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Plasmid Files

pTrcHis C

Bacterial vector for inducible expression of N-terminally 6xHis-tagged proteins. For other reading frames, use pTrcHis A or pTrcHis B.

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pTrcHis C Sequence and MappTrcHis C.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 PluTI (4284) SfoI (4282) NarI * (4281) KasI (4280) EcoRV (4091) ApaI - BanII (3852) PspOMI (3848) BstEII (3822) BclI * (3655) MluI (3641) PflMI (3223) NsiI (3204) SphI (3202) MauBI (3180) PfoI (3112) PflFI - Tth111I (3013) BsaAI (3007) BstZ17I (2988) AccI (2987) BspQI - SapI (2874) PciI (2757) AlwNI (2348) BfuAI - BspMI (72) trc promoter lac operator gene 10 translational enhancer RBS BtgI - NcoI - StyI (411) ATG 6xHis NheI (449) BmtI (453) T7 tag (gene 10 leader) Xpress™ tag BsaBI (504) BamHI (513) AvaI - BsoBI - PaeR7I - XhoI (522) BmeT110I (523) BglII (526) PstI (534) Acc65I (538) KpnI (542) EcoRI (550) BstBI (554) HindIII (557) rrnB T1 terminator rrnB T2 terminator ScaI (1388) PvuI (1500) FspI (1646) BglI (1751) AhdI (1869) pTrcHis C 4412 bp
PluTI  (4284)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (4282)
1 site
G G C G C C C C G C G G
NarI  (4281)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (4280)
1 site
G G C G C C C C G C G G
EcoRV  (4091)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
ApaI  (3852)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (3852)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (3848)
1 site
G G G C C C C C C G G G
BstEII  (3822)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BclI  (3655)
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.
MluI  (3641)
1 site
A C G C G T T G C G C A
PflMI  (3223)
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.
NsiI  (3204)
1 site
A T G C A T T A C G T A
SphI  (3202)
1 site
G C A T G C C G T A C G
MauBI  (3180)
1 site
C G C G C G C G G C G C G C G C
PfoI  (3112)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
PflFI  (3013)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3013)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsaAI  (3007)
1 site
Y A C G T R R T G C A Y
BstZ17I  (2988)
1 site
G T A T A C C A T A T G
AccI  (2987)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BspQI  (2874)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2874)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
PciI  (2757)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (2348)
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.
BfuAI  (72)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (72)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BtgI  (411)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (411)
1 site
C C A T G G G G T A C C
StyI  (411)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NheI  (449)
1 site
G C T A G C C G A T C G
BmtI  (453)
1 site
G C T A G C C G A T C G
BsaBI  (504)
1 site
G A T N N N N A T C C T A N N N N T A G
BamHI  (513)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
AvaI  (522)
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  (522)
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  (522)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (522)
1 site
C T C G A G G A G C T C
BmeT110I  (523)
1 site
C Y C G R G G R G C Y C
BglII  (526)
1 site
A G A T C T T C T A G A
PstI  (534)
1 site
C T G C A G G A C G T C
Acc65I  (538)
1 site
G G T A C C C C A T G G
KpnI  (542)
1 site
G G T A C C C C A T G G
EcoRI  (550)
1 site
G A A T T C C T T A A G
BstBI  (554)
1 site
T T C G A A A A G C T T
HindIII  (557)
1 site
A A G C T T T T C G A A
ScaI  (1388)
1 site
A G T A C T T C A T G A
PvuI  (1500)
1 site
C G A T C G G C T A G C
FspI  (1646)
1 site
T G C G C A A C G C G T
BglI  (1751)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
AhdI  (1869)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
lacI
3291 .. 4373  =  1083 bp
360 amino acids  =  38.5 kDa
Product: lac repressor
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).
lacI
3291 .. 4373  =  1083 bp
360 amino acids  =  38.5 kDa
Product: lac repressor
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).
AmpR
1082 .. 1942  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   1082 .. 1150  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1082 .. 1942  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   1151 .. 1942  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1082 .. 1942  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2113 .. 2701  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2113 .. 2701  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
990 .. 1081  =  92 bp
AmpR promoter
990 .. 1081  =  92 bp
rrnB T1 terminator
765 .. 851  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
765 .. 851  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
lacIq promoter
3213 .. 3290  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
lacIq promoter
3213 .. 3290  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
rrnG antiterminator
263 .. 332  =  70 bp
antiterminator from the E. coli rrnG leader region
(Berg et al., 1989)
rrnG antiterminator
263 .. 332  =  70 bp
antiterminator from the E. coli rrnG leader region
(Berg et al., 1989)
MCS
513 .. 562  =  50 bp
multiple cloning site
MCS
513 .. 562  =  50 bp
multiple cloning site
T7 tag (gene 10 leader)
446 .. 478  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene
10
promotes efficient translation in E. coli
T7 tag (gene 10 leader)
446 .. 478  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene
10
promotes efficient translation in E. coli
trc promoter
193 .. 222  =  30 bp
   Segment 1:  -35  
   193 .. 198  =  6 bp
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
trc promoter
193 .. 222  =  30 bp
   Segment 2:  
   199 .. 215  =  17 bp
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
trc promoter
193 .. 222  =  30 bp
   Segment 3:  -10  
   216 .. 222  =  7 bp
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
trc promoter
193 .. 222  =  30 bp
3 segments
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
rrnB T2 terminator
943 .. 970  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
943 .. 970  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
minicistron
383 .. 409  =  27 bp
8 amino acids  =  1.1 kDa
Product: synthetic cistron containing a ribosome
binding site (Shine-Dalgarno sequence), for
enhancing the bacterial expression of a downstream
cistron (Schoner, 1997)
This first cistron should terminate 3 bp upstream of
the ATG for the second cistron.
minicistron
383 .. 409  =  27 bp
8 amino acids  =  1.1 kDa
Product: synthetic cistron containing a ribosome
binding site (Shine-Dalgarno sequence), for
enhancing the bacterial expression of a downstream
cistron (Schoner, 1997)
This first cistron should terminate 3 bp upstream of
the ATG for the second cistron.
Xpress™ tag
482 .. 505  =  24 bp
8 amino acids  =  998.0 Da
   Segment 1:  
   482 .. 490  =  9 bp
   3 amino acids  =  409.5 Da
Product: Xpress™ epitope tag, including an
enterokinase recognition and cleavage site
Xpress™ tag
482 .. 505  =  24 bp
8 amino acids  =  998.0 Da
   Segment 2:  enterokinase site  
   491 .. 505  =  15 bp
   5 amino acids  =  606.5 Da
Product: Xpress™ epitope tag, including an
enterokinase recognition and cleavage site
Xpress™ tag
482 .. 505  =  24 bp
8 amino acids  =  998.0 Da
2 segments
Product: Xpress™ epitope tag, including an
enterokinase recognition and cleavage site
6xHis
425 .. 442  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
425 .. 442  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac operator
230 .. 246  =  17 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
230 .. 246  =  17 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).
gene 10 translational enhancer
346 .. 354  =  9 bp
translational enhancer from gene 10 of
bacteriophage T7 (Olins and Rangwala, 1989)
gene 10 translational enhancer
346 .. 354  =  9 bp
translational enhancer from gene 10 of
bacteriophage T7 (Olins and Rangwala, 1989)
RBS
369 .. 373  =  5 bp
ribosome binding site
RBS
369 .. 373  =  5 bp
ribosome binding site
ATG
413 .. 415  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
413 .. 415  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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