| 质粒名称: | pRH2502 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 84380 |
| 存储实验室: | Robert Husson |
|
载体骨架基本信息 |
|
| 载体名称: | pTE-10M-0X |
| 载体出品公司: | Addgene 20318 |
| 载体抗性: | 潮霉素 |
| 空载体大小: | 6608 bp |
| 完整质粒大小: | 6638 bp |
| 载体修饰: | 在pTE-10M-0X的ClaI位点合成并克隆了Pmyc1tetO（2）序列和2个BbsI位点，得到pRH2521 |
| 载体类型: | 细菌表达，CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | sgRNA cloning site |
| 物种来源: | -- |
| 插入大小: | 30 bp |
| 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: | ClaI(未破坏) |
| 3' 克隆位点: | ClaI (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: | pRH2521 was a gift from Robert Husson (Addgene plasmid # 84380 ; http://n2t.net/addgene:84380 ; RRID:Addgene_84380) |
| 参考文献部分: | Investigating essential gene function in Mycobacterium tuberculosis using an efficient CRISPR interference system. Singh AK, Carette X, Potluri LP, Sharp JD, Xu R, Prisic S, Husson RN. Nucleic Acids Res. 2016 Jul 12. pii: gkw625. 10.1093/nar/gkw625 PubMed 27407107 |
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 pRH2521 6624 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of sgRNA from a imyc promoter (Pimyc).
ACCESSION .
VERSION .
KEYWORDS pRH2521
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6624)
AUTHORS Singh AK, Carette X, Potluri LP, Sharp JD, Xu R, Prisic S, Husson RN
TITLE Investigating essential gene function in Mycobacterium tuberculosis
using an efficient CRISPR interference system.
JOURNAL Nucleic Acids Res. 2016 Jul 12. pii: gkw625.
PUBMED 27407107
REFERENCE 2 (bases 1 to 6624)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6624
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 37..53
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 38..54
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 197..224
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 424..441
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_RNA 503..578
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 1588..1615
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind complement(1705..1722)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1876..2464)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1956..1975)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 aacgttctcg gctcgatgat ccccgggggc cccccctcga ggtcgacggt atcgataata
61 ttggatcgtc gcaccgggtt aagcctggag tgcggtggtg cctggtcggc attttcgcag
121 tcgagggctc tcgtgtagcc tgggcgagtt gccgacgcag gcgaccctcc tgccacggat
181 ccgtgattag ctaagcagaa ggctatcctg acggatggcc tttttgcgtt taatactgtt
241 taaactctag aaatattgga tcgtcggcac cgtcacggcc gtgggaggcg gcacgatccg
301 cgacgtgatg atcggccgca tccccacggt gctgcgcagt gagctctacg ccatcccggc
361 gttgatctgt gcgttcgcac gcacaggccc ggtgtgagaa gggtctctga cgagcgggag
421 aactccctat cagtgataga gtttgtcctc cctatcagtg atagataggc tatgggagtg
481 tcttcgagac agaggaagac atgttttaga gctagaaata gcaagttaaa ataaggctag
541 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttatcgat ttaattaata
601 tgcatcaatt gatttaaata ctagtgttaa ctatttaatt ggggacccta gaggtcccct
661 tttttatttt aaaaattttt tcaaggggac ctctagggtc cccagctggc tagcgagtca
721 tgaggtcacc atggtcatta ggagccgctc tcgcacttca gctgcttctc caagccgcag
781 atgatcagct ccaggccgaa caagaaggcc ggctcggcgc cctggtggtc gaacagctcg
841 atggcctggc gcagcaacgg cggcatgctg tcggtggtcg gggtctcgcg ctcctccttg
901 gcgacctggt gctcctggtc ctccaacacg cagcccaggg tgaagtggcc cacggccgac
961 agggcgtaca acgcgttctc cagcgagaag ccctgctggc acaggaaggc caactggttc
1021 tccagggtct cgtactgctt ctcggtcggc cgggtgccca ggtgcacctt ggcgccgtcg
1081 cggtggctca gcaaggcgca gcggaagctc ttggcgttgt tgcgcaggaa gtcctgccag
1141 ctctcgccct ccagcgggca gaagtgggtg tggtggcggt ccaacatctc gatggccagg
1201 gcgtcgagca gggcccgctt gttcttgacg tgccagtaca gggtcggctg ctcgacgccc
1261 agcttctggg ccagcttgcg ggtggtcagg ccctcgatgc cgacctcgtt cagcagctcc
1321 agggcggagt tgatgacctt ggacttgtcc aggcgggaca ttgcgaagtg attcctcctg
1381 atggtaccgg atccgtggca ggagggtcgc ctgcgtcggc aactcgccca ggctacacga
1441 gagccctcga ctgcgaaaat gccgaccagg caccaccgca ctccaggctt aacccggtgc
1501 gacgatccaa tattacgaga tctgatatcc tcgaggcatg ccacgtgcct gcaggtttaa
1561 acgcggccgc aaacagtatt aaacgcaaaa aggccatccg tcaggatggc cttctgctta
1621 gctaatcaac tagttctagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
1681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
1741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
1801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
1861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
1921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
1981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
2041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
2101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
2161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
2221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
2281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
2341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
2461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
2521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
2641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
2701 ctgactcccc gacgtggccg accagcccgt catcgtcaac gcctgatccg cggtgcggac
2761 aggccgtgtc gcgaccggcc gtgcggaatt aagccggccc gtaccctgtg aatagaggtc
2821 cgctgtgaca caagaatccc tgttacttct cgaccgtatt gattcggatg attcctacgc
2881 gagcctgcgg aacgaccagg aattctggga gccgctggcc cgccgagccc tggaggagct
2941 cgggctgccg gtgccgccgg tgctgcgggt gcccggcgag agcaccaacc ccgtactggt
3001 cggcgagccc ggcccggtga tcaagctgtt cggcgagcac tggtgcggtc cggagagcct
3061 cgcgtcggag tcggaggcgt acgcggtcct ggcggacgcc ccggtgccgg tgccccgcct
3121 cctcggccgc ggcgagctgc ggcccggcac cggagcctgg ccgtggccct acctggtgat
3181 gagccggatg accggcacca cctggcggtc cgcgatggac ggcacgaccg accggaacgc
3241 gctgctcgcc ctggcccgcg aactcggccg ggtgctcggc cggctgcaca gggtgccgct
3301 gaccgggaac accgtgctca ccacccattc cgagatcttc ccggaactgc tgcgggaacg
3361 ccgcgcggcg accgtcgagg accaccgcgg gtggggctac ctctcgcccc ggctgctgga
3421 ccgcctggag gactggctgc cggacgtgga cacgctgctg gccggccgcg aaccccggtt
3481 cgtccacggc gacctgcacg ggaccaacat cttcgtggac ctggccgcga ccgaggtcac
3541 cgggatcgtc gacttcaccg acgtctatgc gggagactcc cgctacagcc tggtgcaact
3601 gcatctcaac gccttccggg gcgaccgcga gatcctggcc gcgctgctcg acggggcgca
3661 gtggaagcgg accgaggact tcgcccgcga actgctcgcc ttcaccttcc tgcacgactt
3721 cgaggtgttc gaggagaccc cgctggatct ctccggcttc accgatccgg aggaactggc
3781 gcagttcctc tgggggccgc cggacaccgc ccccggcgcc tgacgccccg ggccgcccgg
3841 cgccgccccc ggcccccggc ggccggccgc ccggagcccc gcccgcgctc gggagccccg
3901 ggcccgcgcc gaagcccgct gctgcgagcc cggagcgggc cggccgacgg cggtgcgggc
3961 ccggcggcgg acgctcagca gcggcgggcg tgaaaggccc tggcatcctc gatcatctcc
4021 tccagggtgg tcggccgagc ttccatccca gctcggcaag gatcaacgag gacagtcgca
4081 cgacgaagtt cttctggatc gcgcccgtgc tggaagcact caacctcgaa gcgtgtggtt
4141 gcggagccat ctagcaacca cacgaaacat gcgcaacgaa ccgcgcaacg aacaacgcct
4201 agaactggcc ctagatgagc tgactcgtat cgttggtaaa cctagtttga ccagcatgtt
4261 ttaactacgt tcggtgagct gtcaacgggg cctgtaacgg cacaacgaac cgtgcaacga
4321 gagtggccac ggatgccacc acaggcacta caacggagtt cgccacgtac atcaccacaa
4381 ccaccgattc tggcggtgag ctccccgata ttcagcggaa atggcttggt atcgaccaag
4441 attcgtagaa ccccgtctcg tctggctggt attcaaaacg ggcgcaacga aacacgcaac
4501 gagacaggca tggcccaaac cagaaaacta gcgtctacca ggacttttac ctgtccgacc
4561 cgttgcaacg gaacccccca cggaaccccc gcgacacccg ctccccaatt gcgttagaac
4621 agcggtggat tgtcggcttc gttgtgggcc ttttgagccg cttcctgttc tgccgcacgc
4681 tctttcctcg cccgatagcc gagtcgctta acggtgtcca gatgcagccc gaaatgtttg
4741 gccgtttgcg gccaagagtg gccctcgtcg tcgtgatagg cgcggatgcg ttcgcggcgt
4801 gcagcctgct cggcgagcca ctcgctgcgt tcctgcgcca cgagccggac gacgtggcgt
4861 tcggatagtc cggtgattcg agcgccttcg gcggcggtca cgcgccgctt tttgcggaca
4921 gtcggctgcc ggttgtagcc gtcgctgtag ccgtcgctca tagcaatgcc tccatggctg
4981 acgcggactt tgcgcgccgc gcaactgtgc tcgccgccgt gcgcgctgct gcgcccttcc
5041 gcgagatggc cgactggcgc gcactgagtg tggcctcgta gaccacgatc ccgtccgccc
5101 aaatgcgcga cttggttgtg atccaacgcc aaatgctgtt ggcgatggcg cggacctcgc
5161 tgtccggtag cggtccggga cacacgtcgt tgcacgggaa ttcggcgttt cgcgcgtggc
5221 actcggcata gatcgcgcgg ccgagtccgt ccacgttccg ggtcggcagg tagatccgca
5281 tgagggcggg acgataggcc cacaacctga cggaatcgaa cagtgcgcaa ttccgcccta
5341 gcggcgtcgg agccgctttg tacgtggtct gctgacgcca gcgcggcggt ggcatgttcg
5401 cgccgagctc ggcctcgatg tggctgagtg tgtagagatc tgagtggagc cattccgttt
5461 cccaggcgat gtggccgggg tttttggtca tgaggcctga gtaactgcgg tcgccgtcga
5521 cggcgcgccg aaggccttcg gcgcacgccg ccatgtatgc gagcggctta cgccgcgcgt
5581 attcggtgcg tggaacaggg gcgttgagtg cccacactgc gtgtgcgtgg ccgttggcgc
5641 gattgcccac gatcgcgttg ggcagcggat gggacccccg ggcgctgagc gctcggagcg
5701 ctgcgtctgg atggtctacg tccacgacca gcaggtttgc cagcgctgtt gggttcgcct
5761 cgatgtaccg gcggcctagg gccgacgcgc ggctttggcg gtagatcccc tcgagcagat
5821 cgtcgcttgc cagcggccag tacggcagcc agagctgctc aaattcgtcg gcgacgtggc
5881 tcacgcttgg tagtagacca cgattaatca ccggtgtatg gtccgacacg agctccaagt
5941 cagatatttc gctgaggggc caccccacaa ctgcacactc ccccgctctc ccgtcgagcc
6001 ctggtggtgg aacaccagcg acagccgagc acccccaacc acctgtacca accaggagga
6061 acacatgcgt cgtttcgagg acgtttccgg gccgctgaga gccgctgtgg cggccgtaca
6121 cgccgcctta gacccgttag accccctgcc gcctgaatgc gcgggtacga gccacacagc
6181 gcccgaactt acggagctgg tgggctcacc tggctttatg gcgtacgaat cggctgtgtg
6241 cgacctgttg ggcgaggtga ggtacgcgct actcacgctg gcaagggcga cagagccgcc
6301 ccaccgagcc cgcacggccg cgcgcggtgt caacaaccgg gtgagtcgtg cacaccagca
6361 ggtgttcgag gcttggctcg aagtgcagga catcgtggcg aacgccgccc gatgagccgc
6421 gccttacgct ggctgccagc cgttcgcggg ctggttggtg cagcgcgtcg agcggttaga
6481 ggccctgcgg tgttccacca ccgcagggcc tcgccctttt taaggctgaa tttgcttgtc
6541 tccgaatcca actggcttgt ccaagggtgt atctacgctt aatccaaagt tcaaacgagg
6601 ggattacaca tgaccaactt cgat
//
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