| 质粒名称: | plRL2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 163631 |
| 存储实验室: | Jeremy Rock |
|
载体骨架基本信息 |
|
| 载体名称: | Addgene #115162 |
| 载体出品公司: | Rock Lab |
| 空载体大小: | 8631 bp |
| 完整质粒大小: | 8631 bp |
| 载体类型: | 细胞表达, CRISPR |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 启动子: | Tet-On |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | TTCCTGTGAAGAGCCATTGATAATG |
|
文献引用方法 |
|
| 材料和方法部分: |
plRL2 was a gift from Jeremy Rock (Addgene plasmid # 163631 ; http://n2t.net/addgene:163631 ; RRID:Addgene_163631)
|
| 参考文献部分: | Genome-wide gene expression tuning reveals diverse vulnerabilities of M. tuberculosis. Bosch B, DeJesus MA, Poulton NC, Zhang W, Engelhart CA, Zaveri A, Lavalette S, Ruecker N, Trujillo C, Wallach JB, Li S, Ehrt S, Chait BT, Schnappinger D, Rock JM. Cell. 2021 Jul 16. pii: S0092-8674(21)00824-2. doi: 10.1016/j.cell.2021.06.033. 10.1016/j.cell.2021.06.033 PubMed 34297925 |
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 plRL2 8631 bp ds-DNA circular SYN 10-AUG-2021
DEFINITION Optimized M. tuberculosis CRISPRi plasmid; L5 integrating..
ACCESSION .
VERSION .
KEYWORDS plRL2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8631)
AUTHORS Bosch B, DeJesus MA, Poulton NC, Zhang W, Engelhart CA, Zaveri A,
Lavalette S, Ruecker N, Trujillo C, Wallach JB, Li S, Ehrt S, Chait
BT, Schnappinger D, Rock JM
TITLE Genome-wide gene expression tuning reveals diverse vulnerabilities
of M. tuberculosis.
JOURNAL Cell. 2021 Jul 16. pii: S0092-8674(21)00824-2. doi:
10.1016/j.cell.2021.06.033.
PUBMED 34297925
REFERENCE 2 (bases 1 to 8631)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 10, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8631
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(496..1084)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(576..595)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
protein_bind 1267..1285
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1297..1315
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
terminator 4795..4822
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind complement(7588..7607)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 7599..8414
/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(7672..7691)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
ORIGIN
1 aataaacgaa aggctcagtc gaaagactgg gcctttcgtt tatctgttgt ttgaaaaaaa
61 aaagcgccgc aactgcggcg cttttttttt tgaattctct gaccagggaa aatagccctc
121 tgacctgggg atttgcgatc cctatcagtg atagatataa tctgggagga gacgattaat
181 gcgtctcggt ttttgtactc gaaagaagct acaaagataa ggcttcatgc cgaaatcaac
241 accctgtcat tttatggcag ggtgtttttt ttttgtcgac ttggggaccc tagaggtccc
301 cttttttatt tttttggatg aagagcatta tcaatggctc ttcacaggaa gcttaaaaaa
361 aaaagccccg cgattgcggg gctttttttt tttaatacgg ttatccacag aatcagggga
421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
661 tctcccctcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
901 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
1021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
1081 tcaagaagat cctttgatct tttctacggg gtctgacgct catggaacaa taaacgaaag
1141 gctcagtcga aagactgggc ctttcgttta tctgttgttt gaaaaaaaaa agccccgcga
1201 ttgcggggct tttttttttc atatgtctga ccagggaaaa tagccctctg acctggggat
1261 ttgacttccc tatcagtgat agagataatc tgggagtccc tatcagtgat agagaacgcg
1321 gtatcgatat gtcggacctg gtgctgggcc tggccatcgg catcggctcg gtgggcgtgg
1381 gcatcctgaa caaggtgacc ggcgagatca tccacaagaa ctcgcgcatc ttcccggccg
1441 cccaggccga gaacaacctg gtgcgccgca ccaaccgcca gggccgccgc ctggcccgcc
1501 gcaagaagca ccgccgcgtg cgcctgaacc gcctgttcga ggagtcgggc ctgatcaccg
1561 acttcaccaa gatctcgatc aacctgaacc cgtaccagct gcgcgtgaag ggcctgaccg
1621 acgagctgtc gaacgaggag ctgttcatcg ccctgaagaa catggtgaag caccgcggca
1681 tctcgtacct ggacgacgcc tcggacgacg gcaactcgtc ggtgggcgac tacgcccaga
1741 tcgtgaagga gaactcgaag cagctggaga ccaagacccc gggccagatc cagctggagc
1801 gctaccagac ctacggccag ctgcgcggcg acttcaccgt ggagaaggac ggcaagaagc
1861 accgcctgat caacgtgttc ccgacctcgg cctaccgctc ggaggccctg cgcatcctgc
1921 agacccagca ggagttcaac ccgcagatca ccgacgagtt catcaaccgc tacctggaga
1981 tcctgaccgg caagcgcaag tactaccacg gcccgggcaa cgagaagtcg cgcaccgact
2041 acggccgcta ccgcacctcg ggcgagaccc tggacaacat cttcggcatc ctgatcggca
2101 agtgcacctt ctacccggac gagttccgcg ccgccaaggc ctcgtacacc gcccaggagt
2161 tcaacctgct gaacgacctg aacaacctga ccgtgccgac cgagaccaag aagctgtcga
2221 aggagcagaa gaaccagatc atcaactacg tgaagaacga gaaggccatg ggcccggcca
2281 agctgttcaa gtacatcgcc aagctgctgt cgtgcgacgt ggccgacatc aagggctacc
2341 gcatcgacaa gtcgggcaag gccgagatcc acaccttcga ggcctaccgc aagatgaaga
2401 ccctggagac cctggacatc gagcagatgg accgcgagac cctggacaag ctggcctacg
2461 tgctgaccct gaacaccgag cgcgagggca tccaggaggc cctggagcac gagttcgccg
2521 acggctcgtt ctcgcagaag caggtggacg agctggtgca gttccgcaag gccaactcgt
2581 cgatcttcgg caagggctgg cacaacttct cggtgaagct gatgatggag ctgatcccgg
2641 agctgtacga gacctcggag gagcagatga ccatcctgac ccgcctgggc aagcagaaga
2701 ccacctcgtc gtcgaacaag accaagtaca tcgacgagaa gctgctgacc gaggagatct
2761 acaacccggt ggtggccaag tcggtgcgcc aggccatcaa gatcgtgaac gccgccatca
2821 aggagtacgg cgacttcgac aacatcgtga tcgagatggc ccgcgagacc aacgaggacg
2881 acgagaagaa ggccatccag aagatccaga aggccaacaa ggacgagaag gacgccgcca
2941 tgctgaaggc cgccaaccag tacaacggca aggccgagct gccgcactcg gtgttccacg
3001 gccacaagca gctggccacc aagatccgcc tgtggcacca gcagggcgag cgctgcctgt
3061 acaccggcaa gaccatctcg atccacgacc tgatcaacaa ctcgaaccag ttcgaggtgg
3121 acgccatcct gccgctgtcg atcaccttcg acgactcgct ggccaacaag gtgctggtgt
3181 acgccaccgc caaccaggag aagggccagc gcaccccgta ccaggccctg gactcgatgg
3241 acgacgcctg gtcgttccgc gagctgaagg ccttcgtgcg cgagtcgaag accctgtcga
3301 acaagaagaa ggagtacctg ctgaccgagg aggacatctc gaagttcgac gtgcgcaaga
3361 agttcatcga gcgcaacctg gtggacaccc gctacgcctc gcgcgtggtg ctgaacgccc
3421 tgcaggagca cttccgcgcc cacaagatcg acaccaaggt gtcggtggtg cgcggccagt
3481 tcacctcgca gctgcgccgc cactggggca tcgagaagac ccgcgacacc taccaccacc
3541 acgccgtgga cgccctgatc atcgccgcct cgtcgcagct gaacctgtgg aagaagcaga
3601 agaacaccct ggtgtcgtac tcggaggacc agctgctgga catcgagacc ggcgagctga
3661 tctcggacga cgagtacaag gagtcggtgt tcaaggcccc gtaccagcac ttcgtggaca
3721 ccctgaagtc gaaggagttc gaggactcga tcctgttctc gtaccaggtg gactcgaagt
3781 tcaaccgcaa gatctcggac gccaccatct acgccacccg ccaggccaag gtgggcaagg
3841 acaaggccga cgagacctac gtgctgggca agatcaagga catctacacc caggacggct
3901 acgacgcctt catgaagatc tacaagaagg acaagtcgaa gttcctgatg taccgccacg
3961 acccgcagac cttcgagaag gtgatcgagc cgatcctgga gaactacccg aacaagcaga
4021 tcaacgacaa gggcaaggag gtgccgtgca acccgttcct gaagtacaag gaggagcacg
4081 gctacatccg caagtactcg aagaagggca acggcccgga gatcaagtcg ctgaagtact
4141 acgactcgaa gctgggcaac cacatcgaca tcaccccgaa ggactcgaac aacaaggtgg
4201 tgctgcagtc ggtgtcgccg tggcgcgccg acgtgtactt caacaagacc accggcaagt
4261 acgagatcct gggcctgaag tacgccgacc tgcagttcga caagggcacc ggcacctaca
4321 agatctcgca ggagaagtac aacgacatca agaagaagga gggcgtggac tcggactcgg
4381 agttcaagtt caccctgtac aagaacgacc tgctgctggt gaaggacacc gagaccaagg
4441 agcagcagct gttccgcttc ctgtcgcgca ccatgccgaa gcagaagcac tacgtggagc
4501 tgaagccgta cgacaagcag aagttcgagg gcggcgaggc cctgatcaag gtgctgggca
4561 acgtggccaa ctcgggccag tgcaagaagg gcctgggcaa gtcgaacatc tcgatctaca
4621 aggtgcgcac cgacgtgctg ggcaaccagc acatcatcaa gaacgagggc gacaagccga
4681 agctggactt ctaaatactc tggccatttt tctttaccgc ctttgagtga gctgataccg
4741 ctcgccgcag ccgaacgacc gagcgcaacg cgtgcggccg ctgattagct aagcagaagg
4801 ccatcctgac ggatggcctt tttctagaaa aaaaaaaagc gccgcaactg cggcgctttt
4861 ttttttttaa ttaagtgata gattgcgatg cgtgtgctcg gttgatataa tctgtcagat
4921 caggaggact tcgcatgtcg cggttggata agtccaaggt catcaactcg gcgctggagc
4981 tgctgaacga ggtcggtatc gagggactga ccacgcgcaa gttggcccaa aagctcggcg
5041 tggaacaacc gaccctctac tggcacgtga agaacaagcg cgcgctgctg gatgcgttgg
5101 ccatcgagat gctggatcgc catcacacgc acttttgccc cctggagggc gagagctggc
5161 aggattttct gcggaacaac gcgaagtcgt tccggtgcgc cctcctgagc catcgggatg
5221 gcgccaaagt ccacttgggc acccgcccga ccgagaagca atacgagacc ctggagaacc
5281 agctggcctt cctctgtcaa cagggttttt cgttggagaa cgccctgtat gccctgtccg
5341 ccgtgggaca cttcaccttg ggctgcgtgt tggaggatca ggaacatcag gtggcgaaag
5401 aggaacggga gacccccacg acggattcga tgcccccgtt gttgcgccag gcgatcgaac
5461 tgtttgacca ccagggcgcc gaacccgcct ttttgttcgg gctggagctg atcatctgcg
5521 ggctggaaaa gcaactgaag tgcgaaagcg gttcgtgata acgccgaaaa ccccgcttcg
5581 gcggggtttt gccgcggtct gcgttgtcgg gaagatttaa aataaaaaag gggacctcta
5641 gggtccccaa ttaaatagtt aacactagta ttaaaaaaaa aagccccgcg attgcggggc
5701 tttttttttt cgacggcgag ggttccgacc gctgcaactc ccggtgcaac cttgtcccgg
5761 tctattctct tcactgcacc agctccaatc tggtgtgaat gcccctcgtc tgttcgcgca
5821 ggcggggggc tctattcgtt tgtcagcatc gaaagtagcc agatcaggga tgcgttgcaa
5881 ccgcgtatgc ccaggtcaga agagtcgcac aagagttgca gacccctgga aagaaaaatg
5941 gccagagggc gaaaacaccc tctgaccagc ggagcgggcg acgggaatcg aacccgcgta
6001 gctagtttgg aagaatgggt gtctgccgac cacatatatg ggccggtcaa gataggtttt
6061 taccccctct cggctgcatc ctctaagtgg aaagaaattg caggtcgtag aagcgcgttg
6121 aagcctgaga gttgcacagg agttgcaacc cggtagcctt gttcacgacg agaggagacc
6181 tagttggcac gtcgcggatg gggatcgctg aagactcagc gcagcgggag gatccaagcc
6241 tcatacgtca acccgcagga cggtgtgagg tactacgcgc tgcagaccta cgacaacaag
6301 atggacgccg aagcctggct cgcgggcgag aagcggctca tcgagatgga gacctggacc
6361 cctccacagg accgggcgaa gaaggcagcc gccagcgcca tcacgctgga ggagtacacc
6421 cggaagtggc tcgtggagcg cgacctcgca gacggcacca gggatctgta cagcgggcac
6481 gcggagcgcc gcatctaccc ggtgctaggt gaagtggcgg tcacagagat gacgccagct
6541 ctggtgcgtg cgtggtgggc cgggatgggt aggaagcacc cgactgcccg ccggcatgcc
6601 tacaacgtcc tccgggcggt gatgaacaca gcggtcgagg acaagctgat cgcagagaac
6661 ccgtgccgga tcgagcagaa ggcagccgat gagcgcgacg tagaggcgct gacgcctgag
6721 gagctggaca tcgtcgccgc tgagatcttc gtgcactacc ggatcgcggc atacatcctg
6781 gcgtggacga gcctccggtt cggagagctg atcgagcttc gccgcaagga catcgtggac
6841 gacggcatga cgatgaagct ccgggtgcgc cgtggcgctt cccgcgtggg gaacaagatc
6901 gtcgttggca acgccaagac cgtccggtcg aagcgtcctg tgacggttcc gcctcacgtc
6961 gcggagatga tccgagcgca catgaaggac cgtacgaaga tgaacaaggg ccccgaggca
7021 ttcctggtga ccacgacgca gggcaaccgg ctgtcgaagt ccgcgttcac caagtcgctg
7081 aagcgtggct acgccaagat cggtcggccg gaactccgca tccacgacct ccgcgctgtc
7141 ggcgctacgt tcgccgctca ggcaggtgcg acgaccaagg agctgatggc ccgcctcggt
7201 cacacgactc ctaggatggc gatgaagtac cagatggcgt ctgaggcccg cgacgaggct
7261 atcgctgagg cgatgtccaa gctggccaag acctcctgaa acgcaaaaag cccccctccc
7321 aaggacactg agtcctaaag aggggggttt cttgtcagta cgcgaagaac cacgcctggc
7381 cgcgagcgcc agcaccgccg ctctgtgcgg agacctgggc accagccccg ccgccgccag
7441 gagcattgcc gttcccgcca gctagccacc tgacgtcggg ggggggggaa agccacgttg
7501 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
7561 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
7621 cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
7681 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccc
7741 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
7801 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
7861 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
7921 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
7981 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
8041 gcctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
8101 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataat cttttgccat
8161 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
8221 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
8281 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
8341 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
8401 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
8461 cttgacggga cggcggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat
8521 cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa
8581 ctggtccacc tacaacaaag ctctcaccaa ccgtggctcc ctcacgatat c
//
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