出品公司: | EMD Biosciences |
---|---|
载体名称: | pETBlue-2 |
质粒类型: | 细菌表达 |
高拷贝/低拷贝: | 高拷贝 |
克隆方法: | -- |
启动子: | CMV |
载体大小: | 3600 bp |
5' 测序引物及序列: | T7 Fwd[TAATACGACTCACTATAGGG] |
3' 测序引物及序列: | -- |
载体标签: | His (Cterm), HSV (Cterm) |
载体抗性: | 氨苄青霉素 |
筛选标记: | -- |
克隆菌株: | -- |
宿主细胞(系): | -- |
备注: | 蓝/白标识;唯一MCS;His和HSV标记 |
产品目录号: | -- |
稳定性: | -- |
组成型/诱导型: | -- |
病毒/非病毒: | 非病毒 |
买家导航
pETBlue-2载体 pETBlue2质粒基本信息
pETBlue-2质粒图谱 pETBlue2载体图谱和pETBlue-2载体序列 pETBlue2质粒序列多克隆位点信息
pETBlue-2质粒 pETBlue2载体简介
pETBlue-2质粒序列 pETBlue2载体序列
LOCUS pETBlue-2 3653 bp ds-DNA circular SYN 23-OCT-2018 DEFINITION Vector for blue/white screening and for inducible bacterial expression of tagged proteins. ACCESSION . VERSION . KEYWORDS pETBlue-2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3653) AUTHORS MilliporeSigma (Novagen) TITLE Direct Submission JOURNAL Exported Wednesday, March 4, 2020 from SnapGene Viewer 3.1.4 http://www.snapgene.com COMMENT This vector encodes a start codon that is positioned for optimal expression. FEATURES Location/Qualifiers source 1..3653 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" promoter 1..19 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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)." CDS complement(54..491) /codon_start=1 /gene="lacZ (truncated)" /product="LacZ-alpha fragment of beta-galactosidase" /note="lacZ-alpha" /translation="MTMITTPNLINISGGGGGARDPLGLQERVLAGLHVYTAVRPQASN AYRWYRRRPAGAPVQNSDPRAPGISPWYISFLKLNKIILLANQDLSLAVVLQRRDWENP GVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL" RBS 265..270 /note="ribosome binding site" CDS 278..280 /codon_start=1 /product="start codon" /note="ATG" /translation="M" misc_feature 283..298 /note="blunt sites" /note="blunt cutter sites that allow ORF insertion in all three reading frames" misc_feature 377..392 /note="blunt sites" /note="blunt cutter sites that allow ORF insertion in all three reading frames" CDS 398..430 /codon_start=1 /product="HSV (herpes simplex virus) epitope tag" /note="HSV tag" /translation="QPELAPEDPED" CDS 437..454 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" promoter complement(541..569) /note="tet promoter" /note="E. coli promoter for tetracycline efflux protein gene" terminator 739..825 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 917..944 /note="rrnB T2 terminator" /note="transcription terminator T2 from the E. coli rrnB gene" terminator 1012..1059 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1096..1551 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(1666..2526) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 2619..3207 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 3606..3625 /bound_moiety="lac repressor encoded by lacI" /note="lac operator (symmetric)" /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). The symmetric lac operator was optimized for tight binding of lac repressor." ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gacttacaat 61 ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtacg ggcctcttcg 121 ctattacgcc agcttgcgaa cggtgggtgc gctgcaaggc gattaagttg ggtaacgcca 181 ggattctccc agtcacgacg ttgtaaaacg acggccagcg agagatcttg attggctagc 241 agaataattt tgtttaactt taagaaggag atataccatg gcgatatccc gggagctcgt 301 ggatccgaat tctgtacagg cgcgcctgca ggacgtcgac ggtaccatcg atacgcgttc 361 gaagcttgcg gccgcacagc tgtatacacg tgcaagccag ccagaactcg ctcctgaaga 421 cccagaggat ctcgagcacc accaccacca ccactaatgt taattaagtt gggcgttgta 481 atcatagtca taatcaatac tcctgactgc gttagcaatt taactgtgat aaactaccgc 541 attaaagcta ttcgatgata agctgtcaaa catgataatt cttgaagacg aaagggccta 601 ggctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc 661 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag 721 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt 781 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat 841 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc 901 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact 961 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctgagcaata actagcataa 1021 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc 1081 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 1141 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 1201 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 1261 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 1321 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 1381 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 1441 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 1501 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca 1561 cgatggcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 1621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 1681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 1741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 1801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 1861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 1921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 1981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 2041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 2101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 2161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 2221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 2281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 2341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 2401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 2461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 2521 actcatactc ttcctttttc aatcatgacc aaaatccctt aacgtgagtt ttcgttccac 2581 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 2641 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 2701 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 2761 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 2821 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 2881 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 2941 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 3001 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 3061 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 3121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 3181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 3241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 3301 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 3361 agcgagtcag tgagcgagga agccggcgat aatggcctgc ttctcgccga aacgtttggt 3421 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga 3481 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc 3541 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgac 3601 cggtgaattg tgagcgctca caattctcgt gacatcataa cgtcccgcga aat //
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pETBlue-2质粒 pETBlue2载体应用举例
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