| 质粒名称: | pSBY3_ligD |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104624 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pMV261 |
| 完整质粒大小: | 4989 bp |
| 载体类型: | Bacterial Expression, CRISPR |
| 载体抗性: | Kanamycin |
| 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: | FnCpf1 |
| 基因突变: | FnCpf1 codon-optimized C.glutamicum |
| 启动子: | hsp60 |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pSBY3_ligD was a gift from Sheng Yang (Addgene plasmid # 104624 ; http://n2t.net/addgene:104624 ; RRID:Addgene_104624) |
| 参考文献部分: | A CRISPR-Cpf1-Assisted Non-Homologous End Joining Genome Editing System of Mycobacterium smegmatis. Sun B, Yang J, Yang S, Ye RD, Chen D, Jiang Y. Biotechnol J. 2018 Sep;13(9):e1700588. doi: 10.1002/biot.201700588. Epub 2018 Aug 6. 10.1002/biot.201700588 PubMed 30039929 |
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 pSBY3_ligD 8617 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CRISPR system used to genome editing in Mycobacterium smegmatis,
expresses Fn Cpf1 and crRNA targeting ligD.
ACCESSION .
VERSION .
KEYWORDS pSBY3_ligD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8617)
AUTHORS Sun B, Yang J, Yang S, Ye RD, Chen D, Jiang Y
TITLE A CRISPR-Cpf1-Assisted Non-Homologous End Joining Genome Editing
System of Mycobacterium smegmatis.
JOURNAL Biotechnol J. 2018 Sep;13(9):e1700588. doi: 10.1002/biot.201700588.
Epub 2018 Aug 6.
PUBMED 30039929
REFERENCE 2 (bases 1 to 8617)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8617
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..53
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 145..230
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind complement(438..457)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 449..1264
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(522..541)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 1595..2183
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2084..2103
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 8549..8595
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 gctagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttaat
61 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc
121 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
181 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt aaatcaccgt
241 tgcgtgtgac gtgctatcta caacagtaga aattcggatc cattatacct aggactgagc
301 tagctgtcaa tctagccacc tgacgtcggg ggggggggaa agccacgttg tgtctcaaaa
361 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
421 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
481 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
541 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccc atgcgccaga
601 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
661 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
721 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
781 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
841 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
901 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
961 tggctggcct gttgaacaag tctggaaaga aatgcataat cttttgccat tctcaccgga
1021 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
1081 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
1141 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
1201 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
1261 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
1321 cggcggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
1381 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
1441 tacaacaaag ctctcaccaa ccgtggctcc ctcactttct ggctggatga tggggcgatt
1501 caggcctggt atgagtcagc aacaccttct tcacgaggca gacctcacta gttccatgag
1561 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
1621 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
1681 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
1741 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
1801 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
1861 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
1921 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
1981 gtgagctatg agaaagcgcc acgcttcccg aggggagaaa ggcggacagg tatccggtaa
2041 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
2101 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
2161 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
2221 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
2281 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcaacg
2341 cgtgagccca ccagctccgt aagttcgggc gctgtgtggc tcgtacccgc gcattcaggc
2401 ggcagggggt ctaacgggtc taaggcggcg tgtacggccg ccacagcggc tctcagcggc
2461 ccggaaacgt cctcgaaacg acgcatgtgt tcctcctggt tggtacaggt ggttgggggt
2521 gctcggctgt cgctggtgtt ccaccaccag ggctcgacgg gagagcgggg gagtgtgcag
2581 ttgtggggtg gcccctcagc gaaatatctg acttggagct cgtgtcggac catacaccgg
2641 tgattaatcg tggtctacta ccaagcgtga gccacgtcgc cgacgaattt gagcagctct
2701 ggctgccgta ctggccgctg gcaagcgacg atctgctcga ggggatctac cgccaaagcc
2761 gcgcgtcggc cctaggccgc cggtacatcg aggcgaaccc aacagcgctg gcaaacctgc
2821 tggtcgtgga cgtagaccat ccagacgcag cgctccgagc gctcagcgcc cgggggtccc
2881 atccgctgcc caacgcgatc gtgggcaatc gcgccaacgg ccacgcacac gcagtgtggg
2941 cactcaacgc ccctgttcca cgcaccgaat acgcgcggcg taagccgctc gcatacatgg
3001 cggcgtgcgc cgaaggcctt cggcgcgccg tcgatggcga ccgcagttac tcaggcctca
3061 tgaccaaaaa ccccggccac atcgcctggg aaacggaatg gctccactca gatctctaca
3121 cactcagcca catcgaggcc gagctcggcg cgaacatgcc accgccgcgc tggcgtcagc
3181 agaccacgta caaagcggct ccgacgccgc tagggcggaa ttgcgcactg ttcgattccg
3241 tcaggttgtg ggcctatcgt cccgccctca tgcggatcta cctgccgacc cggaacgtgg
3301 acggactcgg ccgcgcgatc tatgccgagt gccacgcgcg aaacgccgaa tttccgtgca
3361 acgacgtgtg tcccggaccg ctaccggaca gcgaggtccg cgccatcgcc aacagcattt
3421 ggcgttggat cacaaccaag tcgcgcattt gggcggacgg gatcgtggtc tacgaggcca
3481 cactcagtgc gcgccagtcg gccatctcgc ggaagggcgc agcagcgcgc acggcggcga
3541 gcacagttgc gcggcgcgca aagtccgcgt cagccatgga ggcattgcta tgagcgacgg
3601 ctacagcgac ggctacaacc ggcagccgac tgtccgcaaa aagcggcgcg tgaccgccgc
3661 cgaaggcgct cgaatcaccg gactatccga acgccacgtc gtccggctcg tggcgcagga
3721 acgcagcgag tggctcgccg agcaggctgc acgccgcgaa cgcatccgcg cctatcacga
3781 cgacgagggc cactcttggc cgcaaacggc caaacatttc gggctgcatc tggacaccgt
3841 taagcgactc ggctatcggg cgaggaaaga gcgtgcggca gaacaggaag cggctcaaaa
3901 ggcccacaac gaagccgaca atccaccgct gttctaacgc aattggggag cgggtgtcgc
3961 gggggttccg tggggggttc cgttgcaacg ggtcggacag gtaaaagtcc tggtagacgc
4021 tagttttctg gtttgggcca tgcctgtctc gttgcgtgtt tcgttgcgcc cgttttgaat
4081 accagccaga cgagacgggg ttctacgaat cttggtcgat accaagccat ttccgctgaa
4141 tatcggggag ctcaccgcca gaatcggtgg ttgtggtgat gtacgtggcg aactccgttg
4201 tagtgcctgt ggtggcatcc gtggcgcggc cgcggtacca gatctttaaa tctagacggt
4261 gaccacaacg cgcccgcttt gatcggggac gtctgcggcc gaccatttac gggtcttgtt
4321 gtcgttggcg gtcatgggcc gaacatactc acccggatcg gagggccgag gacaaggtcg
4381 aacgaggggc atgacccggt gcggggcttc ttgcactcgg cataggcgag tgctaagaat
4441 aacgttggca ctcgcgaccg gtgagtgcta ggtcgggacg gtgaggccag gcccgtcgtc
4501 gcagcgagtg gcagcgagga caacttgagc cgtccgtcgc gggcactgcg cccggccagc
4561 gtaagtagcg gggttgccgt cacccggtga cccccgtttc atccccgatc cggaggaatc
4621 acttcgcaat gtccatctac caagagtttg tgaataaata ctccctgtcc aagaccctcc
4681 gttttgagct gatcccccaa ggcaagaccc tcgaaaacat caaggcacgc ggcctcatcc
4741 tggatgacga aaagcgcgct aaggattaca agaaggcaaa gcagatcatc gacaagtacc
4801 accagttctt catcgaagag atcctgtcct ccgtgtgcat ctccgaggac ctgctccaga
4861 actactccga tgtctacttc aagctcaaga agtccgatga cgataacctg cagaaggact
4921 tcaagtccgc taaggatacc atcaagaagc agatctccga atacatcaag gattccgaga
4981 agttcaagaa cctcttcaac cagaacctga tcgacgcaaa gaagggccag gaatccgatc
5041 tcatcctgtg gctcaagcag tccaaggata acggcatcga gctcttcaag gccaactccg
5101 acatcaccga catcgatgaa gctctggaga tcatcaagtc cttcaagggc tggaccacct
5161 acttcaaggg cttccacgaa aaccgcaaga acgtgtactc ctccaacgat atcccaacct
5221 ctatcatcta ccgcatcgtc gacgataacc tgccaaagtt cctcgaaaac aaggcaaagt
5281 acgagtccct gaaggataag gccccagaag ctatcaacta cgagcagatc aagaaggacc
5341 tggccgaaga gctcaccttc gacatcgatt acaagacctc tgaagtgaac cagcgcgtct
5401 tctccctcga tgaagtgttc gagatcgcca acttcaacaa ctacctgaac cagtccggca
5461 tcaccaagtt caacaccatc atcggcggca agttcgtcaa cggcgaaaac accaagcgca
5521 agggcatcaa cgagtacatc aacctctact cccagcagat caacgataag accctgaaga
5581 agtacaagat gtccgtgctc ttcaagcaga tcctgtccga caccgaatcc aagtccttcg
5641 tcatcgacaa gctggaggac gattccgatg tggtcaccac catgcagtcc ttctacgaac
5701 agatcgcagc cttcaagacc gtggaagaga agtccatcaa ggagaccctc tccctgctct
5761 tcgacgatct gaaggctcag aagctggatc tctccaagat ctacttcaag aacgacaagt
5821 ccctgaccga tctctcccag caggtcttcg acgattactc cgtgatcggc accgcagtcc
5881 tggaatacat cacccagcag atcgccccaa agaacctcga taacccatcc aagaaggaac
5941 aggagctgat cgccaagaag accgaaaagg ctaagtacct gtccctcgag accatcaagc
6001 tggctctcga agagttcaac aagcaccgcg acatcgataa gcagtgccgc ttcgaagaga
6061 tcctcgcaaa cttcgctgca atcccaatga tcttcgacga aatcgcacag aacaaggata
6121 acctggccca gatctccatc aagtaccaga accagggcaa gaaggatctg ctccaggcct
6181 ccgctgagga cgatgtgaag gcaatcaagg acctgctcga tcagaccaac aacctgctcc
6241 acaagctgaa gatcttccac atctcccagt ccgaagacaa ggccaacatc ctcgacaagg
6301 atgagcactt ctacctggtg ttcgaagagt gctacttcga actcgctaac atcgtcccac
6361 tgtacaacaa gatccgcaac tacatcaccc agaagccata ctccgatgaa aagttcaagc
6421 tcaacttcga gaactccacc ctggcaaacg gctgggacaa gaacaaggaa ccagataaca
6481 ccgccatcct cttcatcaag gacgataagt actacctggg cgtgatgaac aagaagaaca
6541 acaagatctt cgacgataag gccatcaagg aaaacaaggg cgagggctac aagaagatcg
6601 tgtacaagct gctcccaggc gctaacaaga tgctcccaaa ggtcttcttc tccgcaaagt
6661 ccatcaagtt ctacaaccca tccgaagata tcctgcgcat ccgcaaccac tccacccaca
6721 ccaagaacgg ctccccacag aagggctacg aaaagttcga gttcaacatc gaagactgcc
6781 gcaagttcat cgatttctac aagcagtcca tctccaagca cccagagtgg aaggacttcg
6841 gcttccgctt ctccgatacc cagcgctaca actccatcga tgaattctac cgcgaagtgg
6901 agaaccaggg ctacaagctg accttcgaaa acatctccga gtcctacatc gattccgtgg
6961 tcaaccaggg caagctgtac ctcttccaga tctacaacaa ggacttctcc gcttactcca
7021 agggccgccc aaacctgcac accctctact ggaaggcact cttcgacgaa cgcaacctgc
7081 aggatgtggt ctacaagctc aacggcgaag cagagctgtt ctaccgcaag cagtccatcc
7141 caaagaagat cacccaccca gccaaggaag caatcgccaa caagaacaag gataacccaa
7201 agaaggaatc cgtgttcgag tacgacctga tcaaggataa gcgcttcacc gaggacaagt
7261 tcttcttcca ctgcccaatc accatcaact tcaagtcctc cggcgccaac aagttcaacg
7321 atgaaatcaa cctgctcctg aaggagaagg ctaacgacgt gcacatcctg tccatcgatc
7381 gcggcgaacg ccacctcgcc tactacaccc tggtcgacgg caagggcaac atcatcaagc
7441 aggacacctt caacatcatc ggcaacgatc gcatgaagac caactaccac gacaagctgg
7501 ccgctatcga gaaggaccgc gattccgctc gcaaggattg gaagaagatc aacaacatca
7561 aggaaatgaa ggaaggctac ctctcccagg tggtccacga aatcgctaag ctggtgatcg
7621 agtacaacgc aatcgtggtc ttcgaagacc tgaacttcgg cttcaagcgc ggccgcttca
7681 aggtggagaa gcaggtctac cagaagctgg aaaagatgct catcgagaag ctgaactacc
7741 tcgtgttcaa ggacaacgaa ttcgataaga ccggcggcgt cctccgtgca taccagctga
7801 ccgccccatt cgagaccttc aagaagatgg gcaagcagac cggcatcatc tactacgtgc
7861 cagctggctt cacctctaag atctgcccag tgaccggctt cgtcaaccag ctctacccaa
7921 agtacgaatc cgtctccaag tcccaggagt tcttctccaa gttcgacaag atctgctaca
7981 acctggataa gggctacttc gaattctcct tcgactacaa gaacttcggc gataaggcag
8041 ccaagggcaa gtggaccatc gcatccttcg gctcccgcct catcaacttc cgcaactccg
8101 acaagaacca caactgggat acccgcgaag tgtacccaac caaggaactg gagaagctcc
8161 tgaaggatta ctccatcgaa tacggccacg gcgagtgcat caaggctgca atctgcggcg
8221 aatccgacaa gaagttcttc gcaaagctga cctctgtgct caacaccatc ctgcagatgc
8281 gcaactccaa gaccggcacc gagctggatt acctcatctc cccagtggcc gacgtcaacg
8341 gcaacttctt cgattcccgc caggctccaa agaacatgcc acaggacgct gatgcaaacg
8401 gcgcctacca catcggtctg aagggtctca tgctcctggg tcgcatcaag aacaaccagg
8461 aaggcaagaa gctgaatctc gtcattaaga acgaagaata ctttgaattt gtccagaacc
8521 gcaataacta agatcgactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
8581 ggcctttcgt tttatgccat catggccgcg gtgatca
//
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