg
2581 tgattcacat agtcgtacag ctcctggtac agggtgtcgg ggattggggt tttctgatcc
2641 ttcagcttct tgttcagcat cttctctccc agccggtgtg ccatcctctt catcctggac
2701 ttagggcggt agaacagctc ggcctggcca ttcagcttga tgcttgtctt ggccaggttc
2761 tctggagaaa acagaccggt ccaatacagt gtgtgcagat taggcttgcc gtggtggccc
2821 ttggcaaagt ccttgttata gatctggaac aggtacagct tgcctgtctc cacggcatcc
2881 atgatctcct tctcggcgat tctctggaag ctgatgtggt acagcagggg attcagctcg
2941 gcatagtact cgcccaggtc cttatactga gaggatggcc gcaggctaga cagatcgata
3001 gaggttgtct tggtatactt ggacagaaaa tcccttgtga agtcgatcca cttgcacagg
3061 gcctctctgt agcccttctg gtcgccggtt ttcttggcgt acgctgtctg aaacttcttt
3121 ggctccttct caggattgtt caggtcgtag atctcctttg tgatctccag aggctcgatg
3181 aaattgttgg acagcaggat gggggttgtg tgggtctgaa agtgggcggt caccgccttc
3241 agctgggtgc tgcaccgtgg gatcatcttg gcggcatccg gaaagtagtc atagtacatc
3301 ttatcaaagc cctcgctggt tttctctgtg ggctcgaagc tgagggcctt atacctgccc
3361 ttctgctttg gcatgatgcc cagatagtac aggccgttct tcacaaacag gatggcgcca
3421 ttgttcttct ccttattcac gtcccagccc cgggccagtg taggcatctg aaagttcagc
3481 ttgaacttct ccacggagta gggcttcttg gtggcataat ttctggcctt gttgtagaag
3541 ctcagagaag gctccatctc cagcttgatg ccggtcagcc gggcagagaa ctcggggtcc
3601 acctcgttgg actcatccac ggcaaaccag tccagcaggt ggtacaggcc cagcaggctg
3661 tccagctgag acttcaggat ctccttctcc tcctgcttct tcagggttgt aggcagtggc
3721 tgatccaggg cggcgtgtgc gtgggacagg atctcgctgg ttttctgctt gaaggcctcg
3781 ctcagctcct tgcctgcggc agagatgatc tcctgcaggt tgatatcctc gtgcttcagg
3841 ctgcgctgca ccttctcctt ggcagacttg gtgatcttgc ctgtcagctc ggagattctc
3901 cgctcataca gggcattcct cagtgtatcc cagtggtcgc acagggcgct gctgattgtc
3961 tccagcttct tgtggctgat gaagatgtgt gtcaggtcga tgctgttcag ctcgttaaac
4021 agggcctcgg ctgtctccag cacgttctcg tttctcagca gtgtcttgta cttgcagaag
4081 gactggatca cttcctcgtc gctcttaaac tcctccagga tgaaagacag ggtgttccta
4141 tcggacagga tctgcttaaa cagggggatg aatctgtgtg gcagggaggc gatgatgtgg
4201 gctgtctcat cattcttctg gatggccaga ttcagcacct cgttcaggcc cttgatcttc
4261 tcggtgcctg cctcccgaga gattcctccc agcagctggt tatacaggtc gatctgggtc
4321 tgtgtcagca gctggttata aaaagggaag gaaaacacct cctcgatgga ggtgctcacg
4381 aagatgccga tggccttctt cacgttctca aagtgctccc gcaggctggg cacggcggtg
4441 atcaggcgtg tgaagatgtg acaattctcc ttaaacttgg ggaagttgtc ctgcacgatg
4501 cggtgtggga tggctgtgct gatatcctcg gcgctgaaca cgttcttcct gttctcataa
4561 aagccggaga agtaggttgt aaacttgtcg aagctccgca gcagggcgtt ctcgtgctcg
4621 gttgtggtca cggtgcccag ctgcttcagc accttgccat taaacagctc ggccttgaac
4681 aggcccttgt agatctcggc gtgtctctta ttgatggcat cggtcaggtt gtctgtccgg
4741 ccgatgaagt agtcgtggat ggcattgcga tatgtggcct gctcctcgat cagggcgttc
4801 cttgtctcct cggttttctc ctttctatag gagtcgatgg cggcgctcag gttctcccaa
4861 tccagctgca ccagctgcag gcactggtcg gcataggtct tgtagatccg atcgatgatg
4921 ggcttcagct ccttgtagtg atcattgcgg gccttgtcct cctcgatgaa gccctgctcc
4981 tggatgtgct tcagggtctt gccctgtggg atcagctcaa accgcagtgt cttgctcacc
5041 tgatacaggt tggtaaagcc ctcgaactgt gtggctgctg ggactccgtg gataccgacc
5101 ttccgcttct tctttggggc catggtggca agcttaagtt taaacgctag ccagcttggg
5161 tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta
5221 gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
5281 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
5341 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
5401 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
5461 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
5521 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
5581 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
5641 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
5701 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
5761 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
5821 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
5881 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
5941 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
6001 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
6061 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
6121 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
6181 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
6241 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
6301 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
6361 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
6421 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
6481 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
6541 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
6601 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
6661 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
6721 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
6781 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
6841 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
6901 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
6961 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
7021 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
7081 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
7141 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
7201 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
7261 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
7321 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
7381 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
7441 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
7501 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
7561 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
7621 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
7681 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
7741 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
7801 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
7861 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
7921 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
7981 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
8041 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
8101 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
8161 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
8221 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc
8281 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
8341 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
8401 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
8461 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
8521 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
8581 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
8641 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
8701 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
8761 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
8821 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
8881 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
8941 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
9001 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
9061 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
9121 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
9181 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
9241 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
9301 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
9361 gtctcttgat cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct
9421 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg
9481 gcctctgcat aaataaaaaa aattagtca
//
