pET-28c(+)
pET-28c(+)
编号 | 名称 |
北京华越洋VECT-360 | pET-28c(+) |
pET28c载体基本信息
别名: | pET28c, pet 28c |
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: | 5367 bp |
5' 测序引物及序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
3' 测序引物及序列: | T7t: 5'-GCTAGTTATTGCTCAGCGG-3' |
载体标签: | N-His, N-Thrombin, N-T7, C-His |
载体抗性: | Kanamycin |
备注: | Nterm thrombin cleavage site; a,b,c vary by MCS; |
稳定性: | 瞬时表达 Transient |
组成型: | 组成型 Constitutive |
病毒/非病毒: | 非病毒 |
pET28c载体质粒图谱和多克隆位点信息
pET28c载体简介
pET-28a-c(+)载体带有一个N端的His/Thrombin/T7蛋白标签,同时含有一个可以选择的C端His标签。pET28c载体的单一的多克隆位点见上面的环状质粒图谱。
注意:载体序列是以pBR322质粒的编码规矩进行编码的,所以T7蛋白表达区在质粒图谱上面是反向的。 T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的,所以在T7噬菌体聚合酶的作用下,包含有蛋白编码序列的病毒粒子能够产生,并启动蛋白表达,同时蛋白表达将被T7终止子序列(Cat. No. 69337-3)的作用下终止蛋白翻译。
The pET-28a-c(+) vectors carry an N-terminal His•Tag®/ thrombin/ T7•Tag® configuration plus an optional C-terminal His•Tag sequence. 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 polymeraseis 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 (Cat. No. 69337-3).
pET28c载体序列
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 tagccatatg
241 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt
301 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
361 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
421 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
481 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
541 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
601 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
661 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
721 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
781 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
841 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
901 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
961 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
1021 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
1081 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
1141 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
1201 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
1261 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
1321 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
1381 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
1441 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
1501 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
1561 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
1621 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
1681 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
1741 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
1801 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
1861 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
1921 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
1981 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
2041 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
2101 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
2161 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
2221 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
2281 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
2341 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
2401 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
2461 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
2521 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
2581 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
2641 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
2701 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
2761 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
2821 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
2881 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
2941 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
3001 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
3061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
3121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
3181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
3241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
3301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
3361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
3421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
3481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
3541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
3601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
3661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
3721 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
3781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
3841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
3901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaaca ataaaactgt
3961 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
4021 gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
4081 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
4141 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
4201 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
4261 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
4321 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
4381 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
4441 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
4501 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
4561 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
4621 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
4681 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
4741 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
4801 ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa
4861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt
4921 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
4981 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
5041 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
5101 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
5161 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
5221 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
5281 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
5341 ccgctacagg gcgcgtccca ttcgcca
//
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