| 出品公司: | ZYbscience |
|---|---|
| 别名: | pET23d, pet 23d |
| 质粒类型: | 大肠杆菌蛋白表达 |
| 表达水平: | 高 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 3663bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
| 载体标签: | N-T7, C-His |
| 载体抗性: | Ampicillin |
| 备注: |
Same as pET21abcd(+) but no lac; a,b,c,d vary by MCS.
|
| 产品目录号: | ZY69748-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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- 2025年07月14日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pET-23d(+)
- 库存：
20
- 供应商：
泽叶生物
pET23d载体基本信息
pET23d载体质粒图谱和多克隆位点信息
The maps for pET-23b(+), pET-23c(+) and pET-23d(+) are the same as pET-23a(+) (shown) with the following exceptions: pET-23b(+) is a 3665bp plasmid; subtract 1bp from each site beyond BamH I at 198. pET-23c(+) is a 3664bp plasmid; subtract 2bp from each site beyond BamH I at 198. pET-23d(+) is a 3663bp plasmid; the BamH I site is in the same reading frame as in pET-23c(+). An Nco I site is substituted for the Nde I site with a net 1bp deletion at position 238 of pET-23c(+). As a result, Nco I cuts pET-23d(+) at 234, and Nhe I cuts at 229. For the rest of the sites, subtract 3bp from each site beyond position 239 in pET-23a(+). Nde I does not cut pET-23d(+). Note also that Sty I is not unique in pET-23d(+).
pET23d载体简介
The pET-23a-d(+) vectors carry an N-terminal T7•Tag® sequence plus an optional C-terminal His•Tag® sequence. These vectors differ from pET-21a-d(+) by the “plain” T7 promoter instead of the T7lac promoter and by the absence of the lacI gene. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer.
pET23d载体序列
LOCUS pET-23d(+) 3663 bp DNA circular 11-SEP-2008
SOURCE
ORGANISM
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
promoter complement(300..316)
/vntifkey="30"
/label=T7\promoter
protein_bind complement(205..237)
/vntifkey="31"
/label=T7\tag
protein_bind complement(140..157)
/vntifkey="31"
/label=His\tag
terminator complement(26..72)
/vntifkey="43"
/label=T7\terminator
rep_origin complement(1447..1447)
/vntifkey="33"
/label=ColE1\origin
CDS complement(2208..3065)
/vntifkey="4"
/label=bla(amp)\sequence
rep_origin complement(3197..3652)
/vntifkey="33"
/label=f1\origin
BASE COUNT 918 a 935 c 926 g 884 t
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagccatggt
241 atatctcctt cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct
301 atagtgagtc gtattaattt cgcgggatcg agatctcggg cagcgttggg tcctggccac
361 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
421 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
481 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
541 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
601 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
661 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
721 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
781 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
841 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
901 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
961 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
1021 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
1081 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
1141 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
1201 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
1261 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
1321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
1381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
1441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
1501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
1561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
1621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
1681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
1741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
1801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
1861 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
1921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
1981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
2041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
2101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
2161 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
2221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
2281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
2341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
2401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
2461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
2521 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
2581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
2641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
2701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
2761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
2821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
2881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
2941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
3001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
3061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
3121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
3181 cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
3241 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
3301 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
3361 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
3421 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
3481 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
3541 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
3601 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
3661 cca
SOURCE
ORGANISM
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
promoter complement(300..316)
/vntifkey="30"
/label=T7\promoter
protein_bind complement(205..237)
/vntifkey="31"
/label=T7\tag
protein_bind complement(140..157)
/vntifkey="31"
/label=His\tag
terminator complement(26..72)
/vntifkey="43"
/label=T7\terminator
rep_origin complement(1447..1447)
/vntifkey="33"
/label=ColE1\origin
CDS complement(2208..3065)
/vntifkey="4"
/label=bla(amp)\sequence
rep_origin complement(3197..3652)
/vntifkey="33"
/label=f1\origin
BASE COUNT 918 a 935 c 926 g 884 t
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagccatggt
241 atatctcctt cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct
301 atagtgagtc gtattaattt cgcgggatcg agatctcggg cagcgttggg tcctggccac
361 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
421 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
481 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
541 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
601 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
661 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
721 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
781 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
841 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
901 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
961 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
1021 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
1081 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
1141 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
1201 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
1261 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
1321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
1381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
1441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
1501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
1561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
1621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
1681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
1741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
1801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
1861 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
1921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
1981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
2041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
2101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
2161 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
2221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
2281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
2341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
2401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
2461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
2521 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
2581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
2641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
2701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
2761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
2821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
2881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
2941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
3001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
3061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
3121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
3181 cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
3241 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
3301 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
3361 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
3421 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
3481 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
3541 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
3601 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
3661 cca
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文献和实验相关实验
加热灭活酶。通过凝胶提取试剂盒纯化消化的pET-32α（+）载体骨架和从pMD18-T载体切下的基因片段。用T4 DNA连接酶在23℃下用pET-32α（+）载体骨架连接基因片段1小时，载体末端和插入末端的比例为1：3（fmol）。将10μl连接反应与100μl感受态大肠杆菌 BL21（DE3）在冰上混合10分钟，并将混合物在42℃下孵育90秒。加入800μlLB并在37°C下孵育40分钟。涂在含有100μg/ ml氨苄青霉素的LB平板上。在37°C孵育过夜。通过菌落PCR筛选插入物的存在或制备
相关专题 1)噬菌体DE3溶源化的菌株如BL21(DE3)是最常用的表达菌株，噬菌体DE3是λ噬菌体的衍生株，构建好的表达载体 可以直接转入表达菌株中，诱导蛋白的表达方式与lac启动子一样都是IPTG诱导。 2)表达菌株BL21(DE3)上带有lacI基因，T7 RNA 聚合酶编码基因以及lacUV5启动子，lacUV5启动子可以启动T7 RNA 聚合酶的表达; 3)表达质粒 如pET-28质粒带有T7启动子和编码阻遏
中，现在Invitrogen已经克服了这个问题。在载体上多加四个碱基GTGG，就是这四个碱基使它可以与PCR产物定向连接。其作用机理见下图。 结合TOPO技术的有五个系列载体：pET100/D-TOPO®、pET101/D-TOPO®、pET102/D-TOPO®、pET151/D-TOPO®和pET200/D-TOPO®，这五个载体根据所带的标记、是否具有蛋白酶切位点而各有不同。详细信息参见下图。 结合TOPO技术的有五个系列载体：pET100/D
技术资料暂无技术资料 索取技术资料
pET-23d(+)载体
¥800
