| 质粒名称: | pJMP3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79875 |
| 存储实验室: | Carol Gross, Stanley Qi |
|
载体骨架基本信息 |
|
| 载体名称: | pDG1731 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6673bp |
| 完整质粒大小: | 7263 bp |
| 载体修饰: | -- |
| 载体类型: | -- |
| 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: | pJMP3 was a gift from Carol Gross & Stanley Qi (Addgene plasmid # 79875 ; http://n2t.net/addgene:79875 ; RRID:Addgene_79875) |
| 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 pJMP3 7263 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacillus subtilis sgRNA expression vector; integrates into thrC.
ACCESSION .
VERSION .
KEYWORDS pJMP3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7263)
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 7263)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7263
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA 858..933
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 954..983
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 999..1016
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1072..1089)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 1242..1288
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(5434..6022)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5514..5533)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6193..7053)
/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 6816..6835
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7054..7158)
/gene="bla"
/label=AmpR promoter
primer_bind 7226..7244
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 aattcatgta aaagatgagg ttggttcatt ctcgaaaatt acatctgtgt tctcagagcg
61 gggcgtgacg tttgaaaaaa tccttcagct gccaattaaa ggccatgatg agttagctga
121 aatcgtaatt gtcacacatc atacatcaga agctgatttc agtgatatcc tgcaaaacct
181 aaatgatttg gaagtcgttc aagaagtcaa aagcacatat cgtgtagaag ggaacggttg
241 gagctaatgt ggaaaggact tatccatcaa tataaagaat ttttacctgt aacagatcaa
301 acaccggcgc taactttaca tgaaggaaac acacctctta ttcacctgcc gaagctgtct
361 gagcagctcg gaattgagct tcatgtcaaa acggaaggcg tcaatcctac gggatcattt
421 aaagatcgcg gaatggttat ggctgtggca aaggcaaaag aagaaggcaa tgacacgatt
481 atgtgcgcgt caacaggtaa cacttccgct gctgcggcag catatgcagc ccgtgctaac
541 atgaaatgca ttgtcatcat cccgaacgga aaaattgcat ttggaaaact cgctcaagct
601 gtcatgtacg gagccgagat tatcgcaatt gacggaaact ttgacgatgc gcttaaaatt
661 gtccgttcca tctgtgagaa atcaccgatt gcccttgtca actcagtcaa cccttaccgc
721 attgaaggcc aaaaaactgc tgccttcgga tcctagaagc ttatcgaatt ccttattaac
781 gttgatataa tttaaatttt atttgacaaa aatgggctcg tgttgtacaa taaatgtaac
841 tttcagttta gcggtctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
901 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gaagcttggg cccgaacaaa
961 aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag
1021 tttaaacggt ctccagcttg gctgttttgg cggatgagag aagattttca gcctgataca
1081 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
1141 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
1201 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
1261 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaactgg atccttactc
1321 gagtctagac tgcaggcttc ctcgctcact gactcgaatt cctgcagccc tggcgaatgg
1381 cgattttcgt tcgtgaatac atgttataat aactataact aataacgtaa cgtgactggc
1441 aagagatatt tttaaaacaa tgaataggtt tacacttact ttagttttat ggaaatgaaa
1501 gatcatatca tatataatct agaataaaat taactaaaat aattattatc tagataaaaa
1561 atttagaagc caatgaaatc tataaataaa ctaaattaag tttatttaat taacaactat
1621 ggatataaaa taggtactaa tcaaaatagt gaggaggata tatttgaata catacgaaca
1681 aattaataaa gtgaaaaaaa tacttcggaa acatttaaaa aataacctta ttggtactta
1741 catgtttgga tcaggagttg agagtggact aaaaccaaat agtgatcttg actttttagt
1801 cgtcgtatct gaaccattga cagatcaaag taaagaaata cttatacaaa aaattagacc
1861 tatttcaaaa aaaataggag ataaaagcaa cttacgatat attgaattaa caattattat
1921 tcagcaagaa atggtaccgt ggaatcatcc tcccaaacaa gaatttattt atggagaatg
1981 gttacaagag ctttatgaac aaggatacat tcctcagaag gaattaaatt cagatttaac
2041 cataatgctt taccaagcaa aacgaaaaaa taaaagaata tacggaaatt atgacttaga
2101 ggaattacta cctgatattc cattttctga tgtgagaaga gccattatgg attcgtcaga
2161 ggaattaata gataattatc aggatgatga aaccaactct atattaactt tatgccgtat
2221 gattttaact atggacacgg gtaaaatcat accaaaagat attgcgggaa atgcagtggc
2281 tgaatcttct ccattagaac atagggagag aattttgtta gcagttcgta gttatcttgg
2341 agagaatatt gaatggacta atgaaaatgt aaatttaact ataaactatt taaataacag
2401 attaaaaaaa ttataaaaaa attgaaaaaa tggtggaaac acttttttca atttttttgt
2461 tttattattt aatatttggg aaatattcat tctaattggt aatcagattt tagaaaacaa
2521 taaacccttg cataggggga tctcgaggcc ttccgaaaat gcgcggcttt gaagctgagg
2581 gtgcggcggc aatcgtgcgc aatgaagtga ttgaaaatcc ggaaacaata gcgacagcca
2641 ttcgtatcgg aaacccggca agctgggaca aagctgtaaa ggcagccgag gaatccaatg
2701 ggaaaattga cgaagtcact gatgatgaaa tccttcacgc atatcagctg atcgcccgtg
2761 tagaaggcgt gtttgcagaa ccaggttctt gcgcgtctat cgcaggagtg ctgaaacagg
2821 tgaaatccgg agaaattccg aaaggcagca aggtcgtagc tgtgttaaca ggaaacggac
2881 tgaaagatcc gaacacagcg gtcgacattt cagaaatcaa gcctgtcaca ttgccgactg
2941 atgaagacag catccttgaa tatgtaaaag gagcggcccg tgtatgaacg aagccgacat
3001 gctgttctct gtcactgttc ccggaagcac agctaaccta ggccccggct ttgattcagt
3061 cggaatggcg ctcagcagat atttgaagct gaccgtcttt gaaagcgaca aatggtcttt
3121 tgaggctgaa acagaaacag tcgccggaat taattcgagc tcctttaact tcctcaccag
3181 tagttgtatc ggtaccataa gtagaagcag caacccaagt agctttacca gcatccggtt
3241 caaccagcat agtaagaatc ttactggaca tcggcagttc ttcgaacagt gcgccaacta
3301 ccagctcttt ctgcagttca ttcagggcac cggagaacct gcgtgcaatc catcttgttc
3361 aatcatgcga aacgatcctc atcctgtctc ttgatccatg gattacgcgt taacccgggc
3421 ccgcggatgc atatgatcag atcctttaac tctggcaacc ctcaaaattg aatgagacat
3481 gctacacctc cggataataa atatatataa acgtatatag atttcataaa gtctaacaca
3541 ctagacttat ttacttcgta attaagtcgt taaaccgtgt gctctacgac caaaactata
3601 aaacctttaa gaactttctt tttttacaag aaaaaagaaa ttagataaat ctctcatatc
3661 ttttattcaa taatcgcatc cgattgcagt ataaatttaa cgatcactca tcatgttcat
3721 atttatcaga gctcgtgcta taattatact aattttataa ggaggaaaaa atatgggcat
3781 ttttagtatt tttgtaatca gcacagttca ttatcaacca aacaaaaaat aagtggttat
3841 aatgaatcgt taataagcaa aattcatata accaaattaa agagggttat aatgaacgag
3901 aaaaatataa aacacagtca aaactttatt acttcaaaac ataatataga taaaataatg
3961 acaaatataa gattaaatga acatgataat atctttgaaa tcggctcagg aaaaggccat
4021 tttacccttg aattagtaaa gaggtgtaat ttcgtaactg ccattgaaat agaccataaa
4081 ttatgcaaaa ctacagaaaa taaacttgtt gatcacgata atttccaagt tttaaacaag
4141 gatatattgc agtttaaatt tcctaaaaac caatcctata aaatatatgg taatatacct
4201 tataacataa gtacggatat aatacgcaaa attgtttttg atagtatagc taatgagatt
4261 tatttaatcg tggaatacgg gtttgctaaa agattattaa atacaaaacg ctcattggca
4321 ttacttttaa tggcagaagt tgatatttct atattaagta tggttccaag agaatatttt
4381 catcctaaac ctaaagtgaa tagctcactt atcagattaa gtagaaaaaa atcaagaata
4441 tcacacaaag ataaacaaaa gtataattat ttcgttatga aatgggttaa caaagaatac
4501 aagaaaatat ttacaaaaaa tcaatttaac aattccttaa aacatgcagg aattgacgat
4561 ttaaacaata ttagctttga acaattctta tctcttttca atagctataa attatttaat
4621 aagtaagtta agggatgcat aaactgcatc ccttaacttg tttttcgtgt gcctattttt
4681 tgtgaatcga ttatgtcttt tgcgcagtcg gcttaaacca gttttcgctg gtgcgaaaaa
4741 agagtgtctt gtgacaccta aattcaaaat ctatcggtca gatttatacc gatttgattt
4801 tatatattct tgaataacat acgccgagtt atcacataaa agcgggaacc aatcatcaaa
4861 tttaaacttc attgcataat ccattaaact cttaaattct acgattcctt gttcatcaat
4921 aaactcaatc atttctttaa ttaatttata tctatctgtt gttgttttct ttaataattc
4981 atcaacatct acaccgccat aaactatcat atcttctttt tgatatttaa atttattagg
5041 atcttaaggc ctaggtctag agtctttgtt ttgacgccat tagcgtacgt aacaatcctc
5101 gttaaaggac aaggacctga gcggaagtgt atcgtacagt agacggagta tactagtata
5161 gtctatagtc cgtggaatta ttatatttat ctccgacgat attctcatca gtgaaatcca
5221 gctggagttc tttagcaaat ttttttatta gctgaactta gtattagtgg ccatactcct
5281 ccaatccaaa gctatttaga aagattacta tatcctcaaa caggcggtaa ccggcctctt
5341 catcgggaat gcgcgcgacc ttcagcatcg ccggcatgtc cccctggcgg acgggaagta
5401 tccagctcga ggtcgggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
5461 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
5521 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
5581 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
5641 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
5701 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
5761 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
5821 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
5881 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg atagctcttg
5941 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
6001 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
6061 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
6121 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
6181 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
6241 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
6301 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
6361 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
6421 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
6481 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
6541 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
6601 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
6661 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
6721 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
6781 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
6841 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
6901 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
6961 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
7021 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
7081 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
7141 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
7201 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
7261 att
//
