| 质粒名称: | pAGT8163 (AATG_ttLbCas12a-i_TTCG) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 192453 |
| 存储实验室: | Tom Schreiber |
|
载体骨架基本信息 |
|
| 载体名称: | pAGM3946 |
| 载体出品公司: | Sylvestre Marillonnet |
| 空载体大小: | 2073 bp |
| 完整质粒大小: | 7053 bp |
| 载体类型: | CRISPR, Synthetic Biology ; MoClo compatible Level 0 module |
| 载体抗性: | Spectinomycin |
| 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: | temperature-tolerant LbCas12a with introns (BsaI-flanked Golden Gate module, MoClo L0) / ttLbCas12a-i |
| 物种来源: | A. thaliana (mustard weed); Synthetic; Lachnospiraceae bacterium ND2006 |
| 插入大小: | 4988bp |
| 基因突变: | temperature-tolerant mutation (D156R) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BsaI (AATG) (destroyed during cloning) |
| 3’克隆位点: | BsaI (TTCG) (destroyed during cloning) |
| 5’测序引物 | CCTGTCGGGTTTCGCCACCTC |
| 3’测序引物 | GCCGTTACCACCGCTGCGTTC |
|
文献引用方法 |
|
| 材料和方法部分: | pAGT8163 (AATG_ttLbCas12a-i_TTCG) was a gift from Tom Schreiber (Addgene plasmid # 192453 ; http://n2t.net/addgene:192453 ; RRID:Addgene_192453) |
| 参考文献部分: | Enhancing gene editing and gene targeting efficiencies in Arabidopsis thaliana by using an intron-containing version of ttLbCas12a. Patrick Schindele, Laura Merker, Tom Schreiber, Anja Prange, Alain Tissier, Holger Puchta. Plant Biotechnology Journal, 16 November 2022 10.1111/pbi.13964 |
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 pAGT8163 (AATG_ttLbCas12a-i_TTCG) 7053 bp ds-DNA circular SYN 06-DEC-2022
DEFINITION intronized ttLbCas12a for in planta expression .
ACCESSION .
VERSION .
KEYWORDS pAGT8163 (AATG_ttLbCas12a-i_TTCG)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7053)
AUTHORS Patrick Schindele, Laura Merker, Tom Schreiber, Anja Prange, Alain
Tissier, Holger Puchta
TITLE Enhancing gene editing and gene targeting efficiencies in
Arabidopsis thaliana by using an intron-containing version of
ttLbCas12a
JOURNAL Plant Biotechnology Journal, 16 November 2022
REFERENCE 2 (bases 1 to 7053)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 6, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7053
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 301..1092
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 1433..2021
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1922..1941
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 2080..2100
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7004..7051
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
ORIGIN
1 ttcgtgagac cacgaagtgc cgaggcacga acccagtgga cataagcctg ttcggttcgt
61 aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
121 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta
181 cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg
241 ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatc
301 atgggggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
361 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
421 ggcctgaagc cacacagcga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
481 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
541 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
601 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
661 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
721 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
781 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
841 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
901 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
961 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
1021 tcgcgcgcag atcagttgga agaatttgtc cattacgtga aaggcgagat caccaaggta
1081 gtcggcaaat aatgtctagc tagaaattcg ttcaagccga cgccgcttcg cggcgcggct
1141 taactcaagc gttagatgca ctaagcacat aattgctcac agccaaacta tcaggtcaag
1201 tctgctttta ttatttttaa gcgtgcataa taagccctac acaaattggg agatatatca
1261 tgctgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
1321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
1381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
1441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
1501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
1561 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
1621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
1681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
1741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
1801 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
1861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
1921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
1981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
2041 ttttacggtt cctgcactct gtggtctcaa atgctgcagc ctaagaagaa gagaaaggtt
2101 ggaggagtcg actcgagtgc ggccgccaca atgagcaagc tcgagaagtt taccaactgc
2161 tacagcctgt ctaagaccct gaggttcaag gctattcctg tgggtaagac ccaagagaat
2221 atcgacaaca agcggctgct ggttgaggat gagaagagag ctgaggatta caagggcgtg
2281 aagaagctgc tggatcggta ctacctgagc ttcatcaacg atgtgctgca cagcatcaag
2341 ctgaagaacc tgaacaacta catcagcctg ttccggaaga aaacccggac cgagaaagag
2401 aacaaagagc ttgagaacct cgagatcaac ctgcggaaag agatcgctaa ggctttcaag
2461 ggtaacgaag gtaaggattt ttatgatata ctatgcttat gtattttgta ctgaaagcat
2521 atcctgcttc attgggatat tactgaaagc atttaactac atgtaaactc acttgatgat
2581 caataaactt gattttgcag gttacaagag cctgttcaag aaggatatta tcgagactat
2641 cctgcctgag ttcctggacg ataaggatga gattgccctg gtgaacagct tcaacggttt
2701 cactactgcc ttcaccggtt tcttcagaaa ccgggaaaac atgttcagcg aagaggccaa
2761 gtctacctct atcgctttcc ggtgcattaa cgagaacttg acccggtaca tcagcaacat
2821 ggacatcttc gagaaggtgg acgccatctt cgataagcac gaggtgcaag aaatcaaaga
2881 gaagatcctg aactccgact acgacgtcga ggattttttt gagggcgagt tcttcaactt
2941 cgtgctcacc caagaaggta acattcctta gttacctttc ttttcttttt ccatcataag
3001 tttatagatt gtacatgctt tgagattttt ctttgcaaac aatctcaggt atcgatgtgt
3061 acaacgctat catcggtggt ttcgtgactg agagcggtga gaagattaag ggcctgaacg
3121 agtacattaa cctgtacaat caaaagacca agcagaagct gccgaagttc aagccgcttt
3181 acaagcaggt tctgagcgat cgtgagagcc tgtcttttta cggagaggga tacacctctg
3241 atgaagaggt tttggaggta atcttgaaat tggaactctt cttttgttgt ctaaacctat
3301 caatttcttt gcggaaattt atttgaagct gtagagttaa aattgagtct tttaaacttt
3361 tgtaggtgtt ccgtaacacc ctgaacaaga acagcgagat cttcagctcc atcaagaagc
3421 tggaaaagct gtttaagaac ttcgacgagt acagcagcgc tggcatcttc gttaagaacg
3481 gtcctgctat cagcaccatc agcaaggata ttttcggcga gtggaacgtg atccgggata
3541 agtggaatgc tgagtacgat gacatccacc tgaagaaaaa ggctgtggtg accgagaagt
3601 acgaggatga taggcggaag tccttcaaga agataggtaa gttgttactt atgattgttt
3661 tcctctctgc tacatgtatt ttgttgttca tttctgtaag atataagaat tgagttttcc
3721 tctgatgata ttattaggtt cctttagcct cgagcagctt caagagtatg ctgacgctga
3781 tctgtccgtg gtcgagaagc ttaaagagat catcatccag aaggtcgacg agatctacaa
3841 ggtgtacggc agctctgaga agcttttcga tgctgacttc gtgttggaga agtctctgaa
3901 gaagaacgac gccgttgtcg ctatcatgaa ggatctgctg gacagcgtga agtctttcga
3961 gaactatatc aaggccttct tcggcgaagg ttagtatcat atgaagaaat acctagtttc
4021 agttgatgaa tgctattttc tgacctcagt tgttctcttt tgagaattat ttcttttcta
4081 atttgcctga tttttctatt aattcattag gtaaagagac taatagggac gagtcattct
4141 acggcgattt cgtgctggct tacgacatcc ttcttaaggt ggaccacatc tacgacgcca
4201 tcagaaatta cgtgacccag aagccgtaca gcaaggacaa gttcaagttg tacttccaga
4261 atccgcagtt catgggcggc tgggacaaag acaaagagac agattacagg gctaccatcc
4321 tgcggtacgg ctctaagtac taccttgcca tcatggacaa gaaatacgcc aagtgcctgc
4381 aaaagatcga caaggatgat gtgaacggca actacgagaa gatcaactac aagctcctgc
4441 caggtaaatc ctggtccaca cttttacgat aaaaacacaa gattttaaac tatgaactga
4501 tcaataatca ttcctaaaag accacacttt tgttttgttt ctaaagtaat ttttactgtt
4561 ataacaggtc ctaacaagat gcttcctaag gtgttcttct caaagaaatg gatggcctac
4621 tacaacccga gcgaggacat ccagaaaatc tacaagaacg gcaccttcaa aaagggcgac
4681 atgttcaacc tgaacgactg ccacaagctg atcgatttct tcaaggacag catcagccgg
4741 tatccgaagt ggtctaacgc ttacgatttc aacttcagcg agactgagaa gtataaggat
4801 atcgccggct tctaccgtga ggttgaggaa cagggttaca aggttagctt cgagagcgcc
4861 agcaagaaag aggtggacaa gttggttgaa gaaggtaaga ggtcaaaagg tttccgcaat
4921 gatccctctt tttttgtttc tctagtttca agaatttggg tatatgacta acttctgagt
4981 gttccttgat gcatatttgt gatgagacaa atgtttgttc tatgttttag gtaagctgta
5041 catgttccaa atctataaca aggacttctc cgacaagtct cacggcactc ctaatctgca
5101 tacaatgtac ttcaagctgc tgttcgacga gaacaaccac ggtcagatta ggctttctgg
5161 tggtgctgag ctgttcatga gaagggcctc actgaagaaa gaagagttgg tcgttcaccc
5221 tgccaactct ccaatcgcta acaagaaccc tgacaacccg aaaaagacca ccaccttgtc
5281 ttacgacgtg tacaaggata agcggttcag cgaggatcag tacgagcttc acattccgat
5341 cgccatcaac aagtgcccga agaacatctt caagatcaat accgaggtgc gggtgctgct
5401 gaagcacgat gataatcctt acgtgatcgg catcgatagg ggcgagagaa accttcttta
5461 catcgtggtg gtggacggca agggcaatat cgttgagcag tactctctga acgagattat
5521 caacaatttc aacggcatcc ggatcaagac cgactaccac tctctgctgg ataagaaaga
5581 aaaagagcgg ttcgaggcca ggcagaactg gacttctatc gaaaacatca aagagctgaa
5641 ggccggctac atctctcagg tggtgcataa gatttgcgag ctggtggaaa agtacgacgc
5701 tgtgattgct ctcgaggatc tgaacagcgg cttcaagaac tcacgtgtga aggtaaagca
5761 actgtgtttt aatcaatttc ttgtcaggat atatggatta taacttaatt tttgagaaat
5821 ctgtagtatt tggcgtgaaa tgagtttgct ttttggtttc tcccgtgtta taggttgaga
5881 agcaggtcta ccaaaagttc gagaagatgc tcatcgacaa gctgaactac atggtggaca
5941 aaaagagcaa cccttgcgct accggtggtg ctcttaaggg ttaccagatc actaacaagt
6001 tcgagtcttt caagagcatg agcacccaga acggcttcat cttctacatc cctgcttggc
6061 tgaccagcaa gatcgatcct tctactggct tcgtcaacct gctcaagacc aagtacacca
6121 gcattgccga cagcaagaag ttcatcagct cattcgaccg gatcatgtac gtgccagaag
6181 aggatctttt cgagttcgcc ctcgattaca agaacttctc taggaccgac gccgactaca
6241 ttaagaagtg gaagctgtac tcctacggca accggattcg gatctttcgg aacccgaaga
6301 aaaacaacgt gttcgactgg gaagaggtaa agtttccaac tttcctttac catatcaaac
6361 tatagttcga aactttttat ttgatcaact tcaaggccac ccgatctttc tattcctgat
6421 taatttgtga tgaatccata ttgacttttg atggttacgc aggtgtgcct gacctctgcc
6481 tacaaagaac tgttcaacaa gtacggcatc aactaccagc agggtgatat tagggctctg
6541 ctttgcgagc agtctgacaa ggctttctac agctctttca tggccctgat gtctctgatg
6601 ctgcaaatga ggaactctat caccggtagg accgatgtgg acttccttat ctctccggtg
6661 aagaacagtg acgggatctt ctacgacagc cggaattatg aggctcaaga gaacgcaatc
6721 ctgccgaaga atgctgatgc taacggcgct tacaacattg ccagaaaggt gctgtgggct
6781 atcggccagt ttaagaaagc cgaagatgag aagttggaca aggtctgtct ttcctatttc
6841 atatgtttaa tcctaggaat ttgatcaatt gattgtatgt atgtcgatcc caagactttc
6901 ttgttcactt atatcttaac tctctctttg ctgtttcttg caggtgaaga tcgctatctc
6961 caacaaagag tggctcgagt acgctcagac tagcgttaag cataaaaggc cggcggccac
7021 gaaaaaggcc ggccaggcaa aaaagaaaaa ggg
//
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