pET-23(+)

pET-23(+)

pET23载体基本信息

出品公司:	Ybscience
载体名称:	pET-23a
质粒类型:	大肠杆菌表达载体；蛋白纯化
高拷贝/低拷贝:	低拷贝
克隆方法:	多克隆位点；限制性内切酶
启动子:	T7/ lacI
载体大小:	3592 bp
5' 测序引物及序列:	T7 Fwd:  5'd[TAATACGACTCACTATAGGG]3'
3' 测序引物及序列:	--
载体标签:	His Tag （C-端）
载体抗性:	氨苄青霉素（Ampicillin）
克隆菌株:	DH5a
表达菌株:	BL21系列
备注:	pET-23a，b，c仅阅读框（读码框）不同，彼此相差1个bp。
稳定性:	稳表达
组成型/诱导型:	诱导型（IPTG)
病毒/非病毒:	非病毒

 

pET23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
T7_promoter	245	227
ROP	754	945
pGEX_3_primer	961	939
pBR322_origin	1979	1360
Ampicillin	2994	2134
AmpR_promoter	3064	3036

ORF	Start	End
ORF frame 2	2994	2134

Enzyme Name	Cut
XhoI	158
NotI	166
EagI	166
HindIII	173
SalI	179
SacI	190
EcoRI	192
BamHI	198
BglII	260
MscI	285
PvuII	905
PstI	2452

 

pET23载体简介

pET-23(+) (Cat. No. 69771-3) is a transcription vector designed for expression from bacterial translation signals carried within a cloned insert. It therefore lacks the ribosome binding site and ATG start codon present on the pET translation vectors. A C-terminal His•Tag® sequence is available. 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 (Cat. No. 69337-3). 
 

pET23载体序列

LOCUS       pET-23(+)    3592 bp     DNA    SYN
DEFINITION  pET-23(+)
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..3592
                     /organism="pET-23(+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     promoter        complement(227..245)
                     /label="T7_promoter"
     misc_feature    754..945
                     /label="ROP"
     misc_feature    complement(939..961)
                     /label="pGEX_3_primer"
     rep_origin      complement(1360..1979)
                     /label="pBR322_origin"
     gene            complement(2134..2994)
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             complement(2134..2994)
                     /label="ORF frame 2"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        complement(3036..3064)
                     /label="AmpR_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
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 tcctagaggg aaaccgttgt ggtctcccta tagtgagtcg
  241 tattaatttc gcgggatcga gatctcgggc agcgttgggt cctggccacg ggtgcgcatg
  301 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
  361 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
  421 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
  481 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
  541 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
  601 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
  661 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
  721 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
  781 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
  841 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
  901 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
  961 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
 1021 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
 1081 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat
 1141 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
 1201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
 1261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
 1321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
 1381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
 1441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
 1501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
 1561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
 1621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
 1681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
 1741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
 1801 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
 1861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
 1921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
 1981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
 2041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
 2101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
 2161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
 2221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
 2281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
 2341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
 2401 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt
 2461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
 2521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
 2581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
 2641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
 2701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
 2761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
 2821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
 2881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
 2941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
 3001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
 3061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
 3121 acctgaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
 3181 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
 3241 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
 3301 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
 3361 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
 3421 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
 3481 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
 3541 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca