| 质粒名称: | pC011 LwCas13a locus with targeting spacer |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 91903 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pACYC184 |
| 载体出品公司: | -- |
| 载体抗性: | -- |
| 空载体大小: | 4153 bp |
| 完整质粒大小: | 7791 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: | LwCas13a with targeting spacer |
| 物种来源: | -- |
| 插入大小: | 3456 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pC011 LwCas13a locus with targeting spacer was a gift from Feng Zhang (Addgene plasmid # 91903 ; http://n2t.net/addgene:91903 ; RRID:Addgene_91903) |
| 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 |
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 pC011 LwCas13a locus with targeting spacer 7790 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION LwCas13a locus into pACYC184 with targeting spacer.
ACCESSION .
VERSION .
KEYWORDS pC011 LwCas13a locus with targeting spacer
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7790)
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 7790)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7790
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 769..788
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter complement(1298..1332)
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
CDS complement(1336..4791)
/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"
promoter complement(4806..4835)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin 4873..5418
/direction=RIGHT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind 5319..5338
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5535..5552
/label=L4440
/note="L4440 vector, forward primer"
promoter 5944..6046
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 6047..6706
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(6113..6132)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS complement(7740..1140)
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
ORIGIN
1 gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc
61 ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc
121 gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
181 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
241 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
301 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
361 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
421 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
481 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
541 gactgggttg aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat
601 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
661 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
721 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
781 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
841 gtagaggatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag
901 tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg
961 tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat
1021 gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat
1081 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
1141 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttgtt
1201 ttagtcccct tcgtttttgg ggtagtctaa atctatggat tacttggtag aacagcaatc
1261 tgttttagtc cccttcgttt ttggggtagt ctaaatcgct agcattatac ctaggactga
1321 gctagctgtc aatcattcca gggccttgta ctcgaacatc actttcacga gttcgcacag
1381 ttcctcgctg ttccggtcgg tcatcagttt ctttttcttc agattcttca ggtgcacgat
1441 cttctctgat tccagggtct ggatttcgat cttcttgtca gcgccgatct tgaaggtggc
1501 cacgaagccg tattctttca gaatgtccac gatggacttc atgatggcgt tcttcagctt
1561 ccggtcgtag gacagcagct tccgcaggtt ttccagcact tccagcaggc taatctcggc
1621 gtgggggatg tagttgaagt gggcaatgta gttccggatg tacaggtcct ttttttcctg
1681 cttcagtttc ttcactttct tgtcggagta gatgctccgc ttttccacat tgtccttgta
1741 cagttctttg tagaagttga tatacttttc cacgatctgg ccgcttttgt acttcacatt
1801 cttggagttg tcgaaattga aaatttcctc gatgtagtgg ttctcgggaa actcgccctt
1861 cagccggaat ctcaggtccc gctcccagat gctggtgtag cccacgagcc ggtgcaggat
1921 cttcagcagc aggccctgca gcaggttcag ctcattgaat tccaccttgt tcttcaggtg
1981 ggtgtacttc tggatgttgc cgatggcctt ctcatactct ttgtagtcct cgtcgttgaa
2041 cttttcgtcc ttcttgggtc tggcgtactt ccggtgcagg ttctgctgca tggtgtagtt
2101 cttttcaatc tcattcttct tgttgctgta ctctttcagt tctttcaggc tgatcttata
2161 cttggcctta tcggcgatct tttccagcag attcagcatg ccgtatttct tgatattgta
2221 gaaggcccgg tgcttgatga tgttctcgcc gtcgaaatag atcttgttgg tgtcgaactt
2281 tttcagctct ttccggtcct tgattttgtt ttcgttgaag tccaggaact tgccgatctc
2341 gttggcttcc agctcgaagt cctcggtcac tctgttgttg tccaggttca gcaggttgat
2401 cagttccagc tcgtcgctga aggtttcttc tttgttggcg gactggtact tttccaggct
2461 gcccttcagg ttggtcagct ctttgtggtt cagcagcttc aggatcaggt aaaacatgtt
2521 cagattctcg gtgtacttca gaatcttccc cagcttgatc tcgcgcacga actcattgat
2581 ctcgtgaggg atttctttgt tccgattgtg cttctcatag ttcttcagga tcttgtcgta
2641 cttctctttg ttatcctttt tgatcttgat cttggagaag atgtcgttgt tgtcattgtt
2701 gttgttgctc tcgatatact tcagattgtt cttgttcagg tagtcgatga agcccttcag
2761 gaaaatctgc tgaataaagt cgatgtaggt attcttttcc tctttgtcct ggttgttgat
2821 catctctctg ctctggatga tggccaggta ttccacgggc acggttttct cgatgttctc
2881 gaacttctga tacttgtagt ggccggtttt ctggttccgc tgcttgttaa tcttgatcac
2941 ttcattggtg atcttaaaga acaccttgga gtttttcacg aacttgttca ggaactcgcc
3001 gtagtagata ttcttcagca ggtagatctg ggcgtccttc tcttctttgt ccttgggcac
3061 gctccaaaaa aacttcaggg tattccgcag gtcctcaatc ttgttgtaca gcttggtgaa
3121 gctgggcacg aaggggatgt ttttgttcac gaagttgaac ttggtattct tcaggtactt
3181 gatgatcaca tccttctcgt agtagttgaa cacgttggcg ctgttcagct gcttgaagat
3241 tttcagcttc agcttctttt cgttgatttc gttctgaaac atcttcttgg agatctcgct
3301 gggggcgata ttcttgaagg cgaagatgtc cttgccttcc agttccaggt tgaagtgcac
3361 gatgccgtgt ctgatgctgc tgatggcctc gtcgatgttg gcgaagaagt cctcgatctc
3421 gttcttgttg tccatgttga agtcgtagct gtagaacatc ttcagatttt ctttcacttc
3481 gttctgcttg ttctcattgt agatcttgtc cacctcgccg gacacgtatt tctcttcgcc
3541 cttgttgttc ttcacggtct tgccccgcat ccggccggtg atatcgttct cgttctcggt
3601 ttccaggatg ttcctcaggc tgaagtaggc cacgctggac acgccgatga tgtttctcag
3661 gaaggcctcg ttctgccggt tccgggcgat aaagtcggag gtggcgatct cgcccacttg
3721 cagatagtag ttgtacttgc cgcagttccg cacgtaggtg tccagcttgt tcagcagttt
3781 gttttcgatc agctttttca gattctggta ctcgaagatc cgcttgatct tatcgttgct
3841 gatgttgctc agccgcttgt acacgtagtt tttcagcagc tgggacatct cgatttccac
3901 gaagtggcag aaggcgtact taatattctt gtcgttcagt tcctctttgt ccaggtagta
3961 cttgtagaac acctggcttt tcttcagctc agacatgtcg gggatcttct caatcagctc
4021 tttgatgttg ttcacgttct ggatctcttc gtagataatc ttggcgaagt tctctttgtc
4081 gttcttccgg ccgatgatct tgtgatagta ctcgcggatc ttgtacttct cgtgcttctt
4141 gctgttctcg atcaggaaaa acagtttctc gatatcctct ttcttataca gcttgtcgaa
4201 ggcttcctgc acgttgttga tgtagtcgtt gcgcttggcg ctctctctgt agtagtcgta
4261 gatgatgttc cgcttgctct tgccgcccac tttttccacg ttgttctcgt tgatcttctg
4321 gtagttggcc ttgttctctt cgaagctgta cttcaggctg ttgatcttgt tcagcttggc
4381 ttccacgtcc ttccgaaaga tttccagttc ctcagagttc acgtcctcgt tcagcaggat
4441 cttcttcagc acggagaagc tgttcttgtt tttcaggtcg tactcgctga tgtcctcttc
4501 gctatagttc ttgtcctgca cggcgttctt ttctttccgg ttcttcagat acagcacgct
4561 gtccttcagg tgcagcacct tgttgctaaa gaacttcttc aggttctctc ttctgatccg
4621 gttctcttcc tcggaggcgt tgtcggggtt cttgatgtag atgtccagcc ggatgctcag
4681 cagctcgctc agtctctcgc tggtccggtt ttcctcgctg gtggacttca cgagcttgcc
4741 ctcttcgatg tacttcttgt ggctgatgcc gtcgaccttg gtcactttca tggataagac
4801 ctcctaacat acgagccgga agcataaagt gtaaatctag aaatatttta tctgattaat
4861 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
4921 aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg
4981 taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc
5041 gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt
5101 ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
5161 actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga
5221 gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga
5281 aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat
5341 agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg
5401 gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct
5461 tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc
5521 gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat
5581 tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc
5641 ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg
5701 cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct gatagaaaca
5761 gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt tggcagcatc
5821 acccgacgca ctttgcgccg aataaatacc tgtgacggaa gatcacttcg cagaataaat
5881 aaatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaca
5941 cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
6001 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
6061 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
6121 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
6181 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
6241 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
6301 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
6361 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
6421 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
6481 gtcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
6541 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
6601 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
6661 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
6721 ttattggtgc ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa
6781 tggcagaaat tcgaaagcaa attcgacccg gtcgtcggtt cagggcaggg tcgttaaata
6841 gccgcttatg tctattgctg gtttaccggt ttattgacta ccggaagcag tgtgaccgtg
6901 tgcttctcaa atgcctgagg ccagtttgct caggctctcc ccgtggaggt aataattgac
6961 gatatgatca tttattctgc ctcccagagc ctgataaaaa cggttagcgc ttcgttaata
7021 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
7081 tggtgcaggg cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
7141 tgggcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
7201 cgacgatagt catgccccgc gcccaccgga aggagctacc ggcagcggtg cggactgttg
7261 taactcagaa taagaaatga ggccgctcat ggcgttgact ctcagtcata gtatcgtggt
7321 atcaccggtt ggttccactc tctgttgcgg gcaacttcag cagcacgtag gggacttccg
7381 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
7441 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
7501 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
7561 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
7621 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
7681 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
7741 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
//
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