pETite N-His Kan

Vector for Expresso® cloning to add an N-terminal 6xHis tag, with inducible expression from the T7 promoter.

Sequence Author: Lucigen

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HomePlasmidsLucigen VectorspETite N-His Kan
XbaI (2221) DraI - SwaI (2198) tonB terminator AvaI - BsoBI - PaeR7I - PspXI - XhoI (2014) NruI (1959) EarI (1800) TsoI (1777) PasI (1741) EcoNI (1703) SspI (1691) BsrFI (1658) AsiSI - PvuI (1618) Bpu10I - BsmBI - Esp3I (1596) TaqII (1364) PflMI (1356) XmnI (1113) BanI (45) pETite T7 Forward (T7 Promoter) (51 .. 70) T7 promoter lac operator PmlI (92) NdeI (141) ATG EaeI - EagI - NotI (172) BlpI (184) StyI (206) pETite Reverse (213 .. 232) T3Te terminator AcuI (492) AlwNI (625) ApaLI (720) BsiHKAI (724) BseYI (730) PspFI (734) HaeII (794) BciVI (836) BssSI - BssSαI (861) PspOMI (1032) ApaI (1036) PvuII (1080) pETite N-His Kan 2235 bp
XbaI  (2221)
1 site		 T C T A G A A G A T C T
DraI  (2198)
1 site		 T T T A A A A A A T T T
SwaI  (2198)
1 site		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
AvaI  (2014)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2014)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (2014)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2014)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2014)
1 site		 C T C G A G G A G C T C
NruI  (1959)
1 site		 T C G C G A A G C G C T
EarI  (1800)
1 site		 C T C T T C N G A G A A G N N N N
Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
TsoI  (1777)
1 site		 T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9
Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PasI  (1741)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
EcoNI  (1703)
1 site		 C C T N N N N N A G G G G A N N N N N T C C
The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
SspI  (1691)
1 site		 A A T A T T T T A T A A
BsrFI  (1658)
1 site		 R C C G G Y Y G G C C R
Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
AsiSI  (1618)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (1618)
1 site		 C G A T C G G C T A G C
Bpu10I  (1596)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BsmBI  (1596)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (1596)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
TaqII  (1364)
1 site		 G A C C G A ( N ) 9 N N C T G G C T ( N ) 9
Sticky ends from different TaqII sites may not be compatible.
PflMI  (1356)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
XmnI  (1113)
1 site		 G A A N N N N T T C C T T N N N N A A G
BanI  (45)
1 site		 G G Y R C C C C R Y G G
Sticky ends from different BanI sites may not be compatible.
PmlI  (92)
1 site		 C A C G T G G T G C A C
NdeI  (141)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
EaeI  (172)
1 site		 Y G G C C R R C C G G Y
EagI  (172)
1 site		 C G G C C G G C C G G C
NotI  (172)
1 site		 G C G G C C G C C G C C G G C G
BlpI  (184)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
StyI  (206)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
AcuI  (492)
1 site		 C T G A A G ( N ) 14 N N G A C T T C ( N ) 14
Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (625)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
ApaLI  (720)
1 site		 G T G C A C C A C G T G
BsiHKAI  (724)
1 site		 G W G C W C C W C G W G
Sticky ends from different BsiHKAI sites may not be compatible.
BseYI  (730)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PspFI  (734)
1 site		 C C C A G C G G G T C G
HaeII  (794)
1 site		 R G C G C Y Y C G C G R
BciVI  (836)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
BssSI  (861)
1 site		 C A C G A G G T G C T C
BssSαI  (861)
1 site		 C A C G A G G T G C T C
PspOMI  (1032)
1 site		 G G G C C C C C C G G G
ApaI  (1036)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PvuII  (1080)
1 site		 C A G C T G G T C G A C
pETite T7 Forward (T7 Promoter)
20-mer  /  40% GC
1 binding site
51 .. 70  =  20 annealed bases
Tm  =  50°C
pETite Reverse
20-mer  /  55% GC
1 binding site
213 .. 232  =  20 annealed bases
Tm  =  56°C
KanR
1233 .. 2048  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
1233 .. 2048  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
391 .. 978  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
391 .. 978  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
1041 .. 1232  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
1041 .. 1232  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
cat promoter
2049 .. 2139  =  91 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
cat promoter
2049 .. 2139  =  91 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
T7 terminator
195 .. 242  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
195 .. 242  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
tonB terminator
2140 .. 2171  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
tonB terminator
2140 .. 2171  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
T3Te terminator
340 .. 369  =  30 bp
phage T3 early transcription terminator
T3Te terminator
340 .. 369  =  30 bp
phage T3 early transcription terminator
T7Te terminator
990 .. 1017  =  28 bp
phage T7 early transcription terminator
T7Te terminator
990 .. 1017  =  28 bp
phage T7 early transcription terminator
RBS
113 .. 135  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
113 .. 135  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
ATG
143 .. 145  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
143 .. 145  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
146 .. 163  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
146 .. 163  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
T7 promoter
51 .. 69  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
51 .. 69  =  19 bp
promoter for bacteriophage T7 RNA polymerase
lac operator
71 .. 86  =  16 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
71 .. 86  =  16 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
ORF:  948 .. 1175  =  228 bp
ORF:  75 amino acids  =  8.6 kDa
ORF:  1233 .. 2048  =  816 bp
ORF:  271 amino acids  =  30.9 kDa
ORF:  653 .. 1138  =  486 bp
ORF:  161 amino acids  =  18.0 kDa
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Individual Sequences & Maps

pAviTag C-His Kan
pEZ BAC
pRANGER-BTB-2
pSMART GC HK (linearized)
pAviTag N-His Kan
pEZ BAC (BamHI linearized)
pRANGER-BTB-3
pSMART GC LK (linearized)
pAviTag-C N-His SUMO Kan
pEZ BAC (HpaI linearized)
pRANGER-BTB-5
pSMART HCAmp
pAviTag-N N-His SUMO Kan
pGC Blue (linearized)
pRham C-His Kan
pSMART HCAmp (linearized)
pETite C-His Kan
pInvRecA
pRham N-His Kan
pSMART HCKan
pETite N-His Kan
pJAZZ-OC
pRham N-His SUMO Kan
pSMART HCKan (linearized)
pETite N-His SUMO Kan
pJAZZ-OK
pSMART BAC v2.0
pSMART LCAmp
pEZSeq-Amp
pJW168
pSMART BAC v2.0 (BamHI linearized)
pSMART LCAmp (linearized)
pEZSeq-Amp (linearized)
pME-HA (linearized)
pSMART BAC v2.0 (EcoRI linearized)
pSMART LCKan
pEZSeq-Kan
pNGS FOS (linearized)
pSMART BAC v2.0 (HindIII linearized)
pSMART LCKan (linearized)
pEZSeq-Kan (linearized)
pRANGER-BTB-1
pSMART FOS v2.0 (linearized)

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