| 质粒名称: | pJMP2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79874 |
| 存储实验室: | Carol Gross, Stanley Qi |
|
载体骨架基本信息 |
|
| 载体名称: | pDG1662 |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6982 bp |
| 完整质粒大小: | 7572 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 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: | sgRNA RR1 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pJMP2 was a gift from Carol Gross & Stanley Qi (Addgene plasmid # 79874 ; http://n2t.net/addgene:79874 ; RRID:Addgene_79874) |
| 参考文献部分: | A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria. Peters JM, Colavin A, Shi H, Czarny TL, Larson MH, Wong S, Hawkins JS, Lu CH, Koo BM, Marta E, Shiver AL, Whitehead EH, Weissman JS, Brown ED, Qi LS, Huang KC, Gross CA. Cell. 2016 Jun 2;165(6):1493-506. doi: 10.1016/j.cell.2016.05.003. Epub 2016 May 26. 10.1016/j.cell.2016.05.003 PubMed 27238023 |
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 pJMP2 7572 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacillus subtilis sgRNA expression vector; integrates into amyE.
ACCESSION .
VERSION .
KEYWORDS pJMP2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7572)
AUTHORS Peters JM, Colavin A, Shi H, Czarny TL, Larson MH, Wong S, Hawkins
JS, Lu CH, Koo BM, Marta E, Shiver AL, Whitehead EH, Weissman JS,
Brown ED, Qi LS, Huang KC, Gross CA
TITLE A Comprehensive, CRISPR-based Functional Analysis of Essential Genes
in Bacteria.
JOURNAL Cell. 2016 Jun 2;165(6):1493-506. doi: 10.1016/j.cell.2016.05.003.
Epub 2016 May 26.
PUBMED 27238023
REFERENCE 2 (bases 1 to 7572)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7572
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA 745..820
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 841..870
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 886..903
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(959..976)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 1129..1175
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind complement(2363..2382)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
rep_origin complement(5743..6331)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5823..5842)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6502..7362)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7125..7144
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7363..7467)
/gene="bla"
/label=AmpR promoter
primer_bind 7535..7553
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga
61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt
121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat
181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt
241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg
301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg
361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg
421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa
481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat
541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca
601 catggaaaca cacaaattaa aaactggtct gatcggatcc tagaagctta tcgaattcct
661 tattaacgtt gatataattt aaattttatt tgacaaaaat gggctcgtgt tgtacaataa
721 atgtaacttt cagtttagcg gtctgtttta gagctagaaa tagcaagtta aaataaggct
781 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgaa gcttgggccc
841 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat
901 cattgagttt aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc
961 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
1021 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
1081 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
1141 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaactggatc
1201 cttactcgag tctagactgc aggcttcctc gctcactgac tcgaattctc atgtttgaca
1261 gcttatcatc ggcaatagtt acccttatta tcaagataag aaagaaaagg atttttcgct
1321 acgctcaaat cctttaaaaa aacacaaaag accacatttt ttaatgtggt ctttattctt
1381 caactaaagc acccattagt tcaacaaacg aaaattggat aaagtgggat atttttaaaa
1441 tatatattta tgttacagta atattgactt ttaaaaaagg attgattcta atgaagaaag
1501 cagacaagta agcctcctaa attcacttta gataaaaatt taggaggcat atcaaatgaa
1561 ctttaataaa attgatttag acaattggaa gagaaaagag atatttaatc attatttgaa
1621 ccaacaaacg acttttagta taaccacaga aattgatatt agtgttttat accgaaacat
1681 aaaacaagaa ggatataaat tttaccctgc atttattttc ttagtgacaa gggtgataaa
1741 ctcaaataca gcttttagaa ctggttacaa tagcgacgga gagttaggtt attgggataa
1801 gttagagcca ctttatacaa tttttgatgg tgtatctaaa acattctctg gtatttggac
1861 tcctgtaaag aatgacttca aagagtttta tgatttatac ctttctgatg tagagaaata
1921 taatggttcg gggaaattgt ttcccaaaac acctatacct gaaaatgctt tttctctttc
1981 tattattcca tggacttcat ttactgggtt taacttaaat atcaataata atagtaatta
2041 ccttctaccc attattacag caggaaaatt cattaataaa ggtaattcaa tatatttacc
2101 gctatcttta caggtacatc attctgtttg tgatggttat catgcaggat tgtttatgaa
2161 ctctattcag gaattgtcag ataggcctaa tgactggctt ttataatatg agataatgcc
2221 gactgtactt tttacagtcg gttttctaat gtcactaacc tgccccgtta gttgaagaag
2281 gtttttatat tacagctcca gatcctctac gccggacgca tcgtggccgg catcaccggc
2341 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
2401 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
2461 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
2521 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
2581 catggatgag cgatgatgat atccgtttag gctgggcggt gatagcttct cgttcaggca
2641 gtacgcctct tttcttttcc agacctgagg gaggcggaaa tggtgtgagg ttcccgggga
2701 aaagccaaat aggcgatcgc gggagtgctt tatttgaaga tcaggctatc actgcggtca
2761 atagatttca caatgtgatg gctggacagc ctgaggaact ctcgaacccg aatggaaaca
2821 accagatatt tatgaatcag cgcggctcac atggcgttgt gctggcaaat gcaggttcat
2881 cctctgtctc tatcaatacg gcaacaaaat tgcctgatgg caggtatgac aataaagctg
2941 gagcgggttc atttcaagtg aacgatggta aactgacagg cacgatcaat gccaggtctg
3001 tagctgtgct ttatcctgat gatattgcaa aagcgcctca tgttttcctt gagaattaca
3061 aaacaggtgt aacacattct ttcaatgatc aactgacgat taccttgcgt gcagatgcga
3121 atacaacaaa agccgtttat caaatcaata atggaccaga cgacaggcgt ttaaggatgg
3181 agatcaattc acaatcggaa aaggagatcc aatttggcaa aacatacacc atcatgttaa
3241 aaggaacgaa cagtgatggt gtaacgagga ccgagaaata cagttttgtt aaaagagatc
3301 cagcgtcggc caaaaccatc ggctatcaaa atccgaatca ttggagccag gtaaatgctt
3361 atatctataa acatgatggg agccgagtaa ttgaattgac cggatcttgg cctggaaaac
3421 caatgactaa aaatgcagac ggaatttaca cgctgacgct gcctgcggac acggatacaa
3481 ccaacgcaaa agtgattttt aataatggca gcgcccaagt gcccggtcag aatcagcctg
3541 gctttgatta cgtgctaaat ggtttatata atgactcggg cttaagcggt tctcttcccc
3601 attgagggca aggctagacg ggacttaccg aaagaaacca tcaatgatgg tttctttttt
3661 gttcataaat cagacaaaac ttttctcttg caaaagtttg tgaagtgttg cacaatataa
3721 atgtgaaata cttcacaaac aaaaagacat caaagagaaa cataccctgc aaggatgctg
3781 atattgtctg catttgcgcc ggagcaaacc aaaaacctgg tgagacacgc cttgaattag
3841 tagaaaagaa cttgaagatt ttcaaaggca tcgttagtga agtcatggcg agcggatttg
3901 acggcatttt cttagtcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg
3961 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg
4021 accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg
4081 gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat
4141 ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga
4201 gccgggccac ctactgaagt ggatttcttt aagagctcct ttaacttcct caccagtagt
4261 tgtatcggta ccataagtag aagcagcaac ccaagtagct ttaccagcat ccggttcaac
4321 cagcatagta agaatcttac tggacatcgg cagttcttcg aacagtgcgc caactaccag
4381 ctctttctgc agttcattca gggcaccgga gaacctgcgt gcaatccatc ttgttcaatc
4441 atgcgaaacg atcctcatcc tgtctcttga tccatggatt acgcgttaac ccgggcccgc
4501 ggatgcatat gatcagatct taaggcctag gtctagagga tcgatctgta taataaagaa
4561 taattattaa tctgtagaca aattgtgaaa ggatgtactt aaacgctaac ggtcagcttt
4621 attgaacagt aatttaagta tatgtccaat ctagggtaag taaattgagt atcaatataa
4681 actttatatg aacataatca acgaggtgaa atcatgagca atttgattaa cggaaaaata
4741 ccaaatcaag cgattcaaac attaaaaatc gtaaaagatt tatttggaag ttcaatagtt
4801 ggagtatatc tatttggttc agcagtaaat ggtggtttac gcattaacag cgatgtagat
4861 gttctagtcg tcgtgaatca tagtttacct caattaactc gaaaaaaact aacagaaaga
4921 ctaatgacta tatcaggaaa gattggaaat acggattctg ttagaccact tgaagttacg
4981 gttataaata ggagtgaagt tgtcccttgg caatatcctc caaaaagaga atttatatac
5041 ggtgagtggc tcaggggtga atttgagaat ggacaaattc aggaaccaag ctatgatcct
5101 gatttggcta ttgttttagc acaagcaaga aagaatagta tttctctatt tggtcctgat
5161 tcttcaagta tacttgtctc cgtacctttg acagatattc gaagagcaat taaggattct
5221 ttgccagaac taattgaggg gataaaaggt gatgagcgta atgtaatttt aaccctagct
5281 cgaatgtggc aaacagtgac tactggtgaa attacctcga aagatgtcgc tgcagaatgg
5341 gctatacctc ttttacctaa agagcatgta actttactgg atatagctag aaaaggctat
5401 cggggagagt gtgatgataa gtgggaagga ctatattcaa aggtgaaagc actcgttaag
5461 tatatgaaaa attctataga aacttctctc aattaggcta attttattgc aataacaggt
5521 gcttactttt ctggagttct ttagcaaatt tttttattag ctgaacttag tattagtggc
5581 catactcctc caatccaaag ctatttagaa agattactat atcctcaaac aggcggtaac
5641 cggcctcttc atcgggaatg cgcgcgacct tcagcatcgc cggcatgtcc ccctggcgga
5701 cgggaagtat ccagctcgag gtcgggccgc gttgctggcg tttttccata ggctccgccc
5761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
5821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
5881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
5941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
6001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
6061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
6121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
6181 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttga
6241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
6301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
6361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
6421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
6481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
6541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
6601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
6661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
6721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
6781 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
6841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
6901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
6961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
7021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
7081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
7141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
7201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
7261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
7321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
7381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
7441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
7501 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
7561 tcttcaagaa tt
//
