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Plasmid Files

pAviTag N-His Kan

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

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pAviTag N-His Kan Sequence and MappAviTag N-His Kan.dna
Map and Sequence File   
Sequence Author:  Lucigen
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 DraI - SwaI (2322) BsaAI (2253) cat promoter PaeR7I - PspXI - XhoI (2138) EarI (1924) TsoI (1901) PasI (1865) EcoNI (1827) SspI (1815) BsrFI (1782) AsiSI - PvuI (1742) Bpu10I - BsmBI (1720) AseI (1539) TaqII (1488) PflMI (1480) XmnI (1237) PvuII (1204) pRham Forward (101 .. 124) BsaI (116) NdeI (181) ATG BseRI (268) BsrBI (295) EaeI - EagI - NotI (296) BlpI (308) StyI (330) pETite Reverse (337 .. 356) AcuI (616) AlwNI (749) ApaLI (844) BseYI (854) PspFI (858) BciVI (960) BssS α I (985) PspOMI (1156) ApaI (1160) pAviTag N-His Kan 2358 bp
DraI  (2322)
1 site
T T T A A A A A A T T T
SwaI  (2322)
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  (2253)
1 site
Y A C G T R R T G C A Y
PaeR7I  (2138)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2138)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2138)
1 site
C T C G A G G A G C T C
EarI  (1924)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
TsoI  (1901)
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  (1865)
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  (1827)
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  (1815)
1 site
A A T A T T T T A T A A
BsrFI  (1782)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
AsiSI  (1742)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1742)
1 site
C G A T C G G C T A G C
Bpu10I  (1720)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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  (1720)
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.
AseI  (1539)
1 site
A T T A A T T A A T T A
TaqII  (1488)
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  (1480)
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  (1237)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuII  (1204)
1 site
C A G C T G G T C G A C
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.
BsrBI  (295)
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  (296)
1 site
Y G G C C R R C C G G Y
EagI  (296)
1 site
C G G C C G G C C G G C
NotI  (296)
1 site
G C G G C C G C C G C C G G C G
BlpI  (308)
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  (330)
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  (616)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
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  (749)
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  (844)
1 site
G T G C A C C A C G T G
BseYI  (854)
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  (858)
1 site
C C C A G C G G G T C G
BciVI  (960)
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.
BssSαI  (985)
1 site
C A C G A G G T G C T C
PspOMI  (1156)
1 site
G G G C C C C C C G G G
ApaI  (1160)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
pRham Forward
24-mer  /  50% GC
1 binding site
101 .. 124  =  24 annealed bases
Tm  =  58°C
pETite Reverse
20-mer  /  55% GC
1 binding site
337 .. 356  =  20 annealed bases
Tm  =  56°C
KanR
1357 .. 2172  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
1357 .. 2172  =  816 bp
271 amino acids  =  30.9 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
ori
515 .. 1102  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
515 .. 1102  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
rop
1165 .. 1356  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
1165 .. 1356  =  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
2173 .. 2263  =  91 bp
promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase
cat promoter
2173 .. 2263  =  91 bp
promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase
T7 terminator
319 .. 366  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
319 .. 366  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
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
tonB terminator
2264 .. 2295  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
tonB terminator
2264 .. 2295  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
T3Te terminator
464 .. 493  =  30 bp
phage T3 early transcription terminator
T3Te terminator
464 .. 493  =  30 bp
phage T3 early transcription terminator
T7Te terminator
1114 .. 1141  =  28 bp
phage T7 early transcription terminator
T7Te terminator
1114 .. 1141  =  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)
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
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
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