pET-29b(+)
pET-29b(+)
编号 | 名称 |
北京华越洋VECT-650 | pET-29b(+) |
pET29b载体基本信息
别名: | pET29b, pet 29b |
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: | 5370 bp |
5' 测序引物: | T7 |
5' 测序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
载体标签: | N-S, N-Thrombin, C-His |
载体抗性: | Kanamycin |
备注: | N term 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 |
pET29b载体质粒图谱和多克隆位点信息
Feature Name | Start | End |
T7_terminator | 129 | 1 |
T7_Terminal_primer | 69 | 87 |
lacI | 217 | 1856 |
T7_transl_en_RBS | 320 | 304 |
lacO | 365 | 338 |
T7_promoter | 383 | 365 |
tet (300 - 563) | 419 | 682 |
pBRrevBam_primer | 490 | 471 |
tet (576 - 851) | 1915 | 2190 |
ROP | 2665 | 2856 |
pGEX_3_primer | 2872 | 2850 |
pBR322_origin | 3890 | 3271 |
KanR2 | 3996 | 4811 |
ORF | Start | End |
ORF frame 2 | 296 | 1024 |
ORF frame 3 | 897 | 1856 |
ORF frame 3 | 3996 | 4811 |
Enzyme Name | Cut |
XhoI | 158 |
NotI | 166 |
EagI | 166 |
HindIII | 173 |
SalI | 179 |
SacI | 190 |
EcoRI | 192 |
BamHI | 198 |
EcoRV | 206 |
NcoI | 211 |
KpnI | 237 |
BglII | 240 |
BstBI | 267 |
NdeI | 294 |
XbaI | 332 |
BclI | 1138 |
ApaI | 1335 |
HpaI | 1630 |
FspI | 2206 |
NruI | 4084 |
SmaI | 4301 |
XmaI | 4299 |
pET29b载体简介
The pET-29a-c(+) vectors carry an N-terminal S•Tag™/thrombin 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 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.
pET29b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 tccgatatcg ccatggaacc gcgtggcacc agggtaccca
241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcatatgta
301 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
361 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc
421 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
481 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
541 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
601 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
661 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
721 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
781 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
841 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
901 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
961 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
1021 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
1081 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
1141 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
1201 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
1261 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
1321 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
1381 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
1441 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
1501 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
1561 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
1621 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
1681 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
1741 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
1801 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
1861 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg
1921 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
1981 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
2041 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
2101 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
2161 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc
2221 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
2281 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
2341 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
2401 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
2461 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
2521 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
2581 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
2641 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
2701 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
2761 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
2821 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
2881 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
2941 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
3001 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga
3061 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
3121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
3181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
3721 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac
3961 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
4021 cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
4081 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
4141 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
4201 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
4261 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
4321 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
4381 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
4441 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
4501 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
4561 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
4621 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
4681 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
4741 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
4801 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa
4861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
4921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
4981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
5041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
5101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
5161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
5221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
5281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
5341 gcgccgctac agggcgcgtc ccattcgcca
//
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