| 质粒名称: | pYPQ284-RVRR |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 160327 |
| 存储实验室: | Yiping Qi |
|
载体骨架基本信息 |
|
| 载体名称: | pYPQ167 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 植物表达，CRISPR；网关兼容Cas12a入口克隆 |
| 筛选标记: | -- |
| 载体抗性: | 大观霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Mb2Cas12a RVRR variant |
| 物种来源: | Moraxella bovoculi AAX08_00205 |
| 插入大小: | -- |
| 基因突变: | N563R, K569V, N573R and K625R |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: |
NLS（插入件上的N端子）
NLS（插入件上的C端子） |
| 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: | pYPQ284-RVRR was a gift from Yiping Qi (Addgene plasmid # 160327 ; http://n2t.net/addgene:160327 ; RRID:Addgene_160327) |
| 参考文献部分: | Expanding the scope of plant genome engineering with Cas12a orthologs and highly multiplexable editing systems. Zhang Y, Ren Q, Tang X, Liu S, Malzahn AA, Zhou J, Wang J, Yin D, Pan C, Yuan M, Huang L, Yang H, Zhao Y, Fang Q, Zheng X, Tian L, Cheng Y, Le Y, McCoy B, Franklin L, Selengut JD, Mount SM, Que Q, Zhang Y, Qi Y. Nat Commun. 2021 Mar 29;12(1):1944. doi: 10.1038/s41467-021-22330-w. 10.1038/s41467-021-22330-w PubMed 33782402 |
买家导航
pYPQ284RVRR载体基本信息
pYPQ284RVRR载体图谱质粒图谱和多克隆位点信息
pYPQ284RVRR载体简介
pYPQ284RVRR载体序列
LOCUS pYPQ284-RVRR 7008 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gateway entry plasmid (attL1.
ACCESSION .
VERSION .
KEYWORDS pYPQ284-RVRR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7008)
AUTHORS Zhang Y, Ren Q, Tang X, Liu S, Malzahn AA, Zhou J, Wang J, Yin D,
Pan C, Yuan M, Huang L, Yang H, Zhao Y, Fang Q, Zheng X, Tian L,
Cheng Y, Le Y, McCoy B, Franklin L, Selengut JD, Mount SM, Que Q,
Zhang Y, Qi Y
TITLE Expanding the scope of plant genome engineering with Cas12a
orthologs and highly multiplexable editing systems.
JOURNAL Nat Commun. 2021 Mar 29;12(1):1944. doi: 10.1038/s41467-021-22330-w.
PUBMED 33782402
REFERENCE 2 (bases 1 to 7008)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7008
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 358..1149
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPARYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 1242..1830
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1731..1750
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1984..2001
/label=L4440
/note="L4440 vector, forward primer"
terminator 2160..2187
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2279..2365
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 2414..2436
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2428..2445
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2429..2445
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2461..2560
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2585..2605
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6380..6427
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
terminator 6458..6710
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 6755..6878
/gene="mutant version of attR"
/label=attR5
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(6895..6914)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6896..6914)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6919..6935)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6919..6935)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 tggaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
61 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
121 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
181 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
241 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
301 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
361 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
421 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
481 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
541 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
601 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
661 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
721 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
781 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
841 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
901 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
961 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccggtatcag
1021 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
1081 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
1141 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
1381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
2101 agtgagcgca acgcaattaa tacgcgtacc gcgagccagg aagagtttgt agaaacgcaa
2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgctt
2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgaattcgc ccttcaccat
2581 ggctcctaag aagaagcgga aggttggtat tcacggggtg cctgcggctc tgtttcaaga
2641 ttttacacat ctgtacccgc tgagtaaaac agtgcggttc gagctgaaac ccataggaag
2701 gaccctcgag cacatccacg cgaagaattt tctgagccag gatgaaacta tggctgatat
2761 gtatcaaaaa gttaaggtca ttttggacga ctatcatcgc gattttattg ccgacatgat
2821 gggagaggtg aaactcacga agcttgctga attttacgac gtctatctga agttcaggaa
2881 aaatcctaag gacgatgggc tgcaaaaaca gcttaaagac cttcaagctg tccttcggaa
2941 ggaatcggtg aagcctatag ggtcaggtgg gaagtacaaa acaggctacg atagactctt
3001 tggggcaaaa ctcttcaaag atggaaaaga gttgggtgac ctcgcaaaat tcgttatagc
3061 ccaagaaggt gagtcttctc cgaagctggc tcatcttgct cattttgaga agttcagcac
3121 gtattttact ggatttcacg ataatcggaa gaatatgtac tcggatgaag acaagcatac
3181 tgcaatagcg tacaggctca tccatgagaa tttgccgaga ttcatcgaca atctgcaaat
3241 cttgacaaca atcaaacaaa agcatagcgc cctctatgat cagataatca acgagctcac
3301 ggcctccggg ctcgacgtct ccttggcttc tcatcttgac gggtatcaca agctccttac
3361 acaagagggg atcacggcat acaacaggat cataggagag gtgaatggat atacaaataa
3421 gcataaccag atatgccaca agagcgagcg catagcgaaa cttagaccct tgcacaagca
3481 aatcctttct gacggaatgg gagtgtcatt ccttccgtct aagttcgcgg atgatagtga
3541 gatgtgccaa gcggtcaacg aattttatcg ccattatact gacgtgttcg caaaggtgca
3601 aagtctcttt gacggatttg atgatcacca gaaagacggg atctatgttg aacacaaaaa
3661 ccttaatgaa ctgagcaaac aggcgttcgg cgactttgct ttgctgggga gggtccttga
3721 tggatactac gtggacgttg tcaatccgga gttcaatgag cggttcgcaa aggccaagac
3781 tgacaatgcg aaagccaagc ttacaaaaga aaaggacaaa ttcattaaag gagtccactc
3841 actggcttcc ctcgaacaag caatagaaca ccatacagct agacacgacg atgagagtgt
3901 tcaagccgga aaacttggcc agtacttcaa acacggtttg gcgggggttg acaacccgat
3961 tcagaaaatt cacaataacc attcgacgat taaagggttt ctggaaaggg aaaggcctgc
4021 tggggaacgg gcgctcccga agatcaagtc aggaaaaaac ccagaaatga cacagctcag
4081 gcagctgaag gaacttttgg acaacgcatt gaatgtggcg cacttcgcta agctgctgac
4141 aactaaaaca accttggaca accaggatgg aaatttttac ggggagtttg gggtgcttta
4201 cgacgagctg gctaaaattc caactctcta caataaggtt agagattatc tctctcaaaa
4261 gcccttttct accgaaaagt ataagctcaa cttcggcaat ccgacccttc tccgcgggtg
4321 ggacctgaac gtggagaaag atcgctttgg ggttatactt cagaaggatg gatgctatta
4381 cttggcgctt cttgataagg ctcataaaaa agttttcgac aacgccccta acactggtaa
4441 gaacgtctac caaaagatgg tctacaaact gttgcccggc cccaacaaaa tgcttcctag
4501 ggtgtttttc gcaaaatcga atctcgacta ttataatcca tctgccgagc tccttgacaa
4561 atatgctaag gggacccata aaaagggtga taatttcaac ctgaaggact gccacgcgct
4621 tatcgacttt ttcaaagccg ggataaataa gcatccggag tggcaacatt ttggttttaa
4681 attttcgcca acgtcgtcct atcgcgacct ttccgatttc tatagggaag ttgaacctca
4741 ggggtaccag gtcaaatttg ttgacattaa tgcggactac attgatgaat tggtggagca
4801 agggaagctc tacctctttc aaatatataa caaagatttc tcgccaaaag cgcatggtaa
4861 accgaatctt cataccttgt actttaaagc acttttttca gaagataact tggcggaccc
4921 gatctacaag ctgaatgggg aagctcagat cttctacagg aaagcttcgt tggacatgaa
4981 cgagactacc atacatcgcg cgggagaggt gcttgagaac aaaaatcccg acaacccgaa
5041 aaagcggcaa ttcgtttacg acatcatcaa agacaaacgg tacacgcagg acaaatttat
5101 gctccacgtc cccattacca tgaattttgg agtccaaggc atgaccatta aggaattcaa
5161 caaaaaggtc aaccaaagta ttcagcaata cgatgaagtc aatgtcatag gcatagatcg
5221 gggagaaagg catctgttgt atcttaccgt gattaactct aagggtgaaa tactggagca
5281 acggtcactt aacgatataa ccacggcgtc cgcgaacggt acacaagtga ccactcccta
5341 ccacaaaata ttggataaaa gggagataga acgcttgaat gcccgcgttg gctggggtga
5401 gattgagacc atcaaagagc ttaaatcggg atatttgtct cacgtcgttc atcaaattaa
5461 ccaactcatg cttaagtaca atgcaatcgt tgtgctcgag gacctgaact ttggtttcaa
5521 aagagggagg ttcaaggtgg aaaaacaaat ttaccagaac tttgaaaacg cgcttatcaa
5581 gaaattgaat caccttgttt tgaaagataa ggcagatgac gaaatcgggt cgtataaaaa
5641 tgcactccag ttgacaaata atttcacgga tttgaagtcg atcggcaagc aaacagggtt
5701 cctcttttat gtgccagcgt ggaatacatc aaaaattgat ccggagacgg gatttgtcga
5761 cttgctgaag cctaggtatg agaacattgc ccaatctcag gcctttttcg gcaaattcga
5821 taaaatatgc tacaacacag acaaaggtta ttttgaattt cacattgatt acgccaaatt
5881 tacagataag gcgaaaaaca gcagacagaa atgggctatc tgttctcatg gggacaaacg
5941 ctatgtctac gataagacgg ctaatcaaaa taaaggcgcc gcaaaaggta ttaatgtgaa
6001 tgatgagctg aaaagcttgt ttgcccgcta ccatatcaat gataaacaac caaacttggt
6061 gatggacata tgccagaaca atgacaaaga attccacaag tcactcatgt gcctgcttaa
6121 aacccttttg gcgctgcggt atagcaatgc atctagcgat gaagacttta ttttgagtcc
6181 cgtggccaac gacgagggcg tgttttttaa ttcagccttg gcggacgata cgcagcccca
6241 gaatgcggac gcaaacggcg cgtaccacat tgcactgaag ggactgtggc ttctgaacga
6301 gctgaaaaat agcgacgacc tgaataaagt caagttggcc attgacaatc aaacctggtt
6361 gaatttcgct caaaatagaa agcgtcctgc tgccaccaaa aaggccggac aggctaagaa
6421 aaagaagtga gacgactagt ggcggccgcc gacgtccgat cgttcaaaca tttggcaata
6481 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
6541 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
6601 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
6661 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattgat
6721 cccccctcga cagcttccgg aaagggcgaa ttcgcaactt tgtatacaaa agttgaacga
6781 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
6841 acataatact gtaaaacaca acatatccag tcactatgcc atccagctga tatcccctat
6901 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
6961 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctc
//
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