pET-44c(+)

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相关技术服务:质粒构建    基因合成    质粒大提

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pET44c载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET44c, pET 44c
质粒类型: 大肠杆菌蛋白表达
表达水平:
克隆方法: 多克隆位点,限制性内切酶
载体大小: 7309 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:
Both Nterm and Cterm His tags; Nterm thrombin cleavage site; Nterm enterokinase cleavage site; a,b,c vary by MCS.
产品目录号: 71124-3
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

pET44c载体图谱质粒图谱和多克隆位点信息

pet44c载体图谱
pet44c多克隆位点

Feature Name Start End  
T7_Terminal_primer 69 87
rrnB_T2_terminator 168 141
rrnB_T1_terminator 343 300
rrnB_terminator 466 309
EK 693 679
S15 789 745
6xHIS 2340 2323
T7_transl_en_RBS 2377 2361
lacO 2422 2395
T7_promoter 2440 2422
tet (300 - 563) 2476 2739
pBRrevBam_primer 2547 2528
lacI 2822 3913
ROP 4485 4676
pGEX_3_primer 4692 4670
pBR322_origin 5710 5091
AmpR_promoter 5797 5825
Ampicillin 5868 6728
ORF Start End  
ORF frame 1 427 2130
ORF frame 2 2352 499
ORF frame 1 2323 3081
ORF frame 2 2954 3913
ORF frame 3 5868 6728
Enzyme Name Cut
PacI 503
XhoI 530
PmlI 576
NotI 592
EagI 592
HindIII 599
KpnI 624
SalI 626
PstI 639
AscI 641
EcoRI 654
BamHI 660
SacI 672
SmaI 719
XmaI 717
SacII 795
SpeI 826
BglII 941
MscI 1073
StuI 1126
NcoI 1401
NdeI 2351
XbaI 2389
ApaI 3392
HpaI 3687
FspI 6432

pET44c载体简介

pET44系列载体是设计用来克隆和高水平表达含有495个氨基酸的Nus标签的蛋白表达载体。与pET-43.1系列载体相比,pET44系列载体额外编码一个N端的His蛋白纯化标签。 pET42a-c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。对pET44a载体的单链测序可以使用COLIDOWN primer(Cat. No. 70845-3).< 把表达的基因插入到载体的PshAI或者SmaI限制性内切酶位点时,可以通过EK蛋白酶或者Thrombin蛋白酶将融合表达的Nus标签完全切除。



pET Vectors 选择指南

pET44c载体序列

LOCUS       pET-44c(+)    7309 bp     DNA  Circular   SYN
DEFINITION  pET-44c(+)
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..7309
                     /organism="pET-44 c(+)"
                     /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             427..2130
                     /label="ORF frame 1"
     CDS             complement(499..2352)
                     /label="ORF frame 2"  
     misc_feature    complement(679..693)
                     /label="EK" 
     misc_feature    complement(745..789)
                     /label="S15"
     CDS             2323..3081
                     /label="ORF frame 1"       
     misc_feature    complement(2323..2340)
                     /label="6xHIS"  
     misc_feature    complement(2361..2377)
                     /label="T7_transl_en_RBS"   
     misc_feature    complement(2395..2422)
                     /label="lacO"      
     promoter        complement(2422..2440)
                     /label="T7_promoter"     
     gene            2476..2739
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"      
     misc_feature    complement(2528..2547)
                     /label="pBRrevBam_primer"  
     misc_feature    2822..3913
                     /label="lacI"  
     CDS             2954..3913
                     /label="ORF frame 2"  
     misc_feature    4485..4676
                     /label="ROP"    
     misc_feature    complement(4670..4692)
                     /label="pGEX_3_primer"   
     rep_origin      complement(5091..5710)
                     /label="pBR322_origin"   
     promoter        5797..5825
                     /label="AmpR_promoter"
     gene            5868..6728
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             5868..6728
                     /label="ORF frame 3"       
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 ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg
  721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc
  781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc
  841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa
  901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac
  961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc
 1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc
 1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa
 1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag
 1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg
 1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc
 1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt
 1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac
 1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat
 1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc
 1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg
 1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac
 1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc
 1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa
 1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc
 1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg
 1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac
 1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac
 2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt
 2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct
 2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt
 2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga
 2281 tacggcttca actacagcca aaatttcttt gttcatacta gaatggtgat ggtggtgatg
 2341 gctgctgccc atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
 2401 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg
 2461 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
 2521 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
 2581 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
 2641 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
 2701 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
 2761 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
 2821 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
 2881 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
 2941 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
 3001 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
 3061 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
 3121 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
 3181 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
 3241 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
 3301 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
 3361 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
 3421 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
 3481 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
 3541 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
 3601 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga
 3661 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
 3721 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
 3781 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
 3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
 3901 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
 3961 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
 4021 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
 4081 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
 4141 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
 4201 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
 4261 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
 4321 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
 4381 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
 4441 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
 4501 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
 4561 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
 4621 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
 4681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
 4741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
 4801 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
 4861 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
 4921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
 4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
 5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
 5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
 5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
 5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
 5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
 5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
 5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
 5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
 5521 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
 5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
 5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
 5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
 5761 ttggtcatga cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
 5821 acaataaccc tgataaatgc ttcaatcatg attgaaaaag gaagagtatg agtattcaac
 5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
 5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
 6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
 6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
 6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
 6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
 6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
 6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
 6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
 6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
 6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
 6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
 6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
 6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
 6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
 6781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgcca tcgtgccgcc
 6841 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
 6901 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
 6961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
 7021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
 7081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
 7141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
 7201 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
 7261 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
//

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