| 质粒名称: | pC009 LshCas13a locus with targeting spacer |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 91900 |
| 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: | 氯霉素 |
| 空载体大小: | 4153 bp |
| 完整质粒大小: | 8486 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: | LshCas13a locus with targeting spacer |
| 物种来源: | leptotrichia shahii |
| 插入大小: | 4333 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 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: | pC009 LshCas13a locus with targeting spacer was a gift from Feng Zhang (Addgene plasmid # 91900 ; http://n2t.net/addgene:91900 ; RRID:Addgene_91900) |
| 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 |
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载体基本信息
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载体简介
载体序列
LOCUS pC009 LshCas13a locus with targeting spacer 8485 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION LshCas13a locus into pACYC184 with targeting spacer.
ACCESSION .
VERSION .
KEYWORDS pC009 LshCas13a locus with targeting spacer
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8485)
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 8485)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8485
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(390..424)
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
promoter complement(4609..4638)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin 4676..5221
/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 5122..5141
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5338..5355
/label=L4440
/note="L4440 vector, forward primer"
promoter 5747..5849
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 5850..6509
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(5916..5935)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS complement(7543..248)
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
primer_bind 8362..8381
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
61 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
121 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
181 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
241 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
301 agcttgtttt agtccccttc gatattgggg tggtatggat tacttggtag aacagcaatc
361 tgttttagtc cccttcgata ttggggtggg ctagcactat acctaggact gagctagctg
421 tcaatcacag ggtatcgtta gtattctcga ttttggtcag cagttcaatg atcagattct
481 taatgtagtc actgttgtag gactccagtt ccaggacgct cactttcttg ggcttcatca
541 ggcgctccag aatatcattg ttgccgatca gtttgaattt ctttttcagc tcatcgtagt
601 ccagattcac gtccttttta aagacttcga acacgctggc ataagttgaa ttgttgtagc
661 gtgtgctata tgacagcagg ttgctcaccc gatcgatctg ctcagcaata ctgtagtcgg
721 caaatggatt tctcacgatg tagaaatggg agatatagtt ccgaatgctc tcattttcag
781 gcttgttgat ctcggaattt tcgctcaggt cgattccaaa cccgtagcaa atcttctcga
841 acttcttgta ggactcttca tcaaagaact tgtagtaggc tgtggtagtg taaaagccgt
901 cgcttccatt gcgctttggg tatgccctgc tgatcccggt gttatagcct gacagcttaa
961 tgatgcccag ctccctcagt ccattcacga tatagtgcat atccctttcg aatcttgcca
1021 tctgaatagc cagcttccag ttgatgtcaa tcaggtagga ctcgattttg ttcagatagt
1081 tgaactcgac cagatctcga atctttttgt actcagacac gcggttatag tccttttcga
1141 aggatttgta gttcttattc tggatgttct ttgcaaagaa atcgtcgttc tccttcagct
1201 ttttgatgta cttctccttg tattcctttg agtagccgtt cagtttatcg ttcagattct
1261 tcagaatagc gtcgatctca ctgattttgt tctttctgat gttctttcca tcgatattaa
1321 acaggaattt agcatcggcc atcttgatgt cgttgctaat caggccgtag atcttgtcaa
1381 agttaggatt tccgatgttc aggaacagat tctttttgta gatatacagc tcgttcttcc
1441 gttccttggg atagtagatc tcctggaact tgttctcatt ttcagactcc atatcctcaa
1501 tcagattgtc gatttccttc ttgtactttt tcagaaagtc actgttgaaa atgatcctgc
1561 acaggatttt agacttgatc tcctgatctt tgtccttgat atactgatcc acctttttct
1621 tcaggtcctt cttgttgatg ttagacagct ttctctgttc atcctgcagg atgttggatt
1681 tcttgtcgat ctcgaacttg gtttcatcgt cgaagatgac aatcttctcc agtttctttt
1741 ccagcacatc acagccgttg atatcgtctt taaactcggt cagaatgttg ttcttgatgt
1801 cctcgtagta gttgttgaag atttctttct tagtctggat cttgaagtcg tcgaaatcct
1861 tttcgatctc tttcatcttc tgaatgaact cttccaggtt cagattccag ttctctgtga
1921 tgcattcgtt ccgcagggta ttcagctgca taatctcatc caggatgtca atgatattct
1981 ggtattctga ggtgttcagc cacacgctgg tggcgaagaa tctgtttcgg atcttattaa
2041 tcacggcatt actgttcagc agtgcaaaga tagaaatgat gtactcgaaa tcgtcgttga
2101 tgacaatggt tttgtcgctg gttttcacgg tgatccactc ataagtcttg ttatcgttga
2161 tgtccttaat ctgtttcttg atttcctgga tgttcatctt gaagtctgaa aagtcgaaca
2221 gctcttcgta gtttttgcgc agatatccaa tgtagcactc gatcactttc ttctgatatt
2281 tcttgattgc cttgttgttc cccttactgg cagaaatctg tgcgttcttg tagtagttct
2341 cgatgatgtt ctcatcgatt tcgtcaatgt tgcccagggt tttcttcagt tcctgcagga
2401 agatattctt ggactcgttc tcttccagat cgtcctccag gatcagtttc ttatacagct
2461 ccttattgac gtagatcaga gcgttcagca caattttttc agtctcgatt gtgtcaaaag
2521 gctcattctt gggattgttg cggtacaggt tcaggatttc gggcagcact ttgctgaagc
2581 tgggcagata cttgatatcg ttgttgttct cttcagagat cttgatgtca ttgatctttg
2641 tgatgatgtt tttcttgtcc ttaaagacca cgtccaggtt cagagctttg ctcacctctt
2701 catcggagat cttcagattc tgaatgatgt tgatcacttt gttgtaatcg tcctgagtcc
2761 cctgcaggtc cctctctttg gaaatggcat gcaggatgcg gttcctttca ttagtgccga
2821 tctttgtaaa ctttcgaatg aagttgtttg tgatgttgtt cttgttatcg atgaaatcca
2881 ggtcgcggat gatcttgatc ttactattca ggattttctt atccagcaca tagttcttct
2941 cccggtcgcc tccaaagaaa tcgatgttct cgtcgttgtt gatgttttct cgggaaaaga
3001 tcttgttcag ctccatattt gtgctggcaa agaaggtgat cagttccagg tccagctctt
3061 ccttggcgtg caggcggctg aaatcgtcag tattcactgt ggtcatatcg atgtcgttat
3121 gcctcagctt ccccaggtac ataatgtgct ccagtgtata ctgtttcact ctcttcagaa
3181 ttttctctga caggatactt tcattcagaa tcttttcgat ctcgatcttc tccattttct
3241 tcagtctgac cttctgctcg ttcaccagga ttttttcaat tctgcccttc aggtatctgt
3301 agatgatctt atacagctcc ttctcttcat cagatttctt ggaaaatttc ttgctgtcga
3361 agttcacctt gtaatgcttc ttaaagattc cgaagatctc ggtatcacag ttccctttct
3421 tcagttcctt ctccagtttc ttaatcagct cgtcgatctt gaactcagcc aggatcttct
3481 caatcttttc cttgatagag ttgttcttaa tgttctcgac aaagaacttc acgatcttat
3541 ccttcttgtt ttctctctcg attttaaact tttcgtgctt gtccagcagc acataggact
3601 tgatgtatgt ccgatttctc cgcttctcca ggaactcgtt attgactttc ttcactttct
3661 caatgcgctt tgtgatgttc caaaattcca gctccttgat cacgaaatcg gcaatgtcct
3721 cgacggtcag atccacattg atgttcagaa tcttttccac cagcattttc ttatttttac
3781 ttttcttttt gtccccgccg acgttcaggt aaaacttcac gaagcccagg atctccagat
3841 tagacttgat cttctcccgg atttccatga agttggtcag gatcacgtca atcttatcgt
3901 ctttcagcag cttctccctc aggtgctctt cgtagtatct attttcaaac accttttctg
3961 tctcgttttc gatgatcttc tcgatgatct tatacaggct catattgatg ttcttgaaga
4021 tctcgtagat gctcttcttt gtctccagtt catcgttttc aatgatgcgc agaatgatgg
4081 agcagtcgtt cagggtctta ttagtgtact catcccggat gtcaatttcg atctcttcct
4141 cgttctcctg tcgcttgatc tcgatctttt tatcgtcctt agtaatgccc tgcctgatgg
4201 cctcgtcaat gatctttttc tttgtgatcc ccagtgcttt cagcttctcg ctcttgccgt
4261 aggcctcgat atacagcacc acttcctctg tctccaggaa gtcgtcgtta ttctcaattc
4321 taatgatccc ttctttgccc ttcagtttga acaggatatt tcctgcatga aatttcctag
4381 tgaactcctt cagaatatta tcgtttttct tgtagttgat gtactttctg atgaacttgt
4441 tgttgtcgat cttctctttg ttgttgtttt cgttgatgtt cagaatgtac ttgttcccgt
4501 cataattccg cttgactttc acctttcgct tgatcttgaa gtctttctta tctcgcacct
4561 cgtaccaccg cttgtgtccg aacaggttcc ccatggataa gacctcctaa catacgagcc
4621 ggaagcataa agtgtaaatc tagaaatatt ttatctgatt aataagatga tcttcttgag
4681 atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg
4741 gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag
4801 cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta
4861 actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg
4921 ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg
4981 tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg
5041 agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga
5101 gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
5161 ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa
5221 aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg
5281 aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc
5341 gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt
5401 gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat
5461 agtaagccag tatacactcc gctagcgctg atgtccggcg gtgcttttgc cgttacgcac
5521 caccccgtca gtagctgaac aggagggaca gctgatagaa acagaagcca ctggagcacc
5581 tcaaaaacac catcatacac taaatcagta agttggcagc atcacccgac gcactttgcg
5641 ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg
5701 ttgataccgg gaagccctgg gccaactttt ggcgaaaatg acacgttgat cggcacgtaa
5761 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
5821 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
5881 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
5941 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
6001 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
6061 cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
6121 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
6181 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
6241 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtgtcagcc aatccctggg
6301 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
6361 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
6421 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
6481 actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa
6541 cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag
6601 caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg
6661 ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
6721 aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga tcatttattc
6781 tgcctcccag agcctgataa aaacggttag cgcttcgtta atacagatgt aggtgttcca
6841 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgcttgt
6901 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc tgccggcacc
6961 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
7021 cgcgcccacc ggaaggagct accggcagcg gtgcggactg ttgtaactca gaataagaaa
7081 tgaggccgct catggcgttg actctcagtc atagtatcgt ggtatcaccg gttggttcca
7141 ctctctgttg cgggcaactt cagcagcacg taggggactt ccgcgtttcc agactttacg
7201 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
7261 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
7321 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc
7381 caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt
7441 tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc
7501 ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg
7561 gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat
7621 ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg
7681 tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg
7741 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
7801 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
7861 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
7921 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
7981 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
8041 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
8101 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
8161 cggtcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
8221 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
8281 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
8341 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
8401 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atccacagga cgggtgtggt
8461 cgccatgatc gcgtagtcga tagtg
//
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