pAviTag-N N-His SUMO Kan

Vector for Expresso® cloning to add N-terminal 6xHis, biotinylation, and SUMO tags, with rhamnose-inducible expression.

Sequence Author: Lucigen

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HomePlasmidsLucigen VectorspAviTag-N N-His SUMO Kan
SwaI (2622) tonB terminator BsaAI (2553) PaeR7I - PspXI - XhoI (2438) EarI (2224) TsoI (2201) PasI (2165) EcoNI (2127) SspI (2115) BsrFI (2082) AsiSI - PvuI (2042) Bpu10I - BsmBI - Esp3I (2020) AseI (1839) TaqII (1788) PflMI (1780) XmnI (1537) PvuII (1504) ApaI (1460) PspOMI (1456) BsaI (116) RBS NdeI (181) ATG 6xHis AviTag™ BseRI (268) KflI - PpuMI (289) PstI - SbfI (298) DrdI (306) BglII (394) AclI (483) SUMO Forward (507 .. 530) BclI * (515) BsrBI (595) EaeI - EagI - NotI (596) StyI (630) pETite Reverse (637 .. 656) AlwNI (1049) ApaLI (1144) BseYI (1154) PspFI (1158) BciVI (1260) BssSI - BssSαI (1285) pAviTag-N N-His SUMO Kan 2659 bp
SwaI  (2622)
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.
BsaAI  (2553)
1 site		 Y A C G T R R T G C A Y
PaeR7I  (2438)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2438)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2438)
1 site		 C T C G A G G A G C T C
EarI  (2224)
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  (2201)
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  (2165)
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  (2127)
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  (2115)
1 site		 A A T A T T T T A T A A
BsrFI  (2082)
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  (2042)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (2042)
1 site		 C G A T C G G C T A G C
Bpu10I  (2020)
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  (2020)
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  (2020)
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.
AseI  (1839)
1 site		 A T T A A T T A A T T A
TaqII  (1788)
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  (1780)
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  (1537)
1 site		 G A A N N N N T T C C T T N N N N A A G
PvuII  (1504)
1 site		 C A G C T G G T C G A C
ApaI  (1460)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (1456)
1 site		 G G G C C C C C C G G G
BsaI  (116)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
NdeI  (181)
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.
BseRI  (268)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
KflI  (289)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different KflI sites may not be compatible.
PpuMI  (289)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
PstI  (298)
1 site		 C T G C A G G A C G T C
SbfI  (298)
1 site		 C C T G C A G G G G A C G T C C
DrdI  (306)
1 site		 G A C N N N N N N G T C C T G N N N N N N C A G
Sticky ends from different DrdI sites may not be compatible.
BglII  (394)
1 site		 A G A T C T T C T A G A
AclI  (483)
1 site		 A A C G T T T T G C A A
BclI  (515)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BsrBI  (595)
1 site		 C C G C T C G G C G A G
This recognition sequence is asymmetric, so ligating blunt ends generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures up to 50°C.
EaeI  (596)
1 site		 Y G G C C R R C C G G Y
EagI  (596)
1 site		 C G G C C G G C C G G C
NotI  (596)
1 site		 G C G G C C G C C G C C G G C G
StyI  (630)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
AlwNI  (1049)
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  (1144)
1 site		 G T G C A C C A C G T G
BseYI  (1154)
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  (1158)
1 site		 C C C A G C G G G T C G
BciVI  (1260)
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  (1285)
1 site		 C A C G A G G T G C T C
BssSαI  (1285)
1 site		 C A C G A G G T G C T C
SUMO Forward
24-mer  /  46% GC
1 binding site
507 .. 530  =  24 annealed bases
Tm  =  59°C
pETite Reverse
20-mer  /  55% GC
1 binding site
637 .. 656  =  20 annealed bases
Tm  =  56°C
KanR
1657 .. 2472  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
1657 .. 2472  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
815 .. 1402  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
815 .. 1402  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ATG
183 .. 185  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
183 .. 185  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
186 .. 203  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
186 .. 203  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
AviTag™
204 .. 248  =  45 bp
15 amino acids  =  1.8 kDa
Product: peptide tag that allows for enzymatic biotinylation
AviTag™
204 .. 248  =  45 bp
15 amino acids  =  1.8 kDa
Product: peptide tag that allows for enzymatic biotinylation
SUMO
300 .. 587  =  288 bp
96 amino acids  =  11.0 kDa
Product: ubiquitin-like protein tag
modified by Lucigen to prevent cleavage by eukaryotic SUMO proteases
SUMO
300 .. 587  =  288 bp
96 amino acids  =  11.0 kDa
Product: ubiquitin-like protein tag
modified by Lucigen to prevent cleavage by eukaryotic SUMO proteases
rop
1465 .. 1656  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
1465 .. 1656  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rhaB promoter
1 .. 119  =  119 bp
promoter of the E. coli rhaBAD operon, conferring tight induction with L-rhamnose and repression with D-glucose in the presence of RhaR and RhaS (Giacalone et al., 2006)
rhaB promoter
1 .. 119  =  119 bp
promoter of the E. coli rhaBAD operon, conferring tight induction with L-rhamnose and repression with D-glucose in the presence of RhaR and RhaS (Giacalone et al., 2006)
cat promoter
2473 .. 2563  =  91 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
cat promoter
2473 .. 2563  =  91 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
T7 terminator
619 .. 666  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
619 .. 666  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
tonB terminator
2564 .. 2595  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
tonB terminator
2564 .. 2595  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
T3Te terminator
764 .. 793  =  30 bp
phage T3 early transcription terminator
T3Te terminator
764 .. 793  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1414 .. 1441  =  28 bp
phage T7 early transcription terminator
T7Te terminator
1414 .. 1441  =  28 bp
phage T7 early transcription terminator
RBS
153 .. 175  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
153 .. 175  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
ORF:  1372 .. 1599  =  228 bp
ORF:  75 amino acids  =  8.6 kDa
ORF:  183 .. 590  =  408 bp
ORF:  135 amino acids  =  15.4 kDa
ORF:  1657 .. 2472  =  816 bp
ORF:  271 amino acids  =  30.9 kDa
ORF:  1077 .. 1562  =  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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