pET-43.1a(+)

pET-43.1a(+)

pET43.1a载体基本信息

出品公司:	Ybscience
别名:	pET43.1a, pET 43.1a
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:	7275 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:	C term His tag only; N term thrombin cleavage site; N term enterokinase 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

pET43.1a载体质粒图谱和多克隆位点信息

Feature Name	Start	End
T7_Terminal_primer	68	86
rrnB_T2_terminator	167	140
rrnB_T1_terminator	342	299
rrnB_terminator	465	308
EK	694	680
S15	790	746
6xHIS	817	800
T7_transl_en_RBS	2342	2326
lacO	2387	2360
T7_promoter	2405	2387
tet (300 - 563)	2441	2704
pBRrevBam_primer	2512	2493
lacI	2787	3878
ROP	4450	4641
pGEX_3_primer	4657	4635
pBR322_origin	5675	5056
AmpR_promoter	5762	5790
Ampicillin	5833	6693

ORF	Start	End
ORF frame 3	2317	548
ORF frame 2	617	2131
ORF frame 2	2318	3046
ORF frame 3	2919	3878
ORF frame 1	5833	6693

Enzyme Name	Cut
PacI	502
XhoI	529
PmlI	575
NotI	591
EagI	591
HindIII	598
KpnI	623
SalI	625
PstI	638
AscI	640
EcoRI	653
BamHI	659
SacI	671
SmaI	720
XmaI	718
SacII	796
SpeI	827
BglII	942
MscI	1074
StuI	1127
NcoI	1402
NdeI	2316
XbaI	2354
ApaI	3357
HpaI	3652
FspI	6397

 

pET43.1a载体简介

 

The pET-43.1 series of vectors are designed for cloning and high-level expression of peptide sequences fused with the 491 aa Nus•Tag™ protein. 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.

 

 

pET43.1a载体序列

LOCUS       pET-43.1 a (+)    7275 bp     DNA  Circular    SYN
DEFINITION  pET-43.1 a (+)
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..7275
                     /organism="pET-43.1 a (+)"
                     /mol_type="other DNA"
     misc_feature    68..86
                     /label="T7_Terminal_primer"
     terminator      complement(140..167)
                     /label="rrnB_T2_terminator"
     terminator      complement(299..342)
                     /label="rrnB_T1_terminator"
     terminator      complement(308..465)
                     /label="rrnB_terminator"
     CDS             complement(548..2317)
                     /label="ORF frame 3"
     CDS             617..2131
                     /label="ORF frame 2"
     misc_feature    complement(680..694)
                     /label="EK"
     misc_feature    complement(746..790)
                     /label="S15"
     misc_feature    complement(800..817)
                     /label="6xHIS"
     CDS             2318..3046
                     /label="ORF frame 2"
     misc_feature    complement(2326..2342)
                     /label="T7_transl_en_RBS"
     misc_feature    complement(2360..2387)
                     /label="lacO"
     promoter        complement(2387..2405)
                     /label="T7_promoter" 
     gene            2441..2704
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(2493..2512)
                     /label="pBRrevBam_primer"
     misc_feature    2787..3878
                     /label="lacI"
     CDS             2919..3878
                     /label="ORF frame 3"
     misc_feature    4450..4641
                     /label="ROP" 
     misc_feature    complement(4635..4657)
                     /label="pGEX_3_primer"
     rep_origin      complement(5056..5675)
                     /label="pBR322_origin"
     promoter        5762..5790
                     /label="AmpR_promoter"
     gene            5833..6693
                     /label="Ampicillin"
                     /gene="Ampicillin" 
     CDS             5833..6693
                     /label="ORF frame 1"   
ORIGIN
    1 tccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag
   61 gggttatgct agttattgct cagcggatac atatttgaat gtatttagaa aaataaacaa
  121 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct taatttgatg
  181 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt
  241 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
  301 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggcagttcc
  361 ctactctcgc atggggagac cccacactac catcggcgct acggcgtttc acttctgagt
  421 tcggcatggg gtcaggtggg accaccgcgc tactgccgcc aggcaaattc tgttttatca
  481 gcctaggaac gcccaactta attaacatta gtggtggtgg tggtggtgct cgagatcctc
  541 tgggtcttca ggagcgagtt ctggctggct tgcacgtgta tacagctgtg cggccgcaag
  601 cttcgaacgc gtatcgatgg taccgtcgac gtcctgcagg cgcgcctgta cagaattcgg
  661 atccacgagc tcccggactc 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 tgttcatatg tatatctcct tcttaaagtt
 2341 aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg
 2401 tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca tcgtggccgg
 2461 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
 2521 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg
 2581 ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc
 2641 ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg
 2701 agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat
 2761 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
 2821 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
 2881 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
 2941 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
 3001 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
 3061 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
 3121 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
 3181 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
 3241 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
 3301 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
 3361 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
 3421 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
 3481 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
 3541 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggacatct
 3601 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
 3661 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
 3721 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
 3781 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
 3841 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt
 3901 tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt caacccagtc
 3961 agctccttcc ggtgggcgcg gggcatgact agcatgatcg tgctcctgtc gttgaggacc
 4021 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
 4081 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
 4141 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
 4201 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
 4261 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
 4321 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
 4381 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
 4441 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
 4501 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
 4561 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
 4621 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
 4681 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
 4741 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
 4801 catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac cgcacagatg
 4861 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
 4921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
 4981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
 5041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
 5101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
 5161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
 5221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
 5281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
 5341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
 5401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
 5461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
 5521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
 5581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
 5641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
 5701 gaacgaaaac tcacgttaag ggattttggt catgacctat ttgtttattt ttctaaatac
 5761 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa tcatgattga
 5821 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
 5881 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
 5941 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
 6001 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
 6061 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
 6121 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
 6181 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
 6241 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
 6301 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
 6361 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
 6421 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
 6481 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
 6541 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
 6601 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
 6661 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
 6721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
 6781 gataatctca tgccatcgtg ccgccagaaa ttgtaaacgt taatattttg ttaaaattcg
 6841 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
 6901 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
 6961 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
 7021 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
 7081 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
 7141 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
 7201 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
 7261 cgtcccattc gccaa
//