| 质粒名称: | pYPQ230 (LbCpf1) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 86210 |
| 存储实验室: | Yiping Qi |
|
载体骨架基本信息 |
|
| 载体名称: | pYPQ167 |
| 载体抗性: | 大观霉素 |
| 载体类型: | 植物表达，CRISPR；网关兼容LbCpf1入口克隆 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | LbCpf1 |
| 物种来源: | Lachnospiraceae bacterium ND2006 |
| 标签/融合蛋白: | 5' and 3' NLS |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NcoI HindIII（销毁后未知） |
| 3' 克隆位点: | NheI-AatII（销毁后未知） |
| 5' 测序引物 | gtaaaacgacggccag |
|
文献引用方法 |
|
| 材料和方法部分: | pYPQ230 (LbCpf1) was a gift from Yiping Qi (Addgene plasmid # 86210 ; http://n2t.net/addgene:86210 ; RRID:Addgene_86210) |
| 参考文献部分: | A CRISPR-Cpf1 system for efficient genome editing and transcriptional repression in plants. Tang X, Lowder LG, Zhang T, Malzahn AA, Zheng X, Voytas DF, Zhong Z, Chen Y, Ren Q, Li Q, Kirkland ER, Zhang Y, Qi Y. Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18. 10.1038/nplants.2017.18 PubMed 28211909 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pYPQ230 (LbCpf1) 6939 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION LbCpf1 Gateway entry plasmid.
ACCESSION .
VERSION .
KEYWORDS pYPQ230 (LbCpf1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6939)
AUTHORS Tang X, Lowder LG, Zhang T, Malzahn AA, Zheng X, Voytas DF, Zhong Z,
Chen Y, Ren Q, Li Q, Kirkland ER, Zhang Y, Qi Y
TITLE A CRISPR-Cpf1 system for efficient genome editing and
transcriptional repression in plants.
JOURNAL Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18.
PUBMED 28211909
REFERENCE 2 (bases 1 to 6939)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6939
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 133..152
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 386..403
/label=L4440
/note="L4440 vector, forward primer"
terminator 562..589
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 681..767
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 816..838
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 830..847
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 831..847
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 863..962
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 987..1007
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1032..4712
/codon_start=1
/product="CRISPR-associated protein from Lachnospiraceae
bacterium ND2006 (Zetsche et al., 2015)"
/label=LbCpf1
/translation="SKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAED
YKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKEI
AKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFSE
EAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEGE
FFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVLS
DRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKNG
PAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQL
QEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKDL
LDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYSK
DKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNGN
YEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKLI
DFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLVE
EGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKKE
ELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKIN
TEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTDY
HSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSGF
KNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQN
GFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALDY
KNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQQ
GDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRNY
EAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTSV
KH"
CDS 4713..4760
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
terminator 4791..5043
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 5088..5211
/gene="mutant version of attR"
/label=attR5
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(5228..5247)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5229..5247)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5252..5268)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5252..5268)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 5699..6490
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 6583..232
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgcgagcca
541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
781 ccctactctc gcgttaacgc ttgcatggat gttttcccag tcacgacgtt gtaaaacgac
841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg
961 ctccgaattc gcccttcacc atggctccta agaagaagcg gaaggttggt attcacgggg
1021 tgcctgcggc ttcaaagctc gagaaattca ccaactgtta ttcgttgagc aaaacactgc
1081 ggtttaaagc gattccagtc ggcaagactc aagagaatat agacaataag cggctgttgg
1141 tggaagatga aaagcgcgcg gaagactaca aaggggtgaa gaagttgttg gacagatact
1201 acctctcttt tatcaatgat gtcttgcact caatcaaatt gaagaatctg aacaactaca
1261 tctccctctt cagaaagaaa acaaggacag aaaaggagaa taaggaactt gaaaatttgg
1321 agatcaatct gaggaaagag atcgcgaaag cctttaaagg caacgaagga tacaaaagtc
1381 tgttcaagaa ggatataatt gagacaattt tgccagagtt cctcgatgac aaggacgaga
1441 ttgcgctggt caattcgttc aacggattca caacagcatt cacaggcttc tttgataatc
1501 gggaaaatat gttctctgag gaggcaaagt ccacttctat tgcgttcagg tgtatcaatg
1561 agaatctcac taggtacatt tccaacatgg atatctttga gaaggttgac gcaatttttg
1621 acaagcacga agttcaggag attaaggaga agatcctcaa ttccgattat gacgttgagg
1681 acttcttcga aggtgagttt tttaatttcg tgctcactca agagggtatc gacgtgtata
1741 atgcgatcat cggtgggttc gtgactgagt ccggtgaaaa gattaaggga ttgaacgagt
1801 atatcaacct ttacaaccaa aagacgaaac agaagctgcc aaagttcaag cctctttaca
1861 aacaggttct ttcagaccgc gagtcactct cgttctatgg ggagggctac acttcggatg
1921 aggaagtcct ggaggtgttc aggaatactc tcaataagaa ttcggagatt ttctcttcta
1981 taaaaaaact ggaaaagttg tttaagaatt ttgacgaata ctctagcgcc ggcatatttg
2041 tgaaaaacgg cccggccata tcaacgataa gtaaagatat cttcggcgaa tggaacgtga
2101 tcagagacaa atggaacgcg gagtatgacg atattcacct gaagaagaag gctgtcgtaa
2161 cggagaagta cgaggatgat cgcaggaaaa gcttcaaaaa gatcggaagt ttcagcctgg
2221 aacagttgca ggagtatgct gacgccgatc ttagcgtcgt cgagaagttg aaggagataa
2281 tcatccaaaa ggtcgacgag atatataaag tctatggatc aagtgaaaaa ctgttcgacg
2341 ccgacttcgt tttggagaag tccctgaaga agaacgacgc tgttgttgcc attatgaagg
2401 atctgctcga cagcgtgaag agtttcgaga actatattaa ggcttttttc ggggagggga
2461 aggagactaa cagagatgag tccttctacg gagacttcgt cctcgcgtac gatatactcc
2521 ttaaggtaga ccacatctac gacgcaatca gaaattacgt gacacaaaag ccgtacagca
2581 aggacaagtt caaactctac ttccagaacc cccagttcat gggcggctgg gacaaggaca
2641 aggaaacgga ttacagggct acgatcctga ggtatggttc aaaatactac ttggcgatta
2701 tggacaagaa gtacgccaag tgtctccaga agattgacaa agacgatgtc aatggcaatt
2761 atgagaagat caactacaag ctgcttccgg gtccgaacaa gatgctccca aaggttttct
2821 tcagcaagaa atggatggcc tactataacc caagcgagga catccagaag atttataaga
2881 acggtacgtt caagaagggc gacatgttca atcttaacga ctgtcacaag ctgatcgact
2941 tcttcaaaga ctcaattagc cggtacccaa agtggtctaa cgcctatgac ttcaactttt
3001 cggaaaccga gaagtacaag gatatagccg gattttatag agaggtggaa gagcagggct
3061 acaaggtgtc attcgagtcc gccagcaaga aggaagtgga caagctcgtg gaagagggta
3121 agctctacat gttccagatt tataataaag actttagcga taagagccac gggacaccta
3181 atctccacac aatgtatttc aagctgctct tcgacgagaa taaccacggc caaatcaggt
3241 tgtcaggagg ggctgaactc ttcatgcggc gcgctagcct taagaaggag gagcttgtag
3301 tccaccctgc gaatagtcca attgcgaata agaacccgga caatcctaaa aagactacaa
3361 cattgagcta cgacgtgtac aaggataaga ggttttccga ggatcagtac gagctccaca
3421 tcccgattgc gatcaacaag tgcccaaaga atattttcaa gataaacaca gaggtgcgtg
3481 tactcctgaa gcatgacgac aatccttacg tcattgggat tgatcggggc gagaggaacc
3541 tcctctatat tgtggtggtg gacgggaagg ggaacatagt cgaacagtac tcccttaacg
3601 aaataattaa caatttcaac ggcatccgta tcaagaccga ctaccattcg ttgctggaca
3661 agaaggagaa ggagagattt gaggcgcggc aaaattggac aagtatcgag aacatcaagg
3721 aactcaaagc aggttatatc tctcaagttg tgcataagat atgcgagctg gttgagaagt
3781 atgacgcagt gatcgctctt gaggacctca actcgggctt taagaattct agagttaaag
3841 tggagaagca ggtctatcaa aagttcgaga agatgcttat agataagctc aactacatgg
3901 tcgataagaa atcgaaccca tgtgccaccg gcggcgcact caaaggttac caaataacaa
3961 acaaattcga gtccttcaaa tcgatgagta ctcagaatgg gttcatattt tatataccgg
4021 cgtggcttac gtctaagatc gacccgtcaa ctggttttgt caacctgttg aagacgaaat
4081 acacgtccat tgccgattcg aaaaagttca tatctagttt tgatcgtatt atgtacgtcc
4141 cagaggaaga tcttttcgag tttgctctcg actacaaaaa cttttcgcgg accgatgcgg
4201 attacattaa aaaatggaaa ctctattcgt acggcaacag aatcaggatt tttcgcaacc
4261 ctaagaagaa taacgtcttt gattgggagg aagtttgctt gactagcgcg tacaaggagc
4321 tctttaataa gtatggcatt aactaccaac agggtgatat cagagcactg ctttgcgaac
4381 aatctgacaa ggctttctac tcatccttca tggctttgat gagcctgatg ctccagatga
4441 gaaattcaat tacaggcaga accgacgtgg atttcttgat ctccccggtt aaaaattctg
4501 atggcatctt ttacgatagc aggaactatg aagcgcaaga gaatgcgatt ctgccaaaaa
4561 atgcagacgc caacggtgcc tataacatcg ccaggaaagt cctgtgggcg atcggccagt
4621 tcaaaaaggc cgaagacgaa aaattggaca aggtcaaaat cgctatcagc aacaaagagt
4681 ggctggagta tgctcagaca tccgtaaagc ataagcgtcc tgctgccacc aaaaaggccg
4741 gacaggctaa gaaaaagaag tgagacgact agtggcggcc gccgacgtcc gatcgttcaa
4801 acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca
4861 tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat
4921 ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa
4981 acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag
5041 atcgggaatt gatcccccct cgacagcttc cggaaagggc gaattcgcaa ctttgtatac
5101 aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
5161 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat gccatccagc
5221 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
5281 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct
5341 ctggaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
5401 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
5461 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
5521 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
5581 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
5641 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
5701 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
5761 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
5821 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
5881 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
5941 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
6001 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
6061 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
6121 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
6181 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
6241 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
6301 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
6361 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
6421 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
6481 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
6541 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
6601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
6661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
6721 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
6781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
6841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
6901 aacggggggt tcgtgcacac agcccagctt ggagcgaac
//
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