质粒名称: | pBAD-I-SceI |
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出品公司: | addgene |
目录编号: | 60960 |
存储实验室: | Nikolaus Osterrieder |
载体骨架基本信息 |
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载体名称: | pBAD24 |
完整质粒大小: | 5238 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH10B |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | I-SceI restriction enzyme |
文献引用方法 |
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材料和方法部分: | pBAD-I-SceI was a gift from Nikolaus Osterrieder (Addgene plasmid # 60960 ; http://n2t.net/addgene:60960 ; RRID:Addgene_60960) |
参考文献部分: | Two-step red-mediated recombination for versatile high-efficiency markerless DNA manipulation in Escherichia coli. Tischer BK, von Einem J, Kaufer B, Osterrieder N. Biotechniques. 2006 Feb;40(2):191-7. 10.2144/000112096 PubMed 16526409 |
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载体序列
LOCUS pBAD-I-SceI 5238 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid that allows an arabinose-inducible expression of the I-SceI homing endonuclease - for the en passant (two-step Red-mediated) mutagenesis . ACCESSION . VERSION . KEYWORDS pBAD-I-SceI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5238) AUTHORS Tischer BK, von Einem J, Kaufer B, Osterrieder N TITLE Two-step red-mediated recombination for versatile high-efficiency markerless DNA manipulation in Escherichia coli. JOURNAL Biotechniques. 2006 Feb;40(2):191-7. PUBMED 16526409 REFERENCE 2 (bases 1 to 5238) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..5238 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..63) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 216..302 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 394..421 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 440..531 /gene="bla" /label=AmpR promoter CDS 532..1392 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(750..769) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1434..1889 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1521..1540) /label=F1ori-R /note="F1 origin, reverse primer" rep_origin 2000..2588 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2489..2508 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2742..2759 /label=L4440 /note="L4440 vector, forward primer" primer_bind 2925..2947 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3086..3105 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(3285..4163) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 4190..4474 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 4397..4416 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" ORIGIN 1 ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa 61 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt 121 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg 181 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga 241 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa 301 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac 361 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt 421 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc 481 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 541 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 601 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 661 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 721 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac 781 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 841 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 901 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 961 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 1021 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 1081 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 1141 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 1201 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 1261 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 1321 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 1381 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc 1441 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 1501 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 1561 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 1621 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc 1681 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 1741 gttccaaact tgaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat 1801 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 1861 ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata 1921 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 1981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 2041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 2101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 2161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 2221 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 2281 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 2341 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 2401 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 2461 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 2521 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 2581 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 2641 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 2701 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 2761 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 2821 gcatagggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc 2881 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg 2941 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag caaggagatg gcgcccaaca 3001 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga 3061 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac 3121 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga 3181 cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc 3241 ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg 3301 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg 3361 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg 3421 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc 3481 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac 3541 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga 3601 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg 3661 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc 3721 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc 3781 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta 3841 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga 3901 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc 3961 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga 4021 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg 4081 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg 4141 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat 4201 tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc 4261 aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca 4321 aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac 4381 ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga 4441 cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc tagcaggagg 4501 gtacctatat gcatatgaaa aacatcaaaa aaaaccaggt aatgaacctg ggtccgaact 4561 ctaaactgct gaaagaatac aaatcccagc tgatcgaact gaacatcgaa cagttcgaag 4621 caggtatcgg tctgatcctg ggtgatgctt acatccgttc tcgtgatgaa ggtaaaacct 4681 actgtatgca gttcgagtgg aaaaacaaag catacatgga ccacgtatgt ctgctgtacg 4741 atcagtgggt actgtccccg ccgcacaaaa aagaacgtgt taaccacctg ggtaacctgg 4801 taatcacctg gggcgcccag actttcaaac accaagcttt caacaaactg gctaacctgt 4861 tcatcgttaa caacaaaaaa accatcccga acaacctggt tgaaaactac ctgaccccga 4921 tgtctctggc atactggttc atggatgatg gtggtaaatg ggattacaac aaaaactcta 4981 ccaacaaatc gatcgtactg aacacccagt ctttcacttt cgaagaagta gaatacctgg 5041 ttaagggtct gcgtaacaaa ttccaactga actgttacgt aaaaatcaac aaaaacaaac 5101 cgatcatcta catcgattct atgtcttacc tgatcttcta caacctgatc aaaccgtacc 5161 tgatcccgca gatgatgtac aaactgccga acactatctc ctccgaaact ttcctgaaat 5221 aagggcccga cgtctgca //
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