质粒名称: | PLJR962 |
---|---|
出品公司: | addgene |
目录编号: | 115162 |
存储实验室: | Sarah Fortune |
载体骨架基本信息 |
|
载体名称: | custom design |
完整质粒大小: | 8881bp |
载体类型: | 未定义 |
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 |
|
插入基因名称1: | Sth1 sgRNA scaffold |
物种来源: | Streptcoccus thermophilus |
插入大小: | 101bp |
插入基因名称2: | Sth1 dCas9 / nuclease-dead dcas9 allele |
物种来源: | Streptcoccus thermophilus |
插入大小: | 3366bp |
插入基因名称3: | TetR / Tet repressor protein |
插入大小: | 624bp |
插入基因名称4: | L5 attP |
插入大小: | 43bp |
插入基因名称5: | L5 Int |
插入大小: | 1116bp |
插入基因名称6: | KanR / Kanamycin resistance protein |
插入大小: | 816bp |
文献引用方法 |
|
材料和方法部分: | PLJR962 was a gift from Sarah Fortune (Addgene plasmid # 115162 ; http://n2t.net/addgene:115162 ; RRID:Addgene_115162) |
参考文献部分: | Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. Rock JM, Hopkins FF, Chavez A, Diallo M, Chase MR, Gerrick ER, Pritchard JR, Church GM, Rubin EJ, Sassetti CM, Schnappinger D, Fortune SM. Nat Microbiol. 2017 Feb 6;2:16274. doi: 10.1038/nmicrobiol.2016.274. 10.1038/nmicrobiol.2016.274 PubMed 28165460 |
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 PLJR962 8881 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Sth1 dCas9 TetR and KanR L5 Int attP for M. smegmatis. ACCESSION . VERSION . KEYWORDS PLJR962 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8881) AUTHORS Rock JM, Hopkins FF, Chavez A, Diallo M, Chase MR, Gerrick ER, Pritchard JR, Church GM, Rubin EJ, Sassetti CM, Schnappinger D, Fortune SM TITLE Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. JOURNAL Nat Microbiol. 2017 Feb 6;2:16274. doi: 10.1038/nmicrobiol.2016.274. PUBMED 28165460 REFERENCE 2 (bases 1 to 8881) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8881 /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" CDS 5212..5835 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind complement(5285..5304) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(7838..7857) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 7849..8664 /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(7922..7941) /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 agagaaggcg 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 ttaaagaaat attggatcgt cggcaccgtc acggccgtgg gaggcggcac 4921 gatccgcgac gtgatgatcg gccgcatccc cacggtgctg cgcagtgagc tctacgccat 4981 cccggcgttg atctgtgcgt tcgcacgcac aggcccggtg tgagaagggt ctctgacgag 5041 cgggagaacc cacccggggt gggcgagttt gtcctgcgtg tgctcggtcg agtaggctct 5101 gggagtaccc gtgtgtacga ccagcacggc atacatcatt tcgacgccga gagattcgcc 5161 gcccgaaatg agcacgatcc gcatggtacc atcaggagga atcacttcgc aatgtctaga 5221 ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt cggaatcgaa 5281 ggtttaacaa cccgtaaact cgcccagaag cttggtgtag agcagcctac attgtattgg 5341 catgtaaaaa ataagcgggc cctgctcgac gccttagcca ttgagatgtt agataggcac 5401 catactcact tttgcccttt agaaggggaa agctggcaag attttttacg taataacgct 5461 aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca tttaggtaca 5521 cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt atgccaacaa 5581 ggtttttcac tagagaacgc gttatatgca ctcagcgctg tggggcattt taccttaggt 5641 tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac acctactact 5701 gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca aggtgcagag 5761 ccagccttct tattcggcct tgaattgatc atctgcggat tagaaaaaca acttaaatgt 5821 gaaagtgggt cttaatgacc atgggctagc gctttagcta attaatttaa aataaaaaag 5881 gggacctcta gggtccccaa ttaaatagtt aacactagta ttaaaaaaaa aagccccgcg 5941 attgcggggc tttttttttt cgacggcgag ggttccgacc gctgcaactc ccggtgcaac 6001 cttgtcccgg tctattctct tcactgcacc agctccaatc tggtgtgaat gcccctcgtc 6061 tgttcgcgca ggcggggggc tctattcgtt tgtcagcatc gaaagtagcc agatcaggga 6121 tgcgttgcaa ccgcgtatgc ccaggtcaga agagtcgcac aagagttgca gacccctgga 6181 aagaaaaatg gccagagggc gaaaacaccc tctgaccagc ggagcgggcg acgggaatcg 6241 aacccgcgta gctagtttgg aagaatgggt gtctgccgac cacatatatg ggccggtcaa 6301 gataggtttt taccccctct cggctgcatc ctctaagtgg aaagaaattg caggtcgtag 6361 aagcgcgttg aagcctgaga gttgcacagg agttgcaacc cggtagcctt gttcacgacg 6421 agaggagacc tagttggcac gtcgcggatg gggatcgctg aagactcagc gcagcgggag 6481 gatccaagcc tcatacgtca acccgcagga cggtgtgagg tactacgcgc tgcagaccta 6541 cgacaacaag atggacgccg aagcctggct cgcgggcgag aagcggctca tcgagatgga 6601 gacctggacc cctccacagg accgggcgaa gaaggcagcc gccagcgcca tcacgctgga 6661 ggagtacacc cggaagtggc tcgtggagcg cgacctcgca gacggcacca gggatctgta 6721 cagcgggcac gcggagcgcc gcatctaccc ggtgctaggt gaagtggcgg tcacagagat 6781 gacgccagct ctggtgcgtg cgtggtgggc cgggatgggt aggaagcacc cgactgcccg 6841 ccggcatgcc tacaacgtcc tccgggcggt gatgaacaca gcggtcgagg acaagctgat 6901 cgcagagaac ccgtgccgga tcgagcagaa ggcagccgat gagcgcgacg tagaggcgct 6961 gacgcctgag gagctggaca tcgtcgccgc tgagatcttc gtgcactacc ggatcgcggc 7021 atacatcctg gcgtggacga gcctccggtt cggagagctg atcgagcttc gccgcaagga 7081 catcgtggac gacggcatga cgatgaagct ccgggtgcgc cgtggcgctt cccgcgtggg 7141 gaacaagatc gtcgttggca acgccaagac cgtccggtcg aagcgtcctg tgacggttcc 7201 gcctcacgtc gcggagatga tccgagcgca catgaaggac cgtacgaaga tgaacaaggg 7261 ccccgaggca ttcctggtga ccacgacgca gggcaaccgg ctgtcgaagt ccgcgttcac 7321 caagtcgctg aagcgtggct acgccaagat cggtcggccg gaactccgca tccacgacct 7381 ccgcgctgtc ggcgctacgt tcgccgctca ggcaggtgcg acgaccaagg agctgatggc 7441 ccgcctcggt cacacgactc ctaggatggc gatgaagtac cagatggcgt ctgaggcccg 7501 cgacgaggct atcgctgagg cgatgtccaa gctggccaag acctcctgaa acgcaaaaag 7561 cccccctccc aaggacactg agtcctaaag aggggggttt cttgtcagta cgcgaagaac 7621 cacgcctggc cgcgagcgcc agcaccgccg ctctgtgcgg agacctgggc accagccccg 7681 ccgccgccag gagcattgcc gttcccgcca gctagccacc tgacgtcggg ggggggggaa 7741 agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca 7801 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt 7861 caacgggaaa cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat 7921 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat 7981 gggaagcccc atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat 8041 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc 8101 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa 8161 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg 8221 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat 8281 cgcgtatttc gcctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt 8341 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataat 8401 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt 8461 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac 8521 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag 8581 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat 8641 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc 8701 attacgctga cttgacggga cggcggcttt gttgaataaa tcgaactttt gctgagttga 8761 aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca 8821 aaatcaccaa ctggtccacc tacaacaaag ctctcaccaa ccgtggctcc ctcacgatat 8881 c //