| 质粒名称: | pC019 - LwCas13a from Leptotrichia wadei |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 91909 |
| 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: | 4197 bp |
| 完整质粒大小: | 7783 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 and guide expression cassette |
| 物种来源: | Leptotrichia wadei |
| 插入大小: | 3586 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: | pC019 - LwCas13a from Leptotrichia wadei was a gift from Feng Zhang (Addgene plasmid # 91909 ; http://n2t.net/addgene:91909 ; RRID:Addgene_91909) |
| 参考文献部分: | RNA targeting with CRISPR-Cas13. Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F. Nature. 2017 Oct 4. doi: 10.1038/nature24049. 10.1038/nature24049 PubMed 28976959 |
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 pC019 - LwCas13a from Leptotrichia wadei 7782 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses LwCas13a for bacterial expression and contains backbone
for spacer cloning.
ACCESSION .
VERSION .
KEYWORDS pC019 - LwCas13a from Leptotrichia wadei
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7782)
AUTHORS Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J,
Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES,
Voytas DF, Ting AY, Zhang F
TITLE RNA targeting with CRISPR-Cas13.
JOURNAL Nature. 2017 Oct 4. doi: 10.1038/nature24049.
PUBMED 28976959
REFERENCE 2 (bases 1 to 7782)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7782
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 55..84
/label=lac promoter
/note="promoter for the E. coli lac operon"
CDS 99..3554
/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 3558..3592
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
CDS 3742..4932
/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 complement(4094..4113)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
CDS complement(5966..6625)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 6540..6559
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(6626..6728)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(7120..7137)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7254..17)
/direction=LEFT
/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 complement(7334..7353)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttaca
61 ctttatgctt ccggctcgta tgttaggagg tcttatccat gaaagtgacc aaggtcgacg
121 gcatcagcca caagaagtac atcgaagagg gcaagctcgt gaagtccacc agcgaggaaa
181 accggaccag cgagagactg agcgagctgc tgagcatccg gctggacatc tacatcaaga
241 accccgacaa cgcctccgag gaagagaacc ggatcagaag agagaacctg aagaagttct
301 ttagcaacaa ggtgctgcac ctgaaggaca gcgtgctgta tctgaagaac cggaaagaaa
361 agaacgccgt gcaggacaag aactatagcg aagaggacat cagcgagtac gacctgaaaa
421 acaagaacag cttctccgtg ctgaagaaga tcctgctgaa cgaggacgtg aactctgagg
481 aactggaaat ctttcggaag gacgtggaag ccaagctgaa caagatcaac agcctgaagt
541 acagcttcga agagaacaag gccaactacc agaagatcaa cgagaacaac gtggaaaaag
601 tgggcggcaa gagcaagcgg aacatcatct acgactacta cagagagagc gccaagcgca
661 acgactacat caacaacgtg caggaagcct tcgacaagct gtataagaaa gaggatatcg
721 agaaactgtt tttcctgatc gagaacagca agaagcacga gaagtacaag atccgcgagt
781 actatcacaa gatcatcggc cggaagaacg acaaagagaa cttcgccaag attatctacg
841 aagagatcca gaacgtgaac aacatcaaag agctgattga gaagatcccc gacatgtctg
901 agctgaagaa aagccaggtg ttctacaagt actacctgga caaagaggaa ctgaacgaca
961 agaatattaa gtacgccttc tgccacttcg tggaaatcga gatgtcccag ctgctgaaaa
1021 actacgtgta caagcggctg agcaacatca gcaacgataa gatcaagcgg atcttcgagt
1081 accagaatct gaaaaagctg atcgaaaaca aactgctgaa caagctggac acctacgtgc
1141 ggaactgcgg caagtacaac tactatctgc aagtgggcga gatcgccacc tccgacttta
1201 tcgcccggaa ccggcagaac gaggccttcc tgagaaacat catcggcgtg tccagcgtgg
1261 cctacttcag cctgaggaac atcctggaaa ccgagaacga gaacgatatc accggccgga
1321 tgcggggcaa gaccgtgaag aacaacaagg gcgaagagaa atacgtgtcc ggcgaggtgg
1381 acaagatcta caatgagaac aagcagaacg aagtgaaaga aaatctgaag atgttctaca
1441 gctacgactt caacatggac aacaagaacg agatcgagga cttcttcgcc aacatcgacg
1501 aggccatcag cagcatcaga cacggcatcg tgcacttcaa cctggaactg gaaggcaagg
1561 acatcttcgc cttcaagaat atcgccccca gcgagatctc caagaagatg tttcagaacg
1621 aaatcaacga aaagaagctg aagctgaaaa tcttcaagca gctgaacagc gccaacgtgt
1681 tcaactacta cgagaaggat gtgatcatca agtacctgaa gaataccaag ttcaacttcg
1741 tgaacaaaaa catccccttc gtgcccagct tcaccaagct gtacaacaag attgaggacc
1801 tgcggaatac cctgaagttt ttttggagcg tgcccaagga caaagaagag aaggacgccc
1861 agatctacct gctgaagaat atctactacg gcgagttcct gaacaagttc gtgaaaaact
1921 ccaaggtgtt ctttaagatc accaatgaag tgatcaagat taacaagcag cggaaccaga
1981 aaaccggcca ctacaagtat cagaagttcg agaacatcga gaaaaccgtg cccgtggaat
2041 acctggccat catccagagc agagagatga tcaacaacca ggacaaagag gaaaagaata
2101 cctacatcga ctttattcag cagattttcc tgaagggctt catcgactac ctgaacaaga
2161 acaatctgaa gtatatcgag agcaacaaca acaatgacaa caacgacatc ttctccaaga
2221 tcaagatcaa aaaggataac aaagagaagt acgacaagat cctgaagaac tatgagaagc
2281 acaatcggaa caaagaaatc cctcacgaga tcaatgagtt cgtgcgcgag atcaagctgg
2341 ggaagattct gaagtacacc gagaatctga acatgtttta cctgatcctg aagctgctga
2401 accacaaaga gctgaccaac ctgaagggca gcctggaaaa gtaccagtcc gccaacaaag
2461 aagaaacctt cagcgacgag ctggaactga tcaacctgct gaacctggac aacaacagag
2521 tgaccgagga cttcgagctg gaagccaacg agatcggcaa gttcctggac ttcaacgaaa
2581 acaaaatcaa ggaccggaaa gagctgaaaa agttcgacac caacaagatc tatttcgacg
2641 gcgagaacat catcaagcac cgggccttct acaatatcaa gaaatacggc atgctgaatc
2701 tgctggaaaa gatcgccgat aaggccaagt ataagatcag cctgaaagaa ctgaaagagt
2761 acagcaacaa gaagaatgag attgaaaaga actacaccat gcagcagaac ctgcaccgga
2821 agtacgccag acccaagaag gacgaaaagt tcaacgacga ggactacaaa gagtatgaga
2881 aggccatcgg caacatccag aagtacaccc acctgaagaa caaggtggaa ttcaatgagc
2941 tgaacctgct gcagggcctg ctgctgaaga tcctgcaccg gctcgtgggc tacaccagca
3001 tctgggagcg ggacctgaga ttccggctga agggcgagtt tcccgagaac cactacatcg
3061 aggaaatttt caatttcgac aactccaaga atgtgaagta caaaagcggc cagatcgtgg
3121 aaaagtatat caacttctac aaagaactgt acaaggacaa tgtggaaaag cggagcatct
3181 actccgacaa gaaagtgaag aaactgaagc aggaaaaaaa ggacctgtac atccggaact
3241 acattgccca cttcaactac atcccccacg ccgagattag cctgctggaa gtgctggaaa
3301 acctgcggaa gctgctgtcc tacgaccgga agctgaagaa cgccatcatg aagtccatcg
3361 tggacattct gaaagaatac ggcttcgtgg ccaccttcaa gatcggcgct gacaagaaga
3421 tcgaaatcca gaccctggaa tcagagaaga tcgtgcacct gaagaatctg aagaaaaaga
3481 aactgatgac cgaccggaac agcgaggaac tgtgcgaact cgtgaaagtg atgttcgagt
3541 acaaggccct ggaatgattg acagctagct cagtcctagg tataatgcta gcgatttaga
3601 ctaccccaaa aacgaagggg actaaaacgg agacggagat tcgtctcgga tttagactac
3661 cccaaaaacg aaggggacta aaacaagctt taatgcggta gtttatcaca gttaaattgc
3721 taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc tcatcgtcat cctcggcacc
3781 gtcaccctgg atgctgtagg cataggcttg gttatgccgg tactgccggg cctcttgcgg
3841 gatatcgtcc attccgacag catcgccagt cactatggcg tgctgctagc gctatatgcg
3901 ttgatgcaat ttctatgcgc acccgttctc ggagcactgt ccgaccgctt tggccgccgc
3961 ccagtcctgc tcgcttcgct acttggagcc actatcgact acgcgatcat ggcgaccaca
4021 cccgtcctgt ggatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
4081 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
4141 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
4201 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
4261 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg accgatgccc
4321 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
4381 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
4441 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
4501 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
4561 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
4621 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
4681 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
4741 ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg
4801 atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta
4861 ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc
4921 acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg
4981 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
5041 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct
5101 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
5161 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
5221 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
5281 gtctggaaac gcggaagtcc cctacgtgct gctgaagttg cccgcaacag agagtggaac
5341 caaccggtga taccacgata ctatgactga gagtcaacgc catgagcggc ctcatttctt
5401 attctgagtt acaacagtcc gcaccgctgc cggtagctcc ttccggtggg cgcggggcat
5461 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
5521 ggcagcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
5581 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
5641 acctacatct gtattaacga agcgctaacc gtttttatca ggctctggga ggcagaataa
5701 atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg gcctcaggca
5761 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
5821 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
5881 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag
5941 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
6001 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
6061 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
6121 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
6181 gattggctga cacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
6241 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
6301 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
6361 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag
6421 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
6481 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
6541 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
6601 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
6661 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
6721 gccgatcaac gtgtcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
6781 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa
6841 gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct
6901 ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt
6961 aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg
7021 gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt
7081 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
7141 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
7201 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
7261 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
7321 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
7381 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
7441 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
7501 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
7561 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
7621 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
7681 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
7741 gcaaggcggt tttttcgttt tcagagcaag agattacgcg ca
//
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