pET-44b(+)

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联系方式:I47-825O-882O

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pET44b载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET44b, pET 44b
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: 7310 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
表达宿主菌: BL21(DE3) Rosetta2(DE3)C43(DE3)
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: 71123-3
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

pET44b载体质粒图谱和多克隆位点信息

pet44b载体图谱
pet44b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 694 680
S15 790 746
6xHIS 2341 2324
T7_transl_en_RBS 2378 2362
lacO 2423 2396
T7_promoter 2441 2423
tet (300 - 563) 2477 2740
pBRrevBam_primer 2548 2529
lacI 2823 3914
ROP 4486 4677
pGEX_3_primer 4693 4671
pBR322_origin 5711 5092
AmpR_promoter 5798 5826
Ampicillin 5869 6729
ORF Start End  
ORF frame 2 2353 509
ORF frame 2 803 2131
ORF frame 2 2324 3082
ORF frame 3 2955 3914
ORF frame 1 5869 6729
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 720
XmaI 718
SacII 796
SpeI 827
BglII 942
MscI 1074
StuI 1127
NcoI 1402
NdeI 2352
XbaI 2390
ApaI 3393
HpaI 3688
FspI 6433

pET44b载体简介

The pET-44 vectors are designed for cloning and high-level expression of peptide sequences fused with the 495 aa Nus•Tag™ protein. Compared to the pET-43.1 series, the pET-44 vectors encode an additional N-terminal His•Tag. 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 circle 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 COLIDOWN primer (Cat. No. 70845-3). Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites (as shown below) by cleaving the Nus•Tag fusion protein with enterokinase or thrombin, respectively.

pET44b载体序列

LOCUS       pET-44 b(+)    7310 bp     DNA  Circular   SYN
DEFINITION  pET-44 b(+)
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..7310
                     /organism="pET-44 b(+)"
                     /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..2353)
                     /label="ORF frame 2"
     misc_feature    complement(680..694)
                     /label="EK"     
     misc_feature    complement(746..790)
                     /label="S15"  
     CDS             803..2131
                     /label="ORF frame 2"      
     CDS             2324..3082
                     /label="ORF frame 2"  
     misc_feature    complement(2324..2341)
                     /label="6xHIS"  
     misc_feature    complement(2362..2378)
                     /label="T7_transl_en_RBS" 
     misc_feature    complement(2396..2423)
                     /label="lacO"    
     promoter        complement(2423..2441)
                     /label="T7_promoter"    
     gene            2477..2740
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(2529..2548)
                     /label="pBRrevBam_primer"  
     misc_feature    2823..3914
                     /label="lacI" 
     CDS             2955..3914
                     /label="ORF frame 3"   
     misc_feature    4486..4677
                     /label="ROP"  
     misc_feature    complement(4671..4693)
                     /label="pGEX_3_primer"  
     rep_origin      complement(5092..5711)
                     /label="pBR322_origin"  
     promoter        5798..5826
                     /label="AmpR_promoter" 
     gene            5869..6729
                     /label="Ampicillin"
                     /gene="Ampicillin"    
     CDS             5869..6729
                     /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 ctccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc
  721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag
  781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt
  841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca
  901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa
  961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat
 1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg
 1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca
 1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca
 1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt
 1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac
 1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat
 1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa
 1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga
 1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac
 1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac
 1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca
 1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg
 1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga
 1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat
 1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt
 1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa
 1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa
 2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct
 2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc
 2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg
 2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg
 2281 atacggcttc aactacagcc aaaatttctt tgttcatact agaatggtga tggtggtgat
 2341 ggctgctgcc catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
 2401 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
 2461 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
 2521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
 2581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
 2641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
 2701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
 2761 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
 2821 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
 2881 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
 2941 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
 3001 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
 3061 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
 3121 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
 3181 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
 3241 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
 3301 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
 3361 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
 3421 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
 3481 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
 3541 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
 3601 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
 3661 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
 3721 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
 3781 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
 3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
 3901 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
 3961 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
 4021 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
 4081 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
 4141 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
 4201 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
 4261 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
 4321 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
 4381 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
 4441 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
 4501 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
 4561 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
 4621 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
 4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
 4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
 4801 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
 4861 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
 4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
 4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
 5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
 5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
 5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
 5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
 5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
 5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
 5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
 5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
 5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
 5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
 5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
 5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
 5761 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
 5821 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa
 5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
 5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
 6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
 6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
 6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
 6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
 6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
 6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
 6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
 6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
 6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
 6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
 6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
 6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
 6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
 6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc
 6841 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
 6901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
 6961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
 7021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
 7081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
 7141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
 7201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
 7261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//

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