| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 别名: | pET43.1 EK/LIC, pET 43.1 EK/LIC |
| 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: | 7295 bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
| 3' 测序引物: | ColiDOWN |
| 3' 测序引物序列: | ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3' |
| 载体标签: | C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK |
| 载体抗性: | Ampicillin |
| 备注: |
For directional cloning of PCR-amplified DNA; ligation independent cloning; enterokinase cleavage site
|
| 产品目录号: | 71072-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3)、C43(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-43.1EK/LIC载体基本信息
pET-43.1EK/LIC载体图谱质粒图谱和多克隆位点信息
pET-43.1EK/LIC载体简介
pET-43.1a EK/LIC载体是用来快速定向PCR高通量质粒构建的载体。本载体带有N-Nus标签和His标签，S标签。独特的酶切位点在载体图谱中被标注出来。需要注意的是，这些载体序列是依据载体pBR322的方向来进行编号的，所以T7标签表达区域在载体图谱上是反向的。
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。
pET-43.1a载体上面含有一个EK蛋白酶切位点，当将目的基因使用载体上面的PshAI限制性内切酶位点插入进去时，可以通过EK蛋白酶将载体上的融合的所有氨基酸序列包括GST标签序列，全部切除掉。
pET-43.1EK/LIC载体序列
LOCUS pET-43.1 Ek/LIC 7295 bp DNA Circular SYN
DEFINITION pET-43.1 Ek/LIC
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
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
source 1..7295
/organism="pET-43.1 Ek/LIC"
/mol_type="other DNA"
misc_feature 69..87
/label="T7_Terminal_primer"
terminator complement(141..168)
/label="rrnB_T2_terminator"
terminator complement(300..343)
/label="rrnB_T1_terminator"
terminator complement(309..466)
/label="rrnB_terminator"
CDS complement(509..2338)
/label="ORF frame 2"
misc_feature complement(701..715)
/label="EK"
misc_feature complement(767..811)
/label="S15"
misc_feature complement(821..838)
/label="6xHIS"
CDS 824..2152
/label="ORF frame 2"
CDS 2339..3067
/label="ORF frame 2"
misc_feature complement(2347..2363)
/label="T7_transl_en_RBS"
misc_feature complement(2381..2408)
/label="lacO"
promoter complement(2408..2426)
/label="T7_promoter"
gene 2462..2725
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(2514..2533)
/label="pBRrevBam_primer"
misc_feature 2808..3899
/label="lacI"
CDS 2940..3899
/label="ORF frame 3"
misc_feature 4471..4662
/label="ROP"
misc_feature complement(4656..4678)
/label="pGEX_3_primer"
rep_origin complement(5077..5696)
/label="pBR322_origin"
promoter 5783..5811
/label="AmpR_promoter"
gene 5854..6714
/label="Ampicillin"
/gene="Ampicillin"
CDS 5854..6714
/label="ORF frame 1"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca
121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
661 gatccacgag ctcagatatc gttgaggaga agcccggact cttgtcgtcg tcatcaatcg
721 taccagaacc cgcgctgccc cgggggacca gaccagttgg cggtggcgag tccatgtgct
781 ggcgttcaaa tttcgcagca gcggtttctt tacccgcgga gtgatggtga tggtgatgac
841 cagaaccact agtcgcttcg tcaccgaacc agcaaatatt acgggcagcc ataatcagtg
901 ctccggcttt ttcgtcggtc aacccttcga tatcagccag atcatcaatg ccctgttcgg
961 cgagatcttc cagcgtacaa acgccacggg cggccagttt gaatgccaaa tcacgatcta
1021 ccccttcaag gttcagcaga tcgtcagccg gtttgttatc accgaggctt tcttcctggg
1081 cctgtgcaat ggtggccagt gcatttttag cacgctcgcg cagtgcttca acggtcggct
1141 catcaaggcc ttcgatttcc aacagctctt tcatcggcac ataggccaat tcttccagcg
1201 tcgagaagcc ttcttctacc agaacagtcg cgaagtcttc gtcgatgtcg agatatttgg
1261 tgaaggtgtc gatcgctgcg tgcgcttccg cctgatgctt agcttgcagg tcgtcaacgg
1321 tcatcacgtt gagttcccaa ccgctcagtt gcgaagccag acgcacgttc tgaccgttac
1381 ggccaatcgc ctgcgccaga ttaccggctt caacggcgat gtccatggtg tgtttatctt
1441 catccaccac gatagaagca acgtctgccg gtgccattgc gttaatcacg aactgcgccg
1501 ggttatcatc ccacaggacg atatcgatac gctcgccacc cagttcagta gacaccgcct
1561 gaacacgcgc gccacgcata cctacgcaag cacctaccgg atcgatacgt ttatcgttgg
1621 ttttcaccgc gattttcgca cgagaacccg gatcgcgagc cgctgcttta atttcaatca
1681 cttcttcgcc gatttctggc acttcaatac ggaacagttc gatcagcatt tccggcttgg
1741 aacgagtgac gaacagttgc gcgccacgcg cttccgggcg aacggaatag agcacgccac
1801 gaacgcggtc gccagggcgg aagttttcac gcggcagcat atcttcgcgc aggatcacgg
1861 cttcagcgtt gttgcccaga tccagagaga tgttgtcgcg gtttactttt ttcaccacgc
1921 cggtgatgat ttcaccttcg tgttcacgga actgatcaac caccatcgca cgttcggctt
1981 cacgcacttt ctgcacgata acctgttttg ccgtctgggt agtgatacgg tcaaaggtaa
2041 cagactcaat ctgatcttca acgtaatcgc ccaggttcag gctttcatct tcataacgtg
2101 cggcttcaag ggtgatttcc ttggtcggct gggtgacttc atcaacaact aaccagcgac
2161 ggaaagtgtc aaaatcaccg cttttgcgat cgatctgtac gcggacgtcg atctcttgtt
2221 catatttttt ctttgttgct gtcgccagcg cgctttccaa tgcttcgaaa atcttctcgc
2281 gaggtagcgc cttttcattg gatacggctt caactacagc caaaatttct ttgttcatat
2341 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
2401 caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta
2461 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
2521 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
2581 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
2641 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
2701 aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa
2761 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
2821 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
2881 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
2941 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
3001 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
3061 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
3121 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
3181 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
3241 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
3301 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
3361 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
3421 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
3481 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
3541 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
3601 tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
3661 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
3721 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
3781 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
3841 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
3901 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc
3961 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tagcatgatc
4021 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
4081 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
4141 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
4201 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
4261 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
4321 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
4381 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
4441 aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac
4501 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
4561 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
4621 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
4681 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
4741 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
4801 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatatgcg
4861 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
4921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
4981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
5041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
5101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
5161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
5221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
5281 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
5341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
5401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
5461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
5521 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
5581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
5641 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
5701 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaccta
5761 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
5821 aaatgcttca atcatgattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
5881 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
5941 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
6001 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
6061 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
6121 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
6181 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
6241 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
6301 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
6361 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
6421 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
6481 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
6541 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
6601 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
6661 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
6721 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
6781 atctaggtga agatcctttt tgataatctc atgccatcgt gccgccagaa attgtaaacg
6841 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
6901 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
6961 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
7021 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
7081 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
7141 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
7201 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
7261 ttaatgcgcc gctacagggc gcgtcccatt cgcca
//
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