pCold III

Vector for cold shock-induced expression in E. coli of a protein with a translation enhancing element.

Sequence Author: TaKaRa

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: Basic Cloning Vectors | More Plasmid Sets
No matches
pCold-F (208 .. 226) TEE NdeI (272) Eco53kI (279) SacI (281) Acc65I (283) KpnI (287) AbsI - PaeR7I - PspXI - XhoI (289) BamHI (295) MCS EcoRI (301) HindIII (307) SalI (313) AccI (314) PstI (323) XbaI (325) BfuAI - BspMI (326) pCold-R (354 .. 373) pCold™ III 4377 bp
NdeI  (272)
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.
Eco53kI  (279)
1 site
G A G C T C C T C G A G
SacI  (281)
1 site
G A G C T C C T C G A G
Acc65I  (283)
1 site
G G T A C C C C A T G G
KpnI  (287)
1 site
G G T A C C C C A T G G
AbsI  (289)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (289)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (289)
1 site
V C T C G A G B B G A G C T C V
XhoI  (289)
1 site
C T C G A G G A G C T C
BamHI  (295)
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.
EcoRI  (301)
1 site
G A A T T C C T T A A G
HindIII  (307)
1 site
A A G C T T T T C G A A
SalI  (313)
1 site
G T C G A C C A G C T G
AccI  (314)
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.
PstI  (323)
1 site
C T G C A G G A C G T C
XbaI  (325)
1 site
T C T A G A A G A T C T
BfuAI  (326)
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  (326)
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.
pCold-F
19-mer  /  58% GC
1 binding site
208 .. 226  =  19 annealed bases
Tm  =  60°C
pCold-R
20-mer  /  50% GC
1 binding site
354 .. 373  =  20 annealed bases
Tm  =  56°C
lacI
3132 .. 4214  =  1083 bp
360 amino acids  =  38.6 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
3132 .. 4214  =  1083 bp
360 amino acids  =  38.6 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
1341 .. 2201  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1341 .. 1409  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1341 .. 2201  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1410 .. 2201  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1341 .. 2201  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2372 .. 2960  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2372 .. 2960  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
681 .. 1136  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
681 .. 1136  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
cspA 3'UTR
338 .. 482  =  145 bp
3'UTR of the E. coli cold shock protein cspA gene (Mitta et al., 1997)
cspA 3'UTR
338 .. 482  =  145 bp
3'UTR of the E. coli cold shock protein cspA gene (Mitta et al., 1997)
cspA 5'UTR
122 .. 255  =  134 bp
5'UTR of the E. coli cold shock protein cspA gene (Mitta et al., 1997)
cspA 5'UTR
122 .. 255  =  134 bp
5'UTR of the E. coli cold shock protein cspA gene (Mitta et al., 1997)
AmpR promoter
1236 .. 1340  =  105 bp
AmpR promoter
1236 .. 1340  =  105 bp
lacI promoter
4215 .. 4292  =  78 bp
lacI promoter
4215 .. 4292  =  78 bp
cspA promoter
15 .. 81  =  67 bp
promoter of the E. coli cold shock protein cspA gene (Mitta et al., 1997)
cspA promoter
15 .. 81  =  67 bp
promoter of the E. coli cold shock protein cspA gene (Mitta et al., 1997)
MCS
271 .. 330  =  60 bp
multiple cloning site
MCS
271 .. 330  =  60 bp
multiple cloning site
lac operator
84 .. 100  =  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
84 .. 100  =  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).
TEE
256 .. 270  =  15 bp
5 amino acids  =  627.8 Da
Product: translation enhancing element for E. coli (Qing et al., 2004)
TEE
256 .. 270  =  15 bp
5 amino acids  =  627.8 Da
Product: translation enhancing element for E. coli (Qing et al., 2004)
ORF:  2930 .. 3349  =  420 bp
ORF:  139 amino acids  =  15.3 kDa
ORF:  1341 .. 2201  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3112 .. 3375  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3964 .. 4329  =  366 bp
ORF:  121 amino acids  =  13.1 kDa
ORF:  3132 .. 4091  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  1805 .. 2071  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
Click here to try SnapGene

Download pCold III.dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps

The maps, notes, and annotations in the zip file on this page are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as ’’www.snapgene.com/resources’’. Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Discover the most user-friendly molecular biology experience.