| 出品公司: | Novagen |
|---|---|
| 载体名称: | pET-35b（+） |
| 质粒类型: | 大肠杆菌表达 |
| 启动子: | T7 |
| 载体大小: | 5927bp |
| 5' 测序引物及序列: | T7:TAATACGACTCACTATAGGG |
| 3' 测序引物及序列: | T7-ter:TGCTAGTTATTGCTCAGCGG |
| 载体标签: | N-CBDclos,C-6×His |
| 载体抗性: | 卡那霉素 |
| 克隆菌株: | DH5α 等 |
| 表达宿主: | BL21(DE3) |
| 培养条件: | 37℃，有氧，LB |
| 诱导方式: | IPTG |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3) |
| pET表达手册： |
pET表达手册英文版 pET表达手册中文版 |
| pET载体汇总: |
pET-3a pET-11a pET-12a pET-14b pET-15b pET-16b pET-17b pET-19b pET-20b pET-21a pET-22b pET-23a pET-24a pET-25b pET-26b pET-27b pET-28a pET-29a pET-30a pET-31b pET-32a pET-33b pET-34b pET-35b pET-37b pET-39b pET-40b pET-41a pET-42a pET-43.1a pET-44a pET-45b pET-47b pET-48b pET-49b pET-50b pET-51b pET-52b pETDuet-1 pACYCDuet-1 pCDFDuet-1 pRSFDuet-1 pColaDuet-1 |
买家导航
pET-35b(+)载体 pET35b质粒基本信息
pET-35b(+)质粒图谱 pET35b载体图谱和pET-35b(+)载体序列 pET35b质粒序列多克隆位点信息
pET-35b(+)质粒 pET35b载体简介
pET-35b(+)(Cat. No. 70103-3) 号载体是为表达cbdclos融合蛋白而设计的有多种克隆地点和策略可供选择。pCR扩增的快速定向克隆pET-35xa/lic载体试剂盒(Cat. No. 70115-3)，包含线性化pET-35B（+）已准备好，可使用适当制备的插件进行退火。当用xa/lic克隆时产生的cbdclos•tag™融合蛋白可以在编码的载体上精确地切割使用因子xa连接。独特的地点显示在圆形地图上。请注意，序列是按PBR322约定编号的，因此T7表达式区域在圆映射上反转。这个 由t7 rna聚合酶转录的编码链的克隆/表达区域如下所示。f1基因的起源是定向的，因此与辅助噬菌体的感染将产生含有单个与编码链相对应的链状DNA。因此，单链测序应该使用T7终端底漆(Cat. No. 69337-3)。
pET-35b(+)质粒序列 pET35b载体序列
LOCUS pET-35b 5927 bp ds-DNA circular SYN 07-8-2015 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Untitled SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5927) AUTHORS . TITLE Direct Submission JOURNAL Exported 2015-8-7 from MLCC http://www.miaolingbio.com FEATURES Location/Qualifiers source 1..5927 /organism="synthetic DNA construct" /mol_type="other DNA" source 860..882 /organism="Enterobacteria phage T7" /mol_type="genomic DNA" /db_xref="taxon:10760" terminator 26..73 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(150..173) /codon_start=1 /product="8xHis affinity tag" /note="8xHis" /translation="HHHHHHHH" CDS complement(249..260) /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" CDS complement(291..335) /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" CDS complement(351..368) /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" RBS 860..882 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 897..921 /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)." promoter complement(922..940) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" promoter 1253..1330 /gene="lacI" /note="lacI promoter" CDS 1331..2413 /codon_start=1 /gene="lacI" /product="lac repressor" /note="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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" CDS 3222..3413 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" misc_feature 3515..3657 /note="bom" /note="basis of mobility region from pBR322" rep_origin complement(3843..4431) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS 4553..5368 /codon_start=1 /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin complement(5461..5916) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt 181 gcggccgcaa gcttgtcgac ggagctcgaa ttcggatccg atatcgccat ggccagagga 241 gagttagagc gaccctcaat accggagcca ccaccggtac ccagatctgg gctgtccatg 301 tgctggcgtt cgaatttagc agcagcggtt tctttcatac caattgcagt actaccgcgt 361 ggcaccagac ccgcggaacc tggtgctgta ccaagaactt tagctccacc tatatatcct 421 gtaacttttg gatttacaac tggtgttgaa ctacttgcat caaatgaata gtcatttgtt 481 tgagtgtagt ttgaccagtc tgattttgtt attcttcctt gaatagttat aaattgtcct 541 tttttaagag tagctgctcc gcttgcaaat ccaaattcaa catatgtatc ataggttgat 601 gttgggcttg ctgtttcttt aacgaagttt gctgtcactt tgctagtgtt atcaacatag 661 ctatttccta ataatgcacc agcatggtca caccagaaag tttgtccttg tgtaccatca 721 cttgtgtaat aatatctaac ttttacgtca tttaaattta aatcactgtc agatgtgtta 781 gtaattttga ttattggtgt aattgagttt gtttgtgctg atttgttaga gttgtaaaat 841 tcaactgaca ttatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa 901 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg 961 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt 1021 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc 1081 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc 1141 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta 1201 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat 1261 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag 1321 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta 1381 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa 1441 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc 1501 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc 1561 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc 1621 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca 1681 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga 1741 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa 1801 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt 1861 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg 1921 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga 1981 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat 2041 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat 2101 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga 2161 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg 2221 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca 2281 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc 2341 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 2401 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat 2461 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca 2521 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc 2581 cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg 2641 gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc 2701 ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg 2761 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg 2821 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt 2881 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa 2941 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc 3001 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt 3061 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc 3121 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca 3181 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa 3241 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca 3301 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg 3361 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 3421 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 3481 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg 3541 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 3601 ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc 3661 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 3721 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 3781 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 3841 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 3901 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 3961 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 4021 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 4081 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 4141 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 4201 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 4261 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 4321 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 4381 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 4441 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 4501 gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca 4561 tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg atgctgattt 4621 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt 4681 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa 4741 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac 4801 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg 4861 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc 4921 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag 4981 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc 5041 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca 5101 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa 5161 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc 5221 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt 5281 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt 5341 tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac atatttgaat 5401 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 5461 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 5521 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 5581 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 5641 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct 5701 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc 5761 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag 5821 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca 5881 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgcca //
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pET-35b(+)质粒 pET35b载体应用举例
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