| 质粒名称: | 6His-MBP-TEV-huAsCpf1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 90095 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pET-28 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6643 bp |
| 完整质粒大小: | 10710 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | huAsCpf1 |
| 物种来源: | humanized Acidaminococcus sp. BV3L6 Cpf1 |
| 插入大小: | 4056 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
6xHis（主干上的N终端）
MBP（插入件上的N端子）
TEV site（插入件上的N端子）
Nucleoplasmin NLS（插入物上的C末端）
3xHA（插入件上的C端子）
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | 6His-MBP-TEV-huAsCpf1 was a gift from Feng Zhang (Addgene plasmid # 90095 ; http://n2t.net/addgene:90095 ; RRID:Addgene_90095) |
| 参考文献部分: | Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS, Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F. Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5. 10.1038/nbt.3737 PubMed 27918548 |
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 6His-MBP-TEV-huAsCpf1 10706 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacterial expression plasmid for protein purification.
ACCESSION .
VERSION .
KEYWORDS 6His-MBP-TEV-huAsCpf1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10706)
AUTHORS Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J,
DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS,
Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F
TITLE Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array.
JOURNAL Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub
2016 Dec 5.
PUBMED 27918548
REFERENCE 2 (bases 1 to 10706)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10706
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1334..1525
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(1519..1541)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 1627..1766
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 1682..1701
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(1781..1798)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1952..2540)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2032..2051)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2711..3571)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3334..3353
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3572..3676)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3702..4157)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3839..3860)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4051..4070
/label=F1ori-R
/note="F1 origin, reverse primer"
terminator 4194..4241
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 4237..4255
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(4308..4325)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(4341..4385)
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS complement(4386..4409)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS complement(4434..4514)
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
CDS complement(4521..4568)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS complement(4569..8489)
/codon_start=1
/product="CRISPR-associated protein from Acidaminococcus
sp. BV3L6 (Zetsche et al., 2015)"
/label=AsCpf1
/translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
KDLKLQNGISNQDWLAYIQELRN"
regulatory 4840..4849
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS complement(8490..8510)
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS complement(8517..8534)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
regulatory 8687..8696
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS complement(9651..9668)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 9682..9704
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 9719..9743
/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 complement(9743..9762)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(9744..9762)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9846..9865)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 10071..10148
/gene="lacI"
/label=lacI promoter
CDS 10149..525
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(10168..10187)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
61 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
121 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
181 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
241 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
301 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
361 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
421 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
541 tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa
601 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
661 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
721 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
781 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
841 ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag
901 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
961 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
1021 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
1081 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
1141 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
1201 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
1261 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
1321 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
1381 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
1441 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
1501 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
1561 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
1621 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
1681 gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat
1741 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
1801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
1861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
1921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
1981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
2041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
2101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
2161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
2221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
2281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
2341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
2401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
2461 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
2521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
2581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
2641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
2701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
2761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
2821 catctggccc cagtgctgca atgataccgc gactaccacg ctcaccggct ccagatttat
2881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
2941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
3001 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
3061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
3121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
3181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
3241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
3301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
3361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
3421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
3481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
3541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
3601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
3661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
3721 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
3781 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
3841 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
3901 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
3961 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
4021 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
4081 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
4141 gccgctacag ggcgcgtccc attcgccaat ccggatatag ttcctccttt cagcaaaaaa
4201 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca
4261 gcagccaact cagcttcctt tcgggctttg ttagcagccg gatctcagtg gtggtggtgg
4321 tggtgctcga caagctttta ctcgtgccat tcaattttct gggcttcaaa gatatcgttc
4381 aggcctttat catcatcatc tttataatcg ctaccaccac cctcgaggct ttaggcatag
4441 tcggggacat catatgggta tgcataatca ggcacgtcgt agggataagc gtaatctgga
4501 acatcgtatg ggtaggatcc ctttttcttt tttgcctggc cggccttttt cgtggccgcc
4561 ggccttttgt tgcgcagctc ctggatgtag gccagccagt cctgattgga gatgccgttc
4621 tgcagcttca gatccttgct ctccttcagg tgattcagca gcagctggcc cttcagggcg
4681 atgtggtagg cgccattggc atcggcgtcc atgggccact ctgggttctg aaaccgggag
4741 tcgaagcaca cgccattcag atcgcgcacg gggctgttga tatagtcctc gcctgtggcg
4801 gcattggagt tccgcatctg cagcacgctg cggatcaggg ccaccatggt gtcgatggcg
4861 tgagaatcgt cattctccag cagctttggc aggatgttgg agccatccct gaacacgatg
4921 cccttctcct ccagcagggc gatcagctcg ttggcaggat acaggtcccg gtatctgccg
4981 gtgaatctgt gattctcgat cactggcacg attctcttgc cggcgatgaa aggggtgccc
5041 ttggcgtcaa actgtgtctc gttcttctcg aacacgatat cccatgcagg cataaagccg
5101 ggcaggcccc tctggaagga cagatttctg ttcatcttaa agtgcaggat gaagtcgccg
5161 gttttcacgt cgtagtgcag aaagtcgaag ccctccagga agtgcttgcg gctctcgtga
5221 ttcttgatgg ttttccacac gaaggggtcc acgaagccgg tcaggggatc gatcttagat
5281 gtatatgggg caggcacgta aaacaggaag ccagactggg tgcccatctt ggcaaaggag
5341 gtgaactggt ctgtcagctg gtatgggttc agcacgcctc ccactttctc tgctggatag
5401 tccttcagca ccaggcaatt cagcttatcg atcagcatct tctcgaactg ctggtacacg
5461 gccttctcgg cgatgccggt cctcttgctc ttaaagccga aattcaggtt ctccagcacc
5521 accacggcct ggtagtggat catcaggtcc acgatctcgt ggatgacctg gctcagatag
5581 ccctgcttca gatccttgat tgtgcccacc acagaccagg cctgccttgc tgccaccctc
5641 tccttctccc tgttgtccag cttcttctgg taatcaaact gctggatggt gttcaggctc
5701 cgctgctcca ggatcttgcc ggtggagtcg atcactgtga tatagatcag gtttctctcg
5761 ccccgatcga tgccgatgat aggtgtctcg gggtgctcct tcaggtaggc attcaccctc
5821 tggttgaact tagatgggga attggcggcc tgatagttca gtgtgatagg cacgtggaaa
5881 aagaacttgt cgctggtaaa gcgcctatcc ttgatgatct cgtgagacac ctccttggtg
5941 atcacgttgg gcagcagggc cctggcctca tcagacaggt cgtgggacag tctgtgattc
6001 acatagtcgt acagctcctg gtacagggtg tcggggattg gggttttctg atccttcagc
6061 ttcttgttca gcatcttctc tcccagccgg tgtgccatcc tcttcatcct ggacttaggg
6121 cggtagaaca gctcggcctg gccattcagc ttgatgcttg tcttggccag gttctctgga
6181 gaaaacaggc cggtccaata cagtgtgtgc agattaggct tgccgtggtg gcccttggca
6241 aagtccttgt tatagatctg gaacaggtac agcttgcctg tctccacggc atccatgatc
6301 tccttctcgg cgattctctg gaagctgatg tggtacagca ggggattcag ctcggcatag
6361 tactcgccca ggtccttata ctgagaggat ggccgcaggc tagacagatc gatagaggtt
6421 gtcttggtat acttggacag aaaatccctt gtgaagtcga tccacttgca cagggcctct
6481 ctgtagccct tctggtcgcc ggttttcttg gcgtaggctg tctgaaactt ctttggctcc
6541 ttctcaggat tgttcaggtc gtagatctcc tttgtgatct ccagaggctc gatgaaattg
6601 ttggacagca ggatgggggt tgtgtgggtc tgaaagtggg ctgtcacggc cttcagctgg
6661 gtgctgcact ttgggatcat cttggcggca tcagggaagt agtcatagta catcttatca
6721 aagccctcgc tggttttctc tgtgggctcg aagctcaggg ccttatacct gcccttctgc
6781 tttggcatga tgcccagata gtacaggccg ttcttcacaa acaggatggc gccattgttc
6841 ttctccttat tcacgtccca gccagaggcc agtgtaggca tctgaaagtt cagcttgaac
6901 ttctccacgg agtagggctt cttggtggca taatttctgg ccttgttgta gaagctcaga
6961 gaaggctcca tctccagctt gatgccggtc agccgggcag agaactcggg gtccacctcg
7021 ttggactcat ccacggcaaa ccagtccagc aggtggtaca ggcccagcag gctgtccagc
7081 tgagacttca ggatctcctt ctcctcctgc ttcttcaggg ttgtaggcag tggctgatcc
7141 agggcggcgt gtgcgtggga caggatctcg ctggttttct gcttgaaggc ctcgctcagc
7201 tccttgcctg cggcagagat gatctcctgc aggttgatat cctcgtgctt caggctgcgc
7261 tgcaccttct ccttggcaga cttggtgatc ttgcctgtca gctcggagat tctccgctca
7321 tacagggcat tcctcagtgt atcccagtgg tcgcacaggg cgctgctgat tgtctccagc
7381 ttcttgtggc tgatgaagat gtgtgtcagg tcgatgctgt tcagctcgtt aaacagggcc
7441 tcggctgtct ccagcacgtt ctcgtttctc agcagtgtct tgtacttgca gaaggactgg
7501 atcacttcct cgtcgctctt aaactcctcc aggatgaaag acagggtgtt cctatcggac
7561 aggatctgct taaacagggg gatgaatctg tgtggcaggg aggcgatgat gtgggctgtc
7621 tcatcattct tctggatggc cagattcagc acctcgttca ggcccttgat cttctcggtg
7681 cctgcctccc gagagattcc tcccagcagc tggttataca ggtcgatctg ggtctgtgtc
7741 agcagctggt tataaaaagg gaaggaaaac acctcctcga tggaggtgct cacgaagatg
7801 ccgatggcct tcttcacgtt ctcaaagtgc tcccgcaggc tgggcacggc ggtgatcagg
7861 cgtgtgaaga tgtgacaatt ctccttaaac ttggggaagt tgtcctgcac gatgcggtgt
7921 gggatggctg tgctgatatc ctcggcgctg aacacgttct tcctgttctc ataaaagccg
7981 gagaagtagg ttgtaaactt gtcgaagctc cgcagcaggg cgttctcgtg ctcggttgtg
8041 gtcacggtgc ccagctgctt cagcaccttg ccattaaaca gctcggcctt gaacaggccc
8101 ttgtagatct cggcgtgtct cttattgatg gcatcggtca ggttgtctgt ccggccgatg
8161 aagtagtcgt ggatggcatt gcgatatgtg gcctgctcct cgatcagggc gttccttgtc
8221 tcctcggttt tctcctttct ataggagtcg atggcggcgc tcaggttctc ccaatccagc
8281 tgcaccagct gcaggcactg gtcggcatag gtcttgtaga tccgatcgat gatgggcttc
8341 agctccttgt agtgatcatt gcgggccttg tcctcctcga tgaagccctg ctcctggatg
8401 tgcttcaggg tcttgccctg tgggatcagc tcaaaccgca gtgtcttgct cacctgatac
8461 aggttggtaa agccctcgaa ctgtgtcatt ccctgaaaat acagattttc gctagcgcta
8521 ccacgcggaa ccagggatcc ggtgcttgca ccggtctggg catctttcag ggcttcatca
8581 acggtctgac gacctgatgc ggcattaatc actgcggtac gaacggcata ccaaaatgcg
8641 ctcatctgcg gaatattcgg cataatttcg cctttctgtg cattttccat ggtggctgca
8701 atacgcggat ctttaaccag ttcttcttca tagcttttca gtgcaactgc acccagcggt
8761 ttatctttat tcactgcttc cagaccttca tcggtcagca gatagttttc cagaaattct
8821 tttgccagtt ctttgttcgg gcttgctgca ttaatacctg cgctcagaac accaacaaac
8881 ggtttgctcg gctgaccttt aaatgtcggc agaacggtaa caccataatt cactttgctg
8941 gtatcaatat ttgaccatgc ccacggacca ttaattgtca ttgcggtttc acctttgtta
9001 aatgctgctt ctgcaatgct ataatcggta tcggcgttca tgtgtttgtt tttaatcaga
9061 tcaaccagaa aggtcagacc ggcttttgca cccgcattat caacacccac gtctttgata
9121 tcatatttgc cgttttcata tttgaatgca tagccaccat ctgctgcaat cagaggccag
9181 gtaaaatacg gttcttgcag attaaacatc agtgcgcttt taccttttgc tttcagttct
9241 ttatccagtg ccggaatttc ttcccaggtt ttaggcggat tcggcagcag atctttgtta
9301 tagatcaggc tcagtgcttc aactgcaatc ggatatgcaa tcagtttacc gttatagcga
9361 actgcatccc aggtaaacgg atacagtttg tcctgaaatg ctttatccgg tgtaatttct
9421 gccagcagac cgctctgtgc ataaccacca aaacgatcat gtgcccaaaa gataatatcc
9481 ggaccatcac cggttgctgc aacctgcgga aatttttctt ccagtttatc cggatgttca
9541 acggtcactt taatgccggt atctttttcg aattttttgc caacttctgc cagaccatta
9601 tagcctttat cgccattgat ccaaataacc agtttgcctt cttcgatttt gtgatggtga
9661 tgatgatgac ccatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
9721 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
9781 ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
9841 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
9901 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
9961 catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
10021 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg
10081 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
10141 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
10201 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
10261 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
10321 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
10381 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
10441 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
10501 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
10561 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
10621 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
10681 gtcaccagca aatcgcgctg ttagcg
//
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