质粒名称: | pSY14-ptb |
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出品公司: | Addgene |
目录编号: | 90490 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pSY8 |
完整质粒大小: | 4751bp |
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: | 未知 |
插入基因信息 |
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插入基因名称: | sceC |
克隆方法: | 限制性内切酶 |
5’克隆位点: | unknown (unknown if destroyed) |
3’克隆位点: | unknown (unknown if destroyed) |
文献引用方法 |
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材料和方法部分: | pSY14-ptb was a gift from Sheng Yang (Addgene plasmid # 90490 ; http://n2t.net/addgene:90490 ; RRID:Addgene_90490) |
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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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 pSY14-ptb 4751 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pSY8 derivative with sceC gene under the control of ptb promoter used for I-SceI-mediated scarless gene modification via allelic exchange in Clostridium. ACCESSION . VERSION . KEYWORDS pSY14-ptb SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4751) 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 4751) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4751 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(356..373) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(527..1115) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(607..626) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1286..2146) /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 1909..1928 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2147..2251) /gene="bla" /label=AmpR promoter primer_bind 2319..2337 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2375..2397) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2497..2516 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cccttgtcac taagaaaata aatgcagggt aaaatttata tccttcttgt tttatgtttc 61 ggtataaaac actaatatca atttctgtgg ttatactaaa agtcgtttgt tggttcaaat 121 aatgattaaa tatctctttt ctcttccaat tgtctaaatc aattttatta aagttcattt 181 gatatgcctc ctaaattttt atctaaagtg aatttaggag gcttacttgt ctgctttctt 241 cattagaatc aatccttttt taaaagtcaa tattactgta acataaacag ctgcattaat 301 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 361 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 421 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 481 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 541 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 601 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 661 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 721 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 781 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 841 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 901 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 961 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 1021 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 1081 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 1141 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 1201 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 1261 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 1321 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 1381 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 1441 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 1501 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 1561 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 1621 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 1681 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 1741 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 1801 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 1861 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 1921 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 1981 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 2041 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 2101 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 2161 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 2221 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 2281 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct 2341 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga 2401 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag 2461 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga 2521 gagtgcacca tatgctcgag ttatttaaga aaagtttcac tactaattgt atttggaagt 2581 ttatacatca tttgtggaat aagatatggt ttaattaaat tgtaaaatat aagatatgac 2641 atactatcaa tataaattat aggtttattt ttatttattt ttacataaca attaagttga 2701 aatttatttc taagtccttt tactaaatat tcaacttctt caaaagtaaa agattgtgta 2761 tttaaaacta tacttttatt agtagaattt ttattataat cccatttacc tccatcatcc 2821 ataaaccaat atgcaagtga cattggagta agataatttt ctactaaatt attaggaatt 2881 gttttcttat tattaacaat aaaaagatta gcaagtttat taaatgcttg atgtttaaaa 2941 gtttgagcac cccatgtaat tacaagattt ccaagatgat taactctttg ttttttatgt 3001 ggaggtgaaa gtacccattg atcataaaga agacaaacat gatccatata tgctttattt 3061 ttccattcaa attgcataca atatgtttta ccttcatctc tacttcttat ataagcatct 3121 ccaagaataa gaccaattcc tgcttcaaat tgttctatat taagttctat aagttgtgat 3181 ttatattctt taagaagttt actattagga ccaagattca ttacttgatt cttttgatgc 3241 atatgtcgac actccctttt actatttaat tatctatgtt aaatgattaa cgtgctcatt 3301 tatattttaa caaacttttc acatgaagtt aagtttttta gaaaattatt tatattttat 3361 agaattcccg gggatccgtc gacctgcagc caagcttaat cgcatttcat agattgacct 3421 cccaataact acgtggtgtt attgggaggt caatctattt catttgcctc ttgctcaaag 3481 ttcccaaatt cgagtaagag gtatttttgt ttttggtcgt cgcctctcat tagtagttca 3541 gggtttaaca ttaatactcc agtttttctt tttataatat ttccttcttc taagatttta 3601 agtgttgtta ttactgtttg tagacttgtt cctgtagctt ttgctatttc tcttgttgta 3661 gctatcattg tattgttact taagtggaca ttatctagga tatagttaac gattttaagt 3721 ttttttccgc caatcatatc taacatactt attaattgca ctatatatgc ctttacgaag 3781 ttaccagacg tttgtttacg gtataacttg tctacctcta tgacttctcc actttcttcg 3841 tctatgagcc tctgagagcc tttatagact gttccatatc tttctttcat ctttttctca 3901 ctccttattt taaactattc taactatatc ataactgttc taaaaaaaaa agaacatttg 3961 ttaaaagaaa ttagaacaaa atgagtgaaa aattagaaca aacaaattcc ttataaacct 4021 tatcatctca acctatatta agattttacc tagttgaatc ttcttttcta tataaagcgt 4081 cggagcatat cagggggtta tctaacgtaa atgctaccct tcggctcgct ttcgctcggc 4141 attgacgtca gatactgcac cccctgaacc cccatgctcc aacagcaaaa aggaaacttt 4201 ttgctgcttt tccgacgctt attcgcttcg ctcatattta tatagaaaag aagtgaatgc 4261 gcaaaagaca taatcgattt ataaaagcca gtcattaggc ctatctgaca attcctgaat 4321 agagttcata aacaatcctg catgataacc atcacaaaca gaatgatgta cctgtaaaga 4381 tagcggtaaa tatattgaat tacctttatt aatgaatttt cctgctgtaa taatgggtag 4441 aaggtaatta ctattattat tgatatttaa gttaaaccca gtaaatgaag tccatggaat 4501 aatagaaaga gaaaaagcat tttcaggtat aggtgttttg ggaaacaatt tccccgaacc 4561 attatatttc tctacatcag aaaggtataa atcataaaac tctttgaagt cattctttac 4621 aggagtccaa ataccagaga atgttttaga tacaccatca aaaattgtat aaagtggctc 4681 taacttatcc caataaccta actctccgtc gctattgtaa ccagttctaa aagctgtatt 4741 tgagtttatc a //
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