| 质粒名称: | 6His-MBP-TEV-huLbCpf1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 90096 |
| 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: | 3819 bp |
| 完整质粒大小: | 10473 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 宿主菌: | BL21(DE3)pLysS |
| 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: | huLbCpf1 |
| 物种来源: | humanized Lachnospiraceae bacterium ND2006 Cpf1 |
| 插入大小: | 3819 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-huLbCpf1 was a gift from Feng Zhang (Addgene plasmid # 90096 ; http://n2t.net/addgene:90096 ; RRID:Addgene_90096) |
| 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-huLbCpf1 10469 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacterial expression plasmid for protein purification.
ACCESSION .
VERSION .
KEYWORDS 6His-MBP-TEV-huLbCpf1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10469)
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 10469)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10469
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 939..958
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(978..1055)
/gene="lacI"
/label=lacI promoter
primer_bind 1261..1280
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 1364..1383
/label=T7
/note="T7 promoter, forward primer"
promoter 1364..1382
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1383..1407
/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)."
RBS 1422..1444
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1458..1475
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
regulatory 2430..2439
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2592..2609
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 2616..2636
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 2637..6320
/codon_start=1
/product="CRISPR-associated protein from Lachnospiraceae
bacterium ND2006 (Zetsche et al., 2015)"
/label=LbCpf1
/translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
VKH"
CDS 6321..6368
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6375..6455
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
CDS 6480..6503
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 6504..6548
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 6564..6581
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6634..6652)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6648..6695
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 6732..7187
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6819..6838)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7029..7050
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7213..7317
/gene="bla"
/label=AmpR promoter
CDS 7318..8178
/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 complement(7536..7555)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8349..8937
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8838..8857
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9091..9108
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 9123..9262
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(9188..9207)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9348..9370
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(9364..9555)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(10364..977)
/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"
ORIGIN
1 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
61 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
121 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
181 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
241 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
301 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
361 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
421 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
481 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
541 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
601 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
661 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
721 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
781 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
841 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
901 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
961 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
1021 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
1081 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
1141 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
1201 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
1261 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
1321 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
1381 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
1441 gagatataca tatgggtcat catcatcacc atcacaaaat cgaagaaggc aaactggtta
1501 tttggatcaa tggcgataaa ggctataatg gtctggcaga agttggcaaa aaattcgaaa
1561 aagataccgg cattaaagtg accgttgaac atccggataa actggaagaa aaatttccgc
1621 aggttgcagc aaccggtgat ggtccggata ttatcttttg ggcacatgat cgttttggtg
1681 gttatgcaca gagcggtctg ctggcagaaa ttacaccgga taaagcattt caggacaaac
1741 tgtatccgtt tacctgggat gcagttcgct ataacggtaa actgattgca tatccgattg
1801 cagttgaagc actgagcctg atctataaca aagatctgct gccgaatccg cctaaaacct
1861 gggaagaaat tccggcactg gataaagaac tgaaagcaaa aggtaaaagc gcactgatgt
1921 ttaatctgca agaaccgtat tttacctggc ctctgattgc agcagatggt ggctatgcat
1981 tcaaatatga aaacggcaaa tatgatatca aagacgtggg tgttgataat gcgggtgcaa
2041 aagccggtct gacctttctg gttgatctga ttaaaaacaa acacatgaac gccgataccg
2101 attatagcat tgcagaagca gcatttaaca aaggtgaaac cgcaatgaca attaatggtc
2161 cgtgggcatg gtcaaatatt gataccagca aagtgaatta tggtgttacc gttctgccga
2221 catttaaagg tcagccgagc aaaccgtttg ttggtgttct gagcgcaggt attaatgcag
2281 caagcccgaa caaagaactg gcaaaagaat ttctggaaaa ctatctgctg accgatgaag
2341 gtctggaagc agtgaataaa gataaaccgc tgggtgcagt tgcactgaaa agctatgaag
2401 aagaactggt taaagatccg cgtattgcag ccaccatgga aaatgcacag aaaggcgaaa
2461 ttatgccgaa tattccgcag atgagcgcat tttggtatgc cgttcgtacc gcagtgatta
2521 atgccgcatc aggtcgtcag accgttgatg aagccctgaa agatgcccag accggtgcaa
2581 gcaccggatc cctggttccg cgtggtagcg ctagcgaaaa tctgtatttt cagggaatga
2641 gcaagctgga gaagtttaca aactgctact ccctgtctaa gaccctgagg ttcaaggcca
2701 tccctgtggg caagacccag gagaacatcg acaataagcg gctgctggtg gaggacgaga
2761 agagagccga ggattataag ggcgtgaaga agctgctgga tcgctactat ctgtctttta
2821 tcaacgacgt gctgcacagc atcaagctga agaatctgaa caattacatc agcctgttcc
2881 ggaagaaaac cagaaccgag aaggagaata aggagctgga gaacctggag atcaatctgc
2941 ggaaggagat cgccaaggcc ttcaagggca acgagggcta caagtccctg tttaagaagg
3001 atatcatcga gacaatcctg ccagagttcc tggacgataa ggacgagatc gccctggtga
3061 acagcttcaa tggctttacc acagccttca ccggcttctt tgataacaga gagaatatgt
3121 tttccgagga ggccaagagc acatccatcg ccttcaggtg tatcaacgag aatctgaccc
3181 gctacatctc taatatggac atcttcgaga aggtggacgc catctttgat aagcacgagg
3241 tgcaggagat caaggagaag atcctgaaca gcgactatga tgtggaggat ttctttgagg
3301 gcgagttctt taactttgtg ctgacacagg agggcatcga cgtgtataac gccatcatcg
3361 gcggcttcgt gaccgagagc ggcgagaaga tcaagggcct gaacgagtac atcaacctgt
3421 ataatcagaa aaccaagcag aagctgccta agtttaagcc actgtataag caggtgctga
3481 gcgatcggga gtctctgagc ttctacggcg agggctatac atccgatgag gaggtgctgg
3541 aggtgtttag aaacaccctg aacaagaaca gcgagatctt cagctccatc aagaagctgg
3601 agaagctgtt caagaatttt gacgagtact ctagcgccgg catctttgtg aagaacggcc
3661 ccgccatcag cacaatctcc aaggatatct tcggcgagtg gaacgtgatc cgggacaagt
3721 ggaatgccga gtatgacgat atccacctga agaagaaggc cgtggtgacc gagaagtacg
3781 aggacgatcg gagaaagtcc ttcaagaaga tcggctcctt ttctctggag cagctgcagg
3841 agtacgccga cgccgatctg tctgtggtgg agaagctgaa ggagatcatc atccagaagg
3901 tggatgagat ctacaaggtg tatggctcct ctgagaagct gttcgacgcc gattttgtgc
3961 tggagaagag cctgaagaag aacgacgccg tggtggccat catgaaggac ctgctggatt
4021 ctgtgaagag cttcgagaat tacatcaagg ccttctttgg cgagggcaag gagacaaaca
4081 gggacgagtc cttctatggc gattttgtgc tggcctacga catcctgctg aaggtggacc
4141 acatctacga tgccatccgc aattatgtga cccagaagcc ctactctaag gataagttca
4201 agctgtattt tcagaaccct cagttcatgg gcggctggga caaggataag gagacagact
4261 atcgggccac catcctgaga tacggctcca agtactatct ggccatcatg gataagaagt
4321 acgccaagtg cctgcagaag atcgacaagg acgatgtgaa cggcaattac gagaagatca
4381 actataagct gctgcccggc cctaataaga tgctgccaaa ggtgttcttt tctaagaagt
4441 ggatggccta ctataacccc agcgaggaca tccagaagat ctacaagaat ggcacattca
4501 agaagggcga tatgtttaac ctgaatgact gtcacaagct gatcgacttc tttaaggata
4561 gcatctcccg gtatccaaag tggtccaatg cctacgattt caacttttct gagacagaga
4621 agtataagga catcgccggc ttttacagag aggtggagga gcagggctat aaggtgagct
4681 tcgagtctgc cagcaagaag gaggtggata agctggtgga ggagggcaag ctgtatatgt
4741 tccagatcta taacaaggac ttttccgata agtctcacgg cacacccaat ctgcacacca
4801 tgtacttcaa gctgctgttt gacgagaaca atcacggaca gatcaggctg agcggaggag
4861 cagagctgtt catgaggcgc gcctccctga agaaggagga gctggtggtg cacccagcca
4921 actcccctat cgccaacaag aatccagata atcccaagaa aaccacaacc ctgtcctacg
4981 acgtgtataa ggataagagg ttttctgagg accagtacga gctgcacatc ccaatcgcca
5041 tcaataagtg ccccaagaac atcttcaaga tcaatacaga ggtgcgcgtg ctgctgaagc
5101 acgacgataa cccctatgtg atcggcatcg ataggggcga gcgcaatctg ctgtatatcg
5161 tggtggtgga cggcaagggc aacatcgtgg agcagtattc cctgaacgag atcatcaaca
5221 acttcaacgg catcaggatc aagacagatt accactctct gctggacaag aaggagaagg
5281 agaggttcga ggcccgccag aactggacct ccatcgagaa tatcaaggag ctgaaggccg
5341 gctatatctc tcaggtggtg cacaagatct gcgagctggt ggagaagtac gatgccgtga
5401 tcgccctgga ggacctgaac tctggcttta agaatagccg cgtgaaggtg gagaagcagg
5461 tgtatcagaa gttcgagaag atgctgatcg ataagctgaa ctacatggtg gacaagaagt
5521 ctaatccttg tgcaacaggc ggcgccctga agggctatca gatcaccaat aagttcgaga
5581 gctttaagtc catgtctacc cagaacggct tcatctttta catccctgcc tggctgacat
5641 ccaagatcga tccatctacc ggctttgtga acctgctgaa aaccaagtat accagcatcg
5701 ccgattccaa gaagttcatc agctcctttg acaggatcat gtacgtgccc gaggaggatc
5761 tgttcgagtt tgccctggac tataagaact tctctcgcac agacgccgat tacatcaaga
5821 agtggaagct gtactcctac ggcaaccgga tcagaatctt ccggaatcct aagaagaaca
5881 acgtgttcga ctgggaggag gtgtgcctga ccagcgccta taaggagctg ttcaacaagt
5941 acggcatcaa ttatcagcag ggcgatatca gagccctgct gtgcgagcag tccgacaagg
6001 ccttctactc tagctttatg gccctgatga gcctgatgct gcagatgcgg aacagcatca
6061 caggccgcac cgacgtggat tttctgatca gccctgtgaa gaactccgac ggcatcttct
6121 acgatagccg gaactatgag gcccaggaga atgccatcct gccaaagaac gccgacgcca
6181 atggcgccta taacatcgcc agaaaggtgc tgtgggccat cggccagttc aagaaggccg
6241 aggacgagaa gctggataag gtgaagatcg ccatctctaa caaggagtgg ctggagtacg
6301 cccagaccag cgtgaagcac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
6361 agaaaaaggg atcctaccca tacgatgttc cagattacgc ttatccctac gacgtgcctg
6421 attatgcata cccatatgat gtccccgact atgcctaaag cctcgagggt ggtggtagcg
6481 attataaaga tgatgatgat aaaggcctga acgatatctt tgaagcccag aaaattgaat
6541 ggcacgagta aaagcttgtc gagcaccacc accaccacca ctgagatccg gctgctaaca
6601 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
6661 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
6721 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
6781 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
6841 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
6901 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
6961 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
7021 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
7081 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
7141 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
7201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
7261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
7321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
7381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
7441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
7501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
7561 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
7621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
7681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
7741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
7801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
7861 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
7921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
7981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggtagtcgc
8041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
8101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
8161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
8221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
8281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
8341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
8401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
8461 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
8521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
8581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
8641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
8701 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
8761 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
8821 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
8881 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
8941 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
9001 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
9061 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
9121 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
9181 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
9241 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
9301 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
9361 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
9421 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
9481 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
9541 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
9601 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
9661 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
9721 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
9781 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
9841 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
9901 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
9961 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
10021 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
10081 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
10141 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
10201 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
10261 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
10321 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
10381 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
10441 tttgcgtatt gggcgccagg gtggttttt
//
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