| 质粒名称: | pC013 - Twinstrep-SUMO-huLwCas13a |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 90097 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pET |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 6068 bp |
| 完整质粒大小: | 9542 bp |
| 载体类型: | 细菌表达 |
| 宿生菌： | Rosetta 2(DE3)pLysS Singles |
| 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: | LwCas13a |
| 备用名称： | LwC2c2、Cas13a、C2c2 |
| 物种来源: | 合成的；韦氏钩端螺旋体 |
| 插入大小: | 3280 bp |
| 标签/融合蛋白: | 6x His Tag/Twin Strep/SUMO (N terminal on insert) |
|
文献引用方法 |
|
| 材料和方法部分: | pC013 - Twinstrep-SUMO-huLwCas13a was a gift from Feng Zhang (Addgene plasmid # 90097 ; http://n2t.net/addgene:90097 ; RRID:Addgene_90097) |
| 参考文献部分: |
Nucleic acid detection with CRISPR-Cas13a/C2c2. Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC, Collins JJ, Zhang F. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321. 10.1126/science.aam9321 PubMed 28408723
Related items:
From this article
Feng Zhang Lab Plasmids
CRISPR/Cas Items
Commonly requested with:
pC015 - dLwCas13a-NF
pC016 - LwCas13a guide expression backbone with U6 promoter
pC001 - huLshC2C2-MBP for bacterial expression
pC017 - LwCas13a guide expression backbone with tRNAval promoter
p2CT-His-MBP-Lwa_Cas13a_WT
|
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 pC013 - Twinstrep-SUMO-huLwCas13a 9542 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Twinstrep-SUMO-huLwCas13a for recombinant protein bacterial
expression. Insert is human codon optimized but expresses well in
bacteria..
ACCESSION .
VERSION .
KEYWORDS pC013 - Twinstrep-SUMO-huLwCas13a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9542)
AUTHORS Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung
J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C,
Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC,
Collins JJ, Zhang F
TITLE Nucleic acid detection with CRISPR-Cas13a/C2c2.
JOURNAL Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
PUBMED 28408723
REFERENCE 2 (bases 1 to 9542)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9542
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(170..192)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 278..417
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 333..352
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(432..449)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(603..1191)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(683..702)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1362..2222)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 1985..2004
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2223..2327)
/gene="bla"
/label=AmpR promoter
primer_bind 2395..2413
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2440..2468
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
terminator 2643..2690
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 2686..2704
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(2754..6209)
/codon_start=1
/product="Leptotrichia wadei CRISPR-Cas effector that acts
as an RNA-guided RNAse (Abudayyeh et al., 2017)"
/label=LwCas13a
/note="previously called C2c2"
/translation="MKVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIY
IKNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEY
DLKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINEN
NVEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEK
YKIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLD
KEELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLL
NKLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETE
NENDITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNE
IEDFFANIDEAISSIRHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLK
IFKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFW
SVPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQ
KFENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIE
SNNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKY
TENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDF
ELEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEK
IADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAI
GNIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEE
IFNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIRNY
IAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKK
IEIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
CDS complement(6216..6506)
/codon_start=1
/gene="S. cerevisiae SMT3 (truncated)"
/product="cleavable ubiquitin-like protein tag"
/label=SUMO
/translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
CDS complement(6507..6530)
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS complement(6567..6590)
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS complement(6603..6620)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(6630..6647)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 6666..6688
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 6703..6727
/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(6727..6746)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6728..6746)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6830..6849)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 7055..7132
/gene="lacI"
/label=lacI promoter
CDS 7133..8215
/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(7152..7171)
/label=LacI-R
/note="LacI, reverse primer"
CDS 9527..176
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
61 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
121 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
301 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
361 gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
421 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
481 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
541 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
1021 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
1081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
1141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
1201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
1261 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
1321 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
1381 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
1441 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
1501 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
1561 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
1621 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
1681 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
1741 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
1801 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
1861 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
1921 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
1981 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
2041 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
2101 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
2161 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
2221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
2281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
2341 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
2401 cgtatcacga ggccctttcg tcttcaagaa ttctcatgtt tgacagctta tcatcgataa
2461 gctttaatgc ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa
2521 atctaacaat gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg
2581 cttggttatg ccggtactgc cgggcctctt gcgggatatc cggatatagt tcctcctttc
2641 agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag ttattgctca
2701 gcggtggcag cagccaactc agcttccttt cgggcctcga gtgcggccgc ttattccagg
2761 gccttgtact cgaacatgac tttcacgagt tcgcacagtt cctcgctgtt ccggtcggtc
2821 atcagtttct ttttcttcag attcttcagg tgcacgatct tctctgattc cagggtctgg
2881 atttcgatct tcttgtcagc gccgatcttg aaggtggcca cgaagccgta ttctttcaga
2941 atgtccacga tggacttcat gatggcgttc ttcagcttcc ggtcgtagga cagcagcttc
3001 cgcaggtttt ccagcacttc cagcaggcta atctcggcgt gggggatgta gttgaagtgg
3061 gcaatgtagt tccggatgta caggtccttt ttttcctgct tcagtttctt cactttcttg
3121 tcggagtaga tgctccgctt ttccacattg tccttgtaca gttctttgta gaagttgata
3181 tacttttcca cgatctggcc gcttttgtac ttcacattct tggagttgtc gaaattgaaa
3241 atttcctcga tgtagtggtt ctcgggaaac tcgcccttca gccggaatct caggtcccgc
3301 tcccagatgc tggtgtagcc cacgagccgg tgcaggatct tcagcagcag gccctgcagc
3361 aggttcagct cattgaattc caccttgttc ttcaggtggg tgtacttctg gatgttgccg
3421 atggccttct catactcttt gtagtcctcg tcgttgaact tttcgtcctt cttgggtctg
3481 gcgtacttcc ggtgcaggtt ctgctgcatg gtgtagttct tttcaatctc attcttcttg
3541 ttgctgtact ctttcagttc tttcaggctg atcttatact tggccttatc ggcgatcttt
3601 tccagcagat tcagcatgcc gtatttcttg atattgtaga aggcccggtg cttgatgatg
3661 ttctcgccgt cgaaatagat cttgttggtg tcgaactttt tcagctcttt ccggtccttg
3721 attttgtttt cgttgaagtc caggaacttg ccgatctcgt tggcttccag ctcgaagtcc
3781 tcggtcactc tgttgttgtc caggttcagc aggttgatca gttccagctc gtcgctgaag
3841 gtttcttctt tgttggcgga ctggtacttt tccaggctgc ccttcaggtt ggtcagctct
3901 ttgtggttca gcagcttcag gatcaggtaa aacatgttca gattctcggt gtacttcaga
3961 atcttcccca gcttgatctc gcgcacgaac tcattgatct cgtgagggat ttctttgttc
4021 cgattgtgct tctcatagtt cttcaggatc ttgtcgtact tctctttgtt atcctttttg
4081 atcttgatct tggagaagat gtcgttgttg tcattgttgt tgttgctctc gatatacttc
4141 agattgttct tgttcaggta gtcgatgaag cccttcagga aaatctgctg aataaagtcg
4201 atgtaggtat tcttttcctc tttgtcctgg ttgttgatca tctctctgct ctggatgatg
4261 gccaggtatt ccacgggcac ggttttctcg atgttctcga acttctgata cttgtagtgg
4321 ccggttttct ggttccgctg cttgttaatc ttgatcactt cattggtgat cttaaagaac
4381 accttggagt ttttcacgaa cttgttcagg aactcgccgt agtagatatt cttcagcagg
4441 tagatctggg cgtccttctc ttctttgtcc ttgggcacgc tccaaaaaaa cttcagggta
4501 ttccgcaggt cctcaatctt gttgtacagc ttggtgaagc tgggcacgaa ggggatgttt
4561 ttgttcacga agttgaactt ggtattcttc aggtacttga tgatcacatc cttctcgtag
4621 tagttgaaca cgttggcgct gttcagctgc ttgaagattt tcagcttcag cttcttttcg
4681 ttgatttcgt tctgaaacat cttcttggag atctcgctgg gggcgatatt cttgaaggcg
4741 aagatgtcct tgccttccag ttccaggttg aagtgcacga tgccgtgtct gatgctgctg
4801 atggcctcgt cgatgttggc gaagaagtcc tcgatctcgt tcttgttgtc catgttgaag
4861 tcgtagctgt agaacatctt cagattttct ttcacttcgt tctgcttgtt ctcattgtag
4921 atcttgtcca cctcgccgga cacgtatttc tcttcgccct tgttgttctt cacggtcttg
4981 ccccgcatcc ggccggtgat atcgttctcg ttctcggttt ccaggatgtt cctcaggctg
5041 aagtaggcca cgctggacac gccgatgatg tttctcagga aggcctcgtt ctgccggttc
5101 cgggcgataa agtcggaggt ggcgatctcg cccacttgca gatagtagtt gtacttgccg
5161 cagttccgca cgtaggtgtc cagcttgttc agcagtttgt tttcgatcag ctttttcaga
5221 ttctggtact cgaagatccg cttgatctta tcgttgctga tgttgctcag ccgcttgtac
5281 acgtagtttt tcagcagctg ggacatctcg atttccacga agtggcagaa ggcgtactta
5341 atattcttgt cgttcagttc ctctttgtcc aggtagtact tgtagaacac ctggcttttc
5401 ttcagctcag acatgtcggg gatcttctca atcagctctt tgatgttgtt cacgttctgg
5461 atctcttcgt agataatctt ggcgaagttc tctttgtcgt tcttccggcc gatgatcttg
5521 tgatagtact cgcggatctt gtacttctcg tgcttcttgc tgttctcgat caggaaaaac
5581 agtttctcga tatcctcttt cttatacagc ttgtcgaagg cttcctgcac gttgttgatg
5641 tagtcgttgc gcttggcgct ctctctgtag tagtcgtaga tgatgttccg cttgctcttg
5701 ccgcccactt tttccacgtt gttctcgttg atcttctggt agttggcctt gttctcttcg
5761 aagctgtact tcaggctgtt gatcttgttc agcttggctt ccacgtcctt ccgaaagatt
5821 tccagttcct cagagttcac gtcctcgttc agcaggatct tcttcagcac ggagaagctg
5881 ttcttgtttt tcaggtcgta ctcgctgatg tcctcttcgc tatagttctt gtcctgcacg
5941 gcgttctttt ctttccggtt cttcagatac agcacgctgt ccttcaggtg cagcaccttg
6001 ttgctaaaga acttcttcag gttctctctt ctgatccggt tctcttcctc ggaggcgttg
6061 tcggggttct tgatgtagat gtccagccgg atgctcagca gctcgctcag tctctcgctg
6121 gtccggtttt cctcgctggt ggacttcacg agcttgccct cttcgatgta cttcttgtgg
6181 ctgatgccgt cgaccttggt cactttcatg gatccaccaa tctgttctct gtgagcctca
6241 ataatatcgt tatcctccat gtccaaatct tcaggggtct gatcagcttg gattctaata
6301 ccgtcgtaca agaatcttaa ggagtccatt tccttaccct gtcttttagc gaacgcttcc
6361 atcagccttc ttaaaggagt ggtctttttg atcttgaaga agatctctga agatccatcg
6421 gacaccttta aattgatgtg agtctcaggc ttgacttctg gcttgacctc tggcttagct
6481 tcttgattga cttctgagtc cgacattttc tcaaactgag ggtgtgacca tgcactacca
6541 cctgaaccac caccgctacc accacctttt tcaaactgcg gatggctcca gctagccata
6601 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
6661 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
6721 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
6781 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
6841 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
6901 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
6961 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
7021 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
7081 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
7141 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
7201 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
7261 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
7321 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
7381 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
7441 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
7501 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
7561 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
7621 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
7681 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
7741 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
7801 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
7861 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
7921 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
7981 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
8041 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
8101 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
8161 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
8221 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
8281 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
8341 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
8401 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
8461 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
8521 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
8581 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
8641 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
8701 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga ccgctgatcg
8761 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg
8821 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
8881 cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc
8941 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
9001 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc
9061 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
9121 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
9181 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
9241 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
9301 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
9361 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
9421 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
9481 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
9541 aa
//
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