质粒名称: | pXY1-spc |
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出品公司: | Addgene |
目录编号: | 90488 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pXY1 |
完整质粒大小: | 5286 bp |
载体类型: | 细菌表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | 无 |
文献引用方法 |
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材料和方法部分: | pXY1-spc was a gift from Sheng Yang (Addgene plasmid # 90488 ; http://n2t.net/addgene:90488 ; RRID:Addgene_90488) |
参考文献部分: | I-SceI-mediated scarless gene modification via allelic exchange in Clostridium. Zhang N, Shao L, Jiang Y, Gu Y, Li Q, Liu J, Jiang W, Yang S. J Microbiol Methods. 2015 Jan;108:49-60. doi: 10.1016/j.mimet.2014.11.004. Epub 2014 Nov 15. 10.1016/j.mimet.2014.11.004 PubMed 25451462 |
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LOCUS pXY1-spc 5286 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid with ptb promoter for gene expression in Clostridium acetobutylicum ATCC 824 and Clostridium beijerinckii NCIMB8052. ACCESSION . VERSION . KEYWORDS pXY1-spc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5286) AUTHORS Zhang N, Shao L, Jiang Y, Gu Y, Li Q, Liu J, Jiang W, Yang S TITLE I-SceI-mediated scarless gene modification via allelic exchange in Clostridium. JOURNAL J Microbiol Methods. 2015 Jan;108:49-60. doi: 10.1016/j.mimet.2014.11.004. Epub 2014 Nov 15. PUBMED 25451462 REFERENCE 2 (bases 1 to 5286) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..5286 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(126..714) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(206..225) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(885..1745) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1508..1527 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1746..1850) /gene="bla" /label=AmpR promoter primer_bind 1918..1936 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(1974..1996) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2096..2115 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5010..5026) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5010..5026) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5023..5045) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5034..5050 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5058..5088) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5103..5124 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5241..5258) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 61 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 1021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 1081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 1141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 1201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 1261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 1321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 1381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 1441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 1501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 1561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 1621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 1681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 1741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 1801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 1861 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 1921 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 1981 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 2041 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 2101 tcagagcaga ttgtactgag agtgcaccat atgtcgacac tcccttttac tatttaatta 2161 tctatgttaa atgattaacg tgctcattta tattttaaca aacttttcac atgaagttaa 2221 gttttttaga aaattattta tattttatac tgcagcacat taagtatata ctattattaa 2281 actattctat atacttaatt tattttaata gaaaaacata atatcataat aacttcaaaa 2341 ttaaacttta tttatgattt catacttgac tttgatttta gaaaggatat actttttagc 2401 agatttggaa acggctttgg acgtagtttg cccatagatg aacaaacaaa ctacatccaa 2461 aaattatact tttcccttca ttggtatccg tatttttaca tcttaatagc gtatgtatta 2521 caacacacct aaacaacgac cttacggtct gctactgcat atcctagctt gattgtttag 2581 ttgcctcaac tatgcttaac cctaccccga actctttttt tattgtgggt tttcgtcgtg 2641 aagtcccacc gacacataat cataacataa gatgtattat gaaaatgcga gtgactatcc 2701 ttttgtatcg gctcactaca ccacagatat attttttagt gcatactgtg tcggcactct 2761 caatattaaa ttaataaata aattattttt tctttttact cttctttaca tgagcttttt 2821 taaagctcct tgcataatat ttaatgcatg tacgttcttt tttctgttct tcctctgtaa 2881 aacatctcat ttttatggca caccatccat atcggttcat ctttgaacaa ttaatacatt 2941 ggactttccc tttatgtaaa catcttgact cattgtattt actgcaatag gttgctactg 3001 ttttatcatg aatcatcaac gaatgtattt tgcatatttc agtatcttta ataaactctt 3061 tgcaattttt acaagttttc atatacgccc ttcttttcat aaattaattt atgaattcta 3121 tgtattccaa ggagcttttt aaagctcccc ctttcgtact acttagctac aagcactata 3181 aaagtcataa tgtttactgc taaagtcaat aatgatattg ctaatacctt gttatttgat 3241 aagatactgc tttcctctgt cactttgctc accccctttc attttcataa attaattatg 3301 aaaaaataaa tatacttcta atttttatca aataaaaaag cctttgcgta ctgcttccaa 3361 tacacaaagg ctataaactt ctaaatctta cttattgcaa tttacattta tatctgttaa 3421 gataatctca taaattgaat atatatagtt aatgtttctc ttgtatgtcg gtacatttga 3481 aatattgcta tagatagagt tctctaacgg cttgatgtgt tggtagcaca tttaagtttt 3541 ggcttatata ctaagttggt agcttaatat ataagagctg aggacttatt tttttattaa 3601 atttttcaac ttgtctatat tttaacccgt aattgaatac ataacaagta tttttttttg 3661 tattcaatta aacattcata aatgagtata attaatcata ctaaattcta taattttctt 3721 ttctgtaaat tatcgattaa tttttttaat ctgttattta aatagtttat agttaaattt 3781 acattttcat tagtccattc aatattctct ccaagataac tacgaactgc taacaaaatt 3841 ctctccctat gttctaatgg agaagattca gccactgcat ttcccgcaat atcttttggt 3901 atgattttac ccgtgtccat agttaaaatc atacggcata aagttaatat agagttggtt 3961 tcatcatcct gataattatc tattaattcc tctgacgaat ccataatggc tcttctcaca 4021 tcagaaaatg gaatatcagg tagtaattcc tctaagtcat aatttccgta tattctttta 4081 ttttttcgtt ttgcttggta aagcattatg gttaaatctg aatttaattc cttctgagga 4141 atgtatcctt gttcataaag ctcttgtaac cattctccat aaataaattc ttgtttggga 4201 ggatgattcc acggtaccat ttcttgctga ataataattg ttaattcaat atatcgtaag 4261 ttgcttttat ctcctatttt ttttgaaata ggtctaattt tttgtataag tatttcttta 4321 ctttgatctg tcaatggttc agatacgacg actaaaaagt caagatcact atttggtttt 4381 agtccactct caactcctga tccaaacatg taagtaccaa taaggttatt ttttaaatgt 4441 ttccgaagta tttttttcac tttattaatt tgttcgtatg tattcaaata tatcctcctc 4501 actattttga ttagtaccta ttttatatcc atagttgtta attaaataaa cttaatttag 4561 tttatttata gatttcattg gcttctaaat tttttatcta gataataatt attttagtta 4621 attttattct agattatata tgatatgatc tttcatttcc ataaaactaa agtaagtgta 4681 aacctattca ttgtggctct gataaatatg aacatgatga gtgatcgtta aatttatact 4741 gcaatctgat gcgattattg aataaaagat atgagagatt tatctagttt ctttttttac 4801 aagaaaaaag aaagttctta aaggttttat acttttggtc gtagagcaca cggtttaacg 4861 acttaattac gaagtaaata agtctagtgt gttagacttt aatgtttttt taaggcatta 4921 gtgcatttaa gcgtcagagc atggctttat gccgagaaaa ctattggttg gaatggcgtg 4981 tgtgttagcc aaagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 5041 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 5101 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 5161 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 5221 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 5281 cgagcg //
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