质粒名称: | pCgVchcrRNA |
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出品公司: | Addgene |
目录编号: | 203815 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pJYS1Ptac |
完整质粒大小: | 6406 bp |
载体类型: | Bacterial Expression, 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 |
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插入基因名称: | CRISPR(BsaI) |
物种来源: | Vibrio cholera |
克隆方法: | Gibson Cloning |
文献引用方法 |
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材料和方法部分: | pCgVchcrRNA was a gift from Sheng Yang (Addgene plasmid # 203815 ; http://n2t.net/addgene:203815 ; RRID:Addgene_203815) |
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载体基本信息
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载体简介
载体序列
LOCUS pCgVchcrRNA 6406 bp ds-DNA circular SYN 07-AUG-2023 DEFINITION Expression of CRISPR RNA, shuttle vector for Corynebacterium glutamicum / E. coli. ACCESSION . VERSION . KEYWORDS pCgVchcrRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6406) TITLE Sheng Yang Plasmids 2023 REFERENCE 2 (bases 1 to 6406) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 7, 2023 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6406 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(100..894) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(211..230) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 821..840 /label=Neo-R /note="Neomycin resistance gene, reverse primer" rep_origin complement(3853..4591) /direction=LEFT /label=CloDF13 ori /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." terminator 4819..4846 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 4938..5023 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(5104..5727) /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 DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCKSGS" primer_bind 5635..5654 /label=Tet-R /note="Tetracycline resistance gene, reverse primer" protein_bind 6147..6165 /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O1 for the tetR and tetA genes" ORIGIN 1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 61 caaatatgta tccgctcatg aattaattca agcttcatat cagaagaact cgtcaagaag 121 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg 181 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg 241 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc 301 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg 361 catccgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc 421 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg 481 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc 541 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc 601 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc 661 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttggagttc 721 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag 781 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag 841 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa 901 cgatcctcat cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg 961 caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc 1021 tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag 1081 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca 1141 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg 1201 cttccggtga ccgcctcgat gatcgccggg tgggcgtggc ccaaggagga tcatccagcc 1261 attcggggtc gttcactggt tcccctttct gatttctggc atagaagaac ccccgtgaac 1321 tgtgtggttc cgggggttgc tgatttttgc gagacttctc gcgcaattcc ctagcttagg 1381 tgaaaacacc atgaaacact agggaaacac ccatgaaaca cccattaggg cagtagggcg 1441 gcttcttcgt ctagggcttg catttgggcg gtgatctggt ctttagcgtg tgaaagtgtg 1501 tcgtaggtgg cgtgctcaat gcactcgaac gtcacgtcat ttaccgggtc acggtgggca 1561 aagagaacta gtgggttaga cattgttttc ctcgttgtcg gtggtggtga gcttttctag 1621 ccgctcggta aacgcggcga tcatgaactc ttggaggttt tcaccgttct gcatgcctgc 1681 gcgcttcatg tcctcacgta gtgccaaagg aacgcgtgcg gtgaccacga cgggcttagc 1741 ctttgcctgc gcttctagtg cttcgatggt ggcttgtgcc tgcgcttgct gcgcctgtag 1801 tgcctgttga gcttcttgta gttgctgttc tagctgtgcc ttggttgcca tgctttaaga 1861 ctctagtagc tttcctgcga tatgtcatgc gcatgcgtag caaacattgt cctgcaactc 1921 attcattatg tgcagtgctc ctgttactag tcgtacatac tcatatttac ctagtctgca 1981 tgcagtgcat gcacatgcag tcatgtcgtg ctaatgtgta aaacatgtac atgcagattg 2041 ctgggggtgc agggggcgga gccaccctgt ccatgcgggg tgtggggctt gccccgccgg 2101 tacagacagt gagcaccggg gcacctagtc gcggataccc cccctaggta tcggacacgt 2161 aaccctccca tgtcgatgca aatctttaac attgagtacg ggtaagctgg cacgcatagc 2221 caagctaggc ggccaccaaa caccactaaa aattaatagt ccctagacaa gacaaacccc 2281 cgtgcgagct accaactcat atgcacgggg gccacataac ccgaaggggt ttcaattgac 2341 aaccatagca ctagctaaga caacgggcac aacacccgca caaactcgca ctgcgcaacc 2401 ccgcacaaca tcgggtctag gtaacactga aatagaagtg aacacctcta aggaaccgca 2461 ggtcaatgag ggttctaagg tcactcgcgc tagggcgtgg cgtaggcaaa acgtcatgta 2521 caagatcacc aatagtaagg ctctggcggg gtgccatagg tggcgcaggg acgaagctgt 2581 tgcggtgtcc tggtcgtcta acggtgcttc gcagtttgag ggtctgcaaa actctcactc 2641 tcgctggggg tcatctctgg ctgaattgga agtcatgggc gaacgccgca ttgagctggc 2701 tattgctact aagaatcact tggcggcggg tggcgcgctc atgatgtttg tgggcactgt 2761 tcgacacaac cgctcacagt catttgcgca ggttgaagcg ggtattaaga ctgcgtactc 2821 ttcgatggtg aaaacatctc agtggaagaa agaacgtgca cggtacgggg tggagcacac 2881 ctatagtgac tatgaggtca cagactcttg ggcgaacggt tggcacttgc accgcaacat 2941 gctgttgttc ttggatcgtc cactgtctga cgatgaactc aaggcgtttg aggattccat 3001 gttttcccgc tggtctgctg gtgtggttaa ggccggtatg gacgcgccac tgcgtgagca 3061 cggggtcaaa cttgatcagg tgtctacctg gggtggagac gctgcgaaaa tggcaaccta 3121 cctcgctaag ggcatgtctc aggaactgac tggctccgct actaaaaccg cgtctaaggg 3181 gtcgtacacg ccgtttcaga tgttggatat gttggccgat caaagcgacg ccggcgagga 3241 tatggacgct gttttggtgg ctcggtggcg tgagtatgag gttggttcta aaaacctgcg 3301 ttcgtcctgg tcacgtgggg ctaagcgtgc tttgggcatt gattacatag acgctgatgt 3361 acgtcgtgaa atggaagaag aactgtacaa gctcgccggt ctggaagcac cggaacgggt 3421 cgaatcaacc cgcgttgctg ttgctttggt gaagcccgat gattggaaac tgattcagtc 3481 tgatttcgcg gttaggcagt acgttctcga ttgcgtggat aaggctaagg acgtggccgc 3541 tgcgcaacgt gtcgctaatg aggtgctggc aagtctgggt gtggattcca ccccgtgcat 3601 gatcgttatg gatgatgtgg acttggacgc ggttctgcct actcatgggg acgctactaa 3661 gcgtgatctg aatgcggcgg tgttcgcggg taatgagcag actattcttc gcacccacta 3721 aaagcggcat aaaccccgtt cgatattttg tgcgatgaat ttatggtcaa tgtcgcgggg 3781 gcaaactatg atgggtcttg ttgtttagct agctcactcg gtcgctacgc tccgggcgtg 3841 agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt ggataagcag 3901 gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc cataggctcc 3961 gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg ggagccgtga 4021 ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag atccccaccg 4081 tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt taccggatac 4141 ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca cacactggta 4201 tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac tccccgttca 4261 gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg aaaagcacgg 4321 taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt tgtttagcta 4381 aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag gacagatttg 4441 gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac tgacttaacc 4501 ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag 4561 aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt 4621 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt 4681 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag 4741 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc 4801 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc 4861 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc 4921 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata 4981 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttaaattct agactgatga 5041 atcccctaat gatttttatc aaaatcatta aggttaccat cacggaaaaa ggttatgctg 5101 cttttaagac ccacttttac atttaagttg tttttctaat ccgcatatga tcaattcaag 5161 gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa 5221 taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc 5281 ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc 5341 attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata 5401 ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa 5461 agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa 5521 agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc 5581 ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag 5641 tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat 5701 cactttactt ttatctaatc tagacatgtt gtgtctcctc taaagattgt aggaaatgca 5761 atgtgtcaag cagttaggtc aatcagagca tcggtcactc aagaattctt gagctgacag 5821 aacccaaggg cgagtatggc gaatttttcc aactcacgca ccaaacctac tcacattgga 5881 taattcgtcg agtcaactcc gtgtcaacct gtcctcaact ttagcgggga aacaaagatt 5941 tcgcagcaga tgcaaaatca tacagacaca ttcaaaaatg gacattgacc ccaaacgggg 6001 gcatttccaa cacttgcgct aaatagtttt tgcagggaac gaacctgaaa tcgttcgcac 6061 tacgatctgc tcccccgaat ggggttaatt atgaactaca ttacaccccc gtgaatcgaa 6121 ttccaaaaac taaaaaaaat attgacactc tatcattgat agagtataat taacgggatc 6181 cactcttcat tgataagtcg ccaagcttgg gatccccagc ttgttgatac actaatgctt 6241 ttatataggg aaaaggtggt gaactactcc atgggtgaac tgccgagtag gtagctgata 6301 acgagacctc tggtctcgtg aactgccgag taggtagctg ataacggatc cgaattcgag 6361 ctcggcgcgc ctgcaggtcg acaagcttgc gctcgagcgg gggccc //
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