| 质粒名称: | pSBY1_FnCpf1cg |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104622 |
| 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: | 8307bp |
| 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: | 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: | pSBY1_FnCpf1cg was a gift from Sheng Yang (Addgene plasmid # 104622 ; http://n2t.net/addgene:104622 ; RRID:Addgene_104622) |
| 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 pSBY1_FnCpf1cg 8307 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CRISPR system used to genome editing in Mycobacterium smegmatis,
expresses Fn CPf1.
ACCESSION .
VERSION .
KEYWORDS pSBY1_FnCpf1cg
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8307)
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 8307)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8307
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(128..147)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 139..954
/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(212..231)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 1285..1873
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1774..1793
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 8239..8285
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 gctagccacc tgacgtcggg ggggggggaa agccacgttg tgtctcaaaa tctctgatgt
61 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
121 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc gaggccgcga
181 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
241 caatcaggtg cgacaatcta tcgcttgtat gggaagcccc atgcgccaga gttgtttctg
301 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
361 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
421 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct
481 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
541 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
601 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
661 gttgaacaag tctggaaaga aatgcataat cttttgccat tctcaccgga ttcagtcgtc
721 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
781 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
841 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
901 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
961 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcggcttt
1021 gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg
1081 cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag
1141 ctctcaccaa ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt
1201 atgagtcagc aacaccttct tcacgaggca gacctcacta gttccatgag cgtcagaccc
1261 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
1321 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
1381 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
1441 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
1501 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
1561 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
1621 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
1681 agaaagcgcc acgcttcccg aggggagaaa ggcggacagg tatccggtaa gcggcagggt
1741 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
1801 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
1861 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
1921 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
1981 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcaacg cgtgagccca
2041 ccagctccgt aagttcgggc gctgtgtggc tcgtacccgc gcattcaggc ggcagggggt
2101 ctaacgggtc taaggcggcg tgtacggccg ccacagcggc tctcagcggc ccggaaacgt
2161 cctcgaaacg acgcatgtgt tcctcctggt tggtacaggt ggttgggggt gctcggctgt
2221 cgctggtgtt ccaccaccag ggctcgacgg gagagcgggg gagtgtgcag ttgtggggtg
2281 gcccctcagc gaaatatctg acttggagct cgtgtcggac catacaccgg tgattaatcg
2341 tggtctacta ccaagcgtga gccacgtcgc cgacgaattt gagcagctct ggctgccgta
2401 ctggccgctg gcaagcgacg atctgctcga ggggatctac cgccaaagcc gcgcgtcggc
2461 cctaggccgc cggtacatcg aggcgaaccc aacagcgctg gcaaacctgc tggtcgtgga
2521 cgtagaccat ccagacgcag cgctccgagc gctcagcgcc cgggggtccc atccgctgcc
2581 caacgcgatc gtgggcaatc gcgccaacgg ccacgcacac gcagtgtggg cactcaacgc
2641 ccctgttcca cgcaccgaat acgcgcggcg taagccgctc gcatacatgg cggcgtgcgc
2701 cgaaggcctt cggcgcgccg tcgatggcga ccgcagttac tcaggcctca tgaccaaaaa
2761 ccccggccac atcgcctggg aaacggaatg gctccactca gatctctaca cactcagcca
2821 catcgaggcc gagctcggcg cgaacatgcc accgccgcgc tggcgtcagc agaccacgta
2881 caaagcggct ccgacgccgc tagggcggaa ttgcgcactg ttcgattccg tcaggttgtg
2941 ggcctatcgt cccgccctca tgcggatcta cctgccgacc cggaacgtgg acggactcgg
3001 ccgcgcgatc tatgccgagt gccacgcgcg aaacgccgaa tttccgtgca acgacgtgtg
3061 tcccggaccg ctaccggaca gcgaggtccg cgccatcgcc aacagcattt ggcgttggat
3121 cacaaccaag tcgcgcattt gggcggacgg gatcgtggtc tacgaggcca cactcagtgc
3181 gcgccagtcg gccatctcgc ggaagggcgc agcagcgcgc acggcggcga gcacagttgc
3241 gcggcgcgca aagtccgcgt cagccatgga ggcattgcta tgagcgacgg ctacagcgac
3301 ggctacaacc ggcagccgac tgtccgcaaa aagcggcgcg tgaccgccgc cgaaggcgct
3361 cgaatcaccg gactatccga acgccacgtc gtccggctcg tggcgcagga acgcagcgag
3421 tggctcgccg agcaggctgc acgccgcgaa cgcatccgcg cctatcacga cgacgagggc
3481 cactcttggc cgcaaacggc caaacatttc gggctgcatc tggacaccgt taagcgactc
3541 ggctatcggg cgaggaaaga gcgtgcggca gaacaggaag cggctcaaaa ggcccacaac
3601 gaagccgaca atccaccgct gttctaacgc aattggggag cgggtgtcgc gggggttccg
3661 tggggggttc cgttgcaacg ggtcggacag gtaaaagtcc tggtagacgc tagttttctg
3721 gtttgggcca tgcctgtctc gttgcgtgtt tcgttgcgcc cgttttgaat accagccaga
3781 cgagacgggg ttctacgaat cttggtcgat accaagccat ttccgctgaa tatcggggag
3841 ctcaccgcca gaatcggtgg ttgtggtgat gtacgtggcg aactccgttg tagtgcctgt
3901 ggtggcatcc gtggcgcggc cgcggtacca gatctttaaa tctagacggt gaccacaacg
3961 cgcccgcttt gatcggggac gtctgcggcc gaccatttac gggtcttgtt gtcgttggcg
4021 gtcatgggcc gaacatactc acccggatcg gagggccgag gacaaggtcg aacgaggggc
4081 atgacccggt gcggggcttc ttgcactcgg cataggcgag tgctaagaat aacgttggca
4141 ctcgcgaccg gtgagtgcta ggtcgggacg gtgaggccag gcccgtcgtc gcagcgagtg
4201 gcagcgagga caacttgagc cgtccgtcgc gggcactgcg cccggccagc gtaagtagcg
4261 gggttgccgt cacccggtga cccccgtttc atccccgatc cggaggaatc acttcgcaat
4321 gtccatctac caagagtttg tgaataaata ctccctgtcc aagaccctcc gttttgagct
4381 gatcccccaa ggcaagaccc tcgaaaacat caaggcacgc ggcctcatcc tggatgacga
4441 aaagcgcgct aaggattaca agaaggcaaa gcagatcatc gacaagtacc accagttctt
4501 catcgaagag atcctgtcct ccgtgtgcat ctccgaggac ctgctccaga actactccga
4561 tgtctacttc aagctcaaga agtccgatga cgataacctg cagaaggact tcaagtccgc
4621 taaggatacc atcaagaagc agatctccga atacatcaag gattccgaga agttcaagaa
4681 cctcttcaac cagaacctga tcgacgcaaa gaagggccag gaatccgatc tcatcctgtg
4741 gctcaagcag tccaaggata acggcatcga gctcttcaag gccaactccg acatcaccga
4801 catcgatgaa gctctggaga tcatcaagtc cttcaagggc tggaccacct acttcaaggg
4861 cttccacgaa aaccgcaaga acgtgtactc ctccaacgat atcccaacct ctatcatcta
4921 ccgcatcgtc gacgataacc tgccaaagtt cctcgaaaac aaggcaaagt acgagtccct
4981 gaaggataag gccccagaag ctatcaacta cgagcagatc aagaaggacc tggccgaaga
5041 gctcaccttc gacatcgatt acaagacctc tgaagtgaac cagcgcgtct tctccctcga
5101 tgaagtgttc gagatcgcca acttcaacaa ctacctgaac cagtccggca tcaccaagtt
5161 caacaccatc atcggcggca agttcgtcaa cggcgaaaac accaagcgca agggcatcaa
5221 cgagtacatc aacctctact cccagcagat caacgataag accctgaaga agtacaagat
5281 gtccgtgctc ttcaagcaga tcctgtccga caccgaatcc aagtccttcg tcatcgacaa
5341 gctggaggac gattccgatg tggtcaccac catgcagtcc ttctacgaac agatcgcagc
5401 cttcaagacc gtggaagaga agtccatcaa ggagaccctc tccctgctct tcgacgatct
5461 gaaggctcag aagctggatc tctccaagat ctacttcaag aacgacaagt ccctgaccga
5521 tctctcccag caggtcttcg acgattactc cgtgatcggc accgcagtcc tggaatacat
5581 cacccagcag atcgccccaa agaacctcga taacccatcc aagaaggaac aggagctgat
5641 cgccaagaag accgaaaagg ctaagtacct gtccctcgag accatcaagc tggctctcga
5701 agagttcaac aagcaccgcg acatcgataa gcagtgccgc ttcgaagaga tcctcgcaaa
5761 cttcgctgca atcccaatga tcttcgacga aatcgcacag aacaaggata acctggccca
5821 gatctccatc aagtaccaga accagggcaa gaaggatctg ctccaggcct ccgctgagga
5881 cgatgtgaag gcaatcaagg acctgctcga tcagaccaac aacctgctcc acaagctgaa
5941 gatcttccac atctcccagt ccgaagacaa ggccaacatc ctcgacaagg atgagcactt
6001 ctacctggtg ttcgaagagt gctacttcga actcgctaac atcgtcccac tgtacaacaa
6061 gatccgcaac tacatcaccc agaagccata ctccgatgaa aagttcaagc tcaacttcga
6121 gaactccacc ctggcaaacg gctgggacaa gaacaaggaa ccagataaca ccgccatcct
6181 cttcatcaag gacgataagt actacctggg cgtgatgaac aagaagaaca acaagatctt
6241 cgacgataag gccatcaagg aaaacaaggg cgagggctac aagaagatcg tgtacaagct
6301 gctcccaggc gctaacaaga tgctcccaaa ggtcttcttc tccgcaaagt ccatcaagtt
6361 ctacaaccca tccgaagata tcctgcgcat ccgcaaccac tccacccaca ccaagaacgg
6421 ctccccacag aagggctacg aaaagttcga gttcaacatc gaagactgcc gcaagttcat
6481 cgatttctac aagcagtcca tctccaagca cccagagtgg aaggacttcg gcttccgctt
6541 ctccgatacc cagcgctaca actccatcga tgaattctac cgcgaagtgg agaaccaggg
6601 ctacaagctg accttcgaaa acatctccga gtcctacatc gattccgtgg tcaaccaggg
6661 caagctgtac ctcttccaga tctacaacaa ggacttctcc gcttactcca agggccgccc
6721 aaacctgcac accctctact ggaaggcact cttcgacgaa cgcaacctgc aggatgtggt
6781 ctacaagctc aacggcgaag cagagctgtt ctaccgcaag cagtccatcc caaagaagat
6841 cacccaccca gccaaggaag caatcgccaa caagaacaag gataacccaa agaaggaatc
6901 cgtgttcgag tacgacctga tcaaggataa gcgcttcacc gaggacaagt tcttcttcca
6961 ctgcccaatc accatcaact tcaagtcctc cggcgccaac aagttcaacg atgaaatcaa
7021 cctgctcctg aaggagaagg ctaacgacgt gcacatcctg tccatcgatc gcggcgaacg
7081 ccacctcgcc tactacaccc tggtcgacgg caagggcaac atcatcaagc aggacacctt
7141 caacatcatc ggcaacgatc gcatgaagac caactaccac gacaagctgg ccgctatcga
7201 gaaggaccgc gattccgctc gcaaggattg gaagaagatc aacaacatca aggaaatgaa
7261 ggaaggctac ctctcccagg tggtccacga aatcgctaag ctggtgatcg agtacaacgc
7321 aatcgtggtc ttcgaagacc tgaacttcgg cttcaagcgc ggccgcttca aggtggagaa
7381 gcaggtctac cagaagctgg aaaagatgct catcgagaag ctgaactacc tcgtgttcaa
7441 ggacaacgaa ttcgataaga ccggcggcgt cctccgtgca taccagctga ccgccccatt
7501 cgagaccttc aagaagatgg gcaagcagac cggcatcatc tactacgtgc cagctggctt
7561 cacctctaag atctgcccag tgaccggctt cgtcaaccag ctctacccaa agtacgaatc
7621 cgtctccaag tcccaggagt tcttctccaa gttcgacaag atctgctaca acctggataa
7681 gggctacttc gaattctcct tcgactacaa gaacttcggc gataaggcag ccaagggcaa
7741 gtggaccatc gcatccttcg gctcccgcct catcaacttc cgcaactccg acaagaacca
7801 caactgggat acccgcgaag tgtacccaac caaggaactg gagaagctcc tgaaggatta
7861 ctccatcgaa tacggccacg gcgagtgcat caaggctgca atctgcggcg aatccgacaa
7921 gaagttcttc gcaaagctga cctctgtgct caacaccatc ctgcagatgc gcaactccaa
7981 gaccggcacc gagctggatt acctcatctc cccagtggcc gacgtcaacg gcaacttctt
8041 cgattcccgc caggctccaa agaacatgcc acaggacgct gatgcaaacg gcgcctacca
8101 catcggtctg aagggtctca tgctcctggg tcgcatcaag aacaaccagg aaggcaagaa
8161 gctgaatctc gtcattaaga acgaagaata ctttgaattt gtccagaacc gcaataacta
8221 agatcgactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
8281 tttatgccat catggccgcg gtgatca
//
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