pCold I

Vector for cold shock-induced expression in E. coli of a protein with a translation enhancing element followed by a 6xHis tag and a Factor Xa site.

Sequence Author: TaKaRa

Try SnapGene for Free
|Download SnapGene Viewer
Explore Over 2.7k Plasmids: Basic Cloning Vectors | More Plasmid Sets
No matches
HomePlasmidsBasic Cloning VectorspCold I
pCold-F (208 .. 226) TEE 6xHis Factor Xa site NdeI (302) Eco53kI (309) SacI (311) Acc65I (313) KpnI (317) AbsI - PaeR7I - PspXI - XhoI (319) BamHI (325) MCS EcoRI (331) HindIII (337) SalI (343) AccI (344) PstI (353) XbaI (355) BfuAI - BspMI (356) pCold-R (384 .. 403) pCold™ I 4407 bp
NdeI  (302)
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  (309)
1 site		 G A G C T C C T C G A G
SacI  (311)
1 site		 G A G C T C C T C G A G
Acc65I  (313)
1 site		 G G T A C C C C A T G G
KpnI  (317)
1 site		 G G T A C C C C A T G G
AbsI  (319)
1 site		 C C T C G A G G G G A G C T C C
PaeR7I  (319)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (319)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (319)
1 site		 C T C G A G G A G C T C
BamHI  (325)
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  (331)
1 site		 G A A T T C C T T A A G
HindIII  (337)
1 site		 A A G C T T T T C G A A
SalI  (343)
1 site		 G T C G A C C A G C T G
AccI  (344)
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  (353)
1 site		 C T G C A G G A C G T C
XbaI  (355)
1 site		 T C T A G A A G A T C T
BfuAI  (356)
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  (356)
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
384 .. 403  =  20 annealed bases
Tm  =  56°C
lacI
3162 .. 4244  =  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
3162 .. 4244  =  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
1371 .. 2231  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1371 .. 1439  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1371 .. 2231  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1440 .. 2231  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1371 .. 2231  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2402 .. 2990  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2402 .. 2990  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
711 .. 1166  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
711 .. 1166  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
cspA 3'UTR
368 .. 512  =  145 bp
3'UTR of the E. coli cold shock protein cspA gene (Mitta et al., 1997)
cspA 3'UTR
368 .. 512  =  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
1266 .. 1370  =  105 bp
AmpR promoter
1266 .. 1370  =  105 bp
lacI promoter
4245 .. 4322  =  78 bp
lacI promoter
4245 .. 4322  =  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
301 .. 360  =  60 bp
multiple cloning site
MCS
301 .. 360  =  60 bp
multiple cloning site
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)
6xHis
271 .. 288  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
271 .. 288  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
Factor Xa site
289 .. 300  =  12 bp
4 amino acids  =  473.5 Da
Product: Factor Xa recognition and cleavage site
Factor Xa site
289 .. 300  =  12 bp
4 amino acids  =  473.5 Da
Product: Factor Xa recognition and cleavage 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).
ORF:  2960 .. 3379  =  420 bp
ORF:  139 amino acids  =  15.3 kDa
ORF:  1371 .. 2231  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3142 .. 3405  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3994 .. 4359  =  366 bp
ORF:  121 amino acids  =  13.1 kDa
ORF:  3162 .. 4121  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  1835 .. 2101  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
Click here to try SnapGene

Download pCold I.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
Download SnapGene

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
Download SnapGene Viewer

Individual Sequences & Maps

BlueScribe
pEASY-T1 (linearized)
pJET1.2
pSMART LCAmp
CAG promoter
pEASY-T1 Simple (linearized)
pJET1.2 blunt
pSMART LCAmp (linearized)
chicken beta-actin promoter
pEASY-T3 (linearized)
pJW168
pSMART LCKan
CLIP-tag
pEASY-T5 Zero (linearized)
pKF 18k-2
pSMART LCKan (linearized)
CMV promoter
pEX-1
pKF 19k-2
pSP64 Poly(A)
GAL1 promoter
pEX-A
pKOV
pSP72
GST
pEX-C-GST
pLATE11 (linearized)
pSP73
HaloTag
pEX-C-His
pLATE31 (linearized)
pSpark Done (linearized)
HSV TK promoter
pEX-K
pLATE51 (linearized)
pSpark I (linearized)
lac operator
pEX-N-GST
pLATE52 (linearized)
pSpark II (linearized)
lac promoter
pEX-N-His
pLEXX-AK
pSpark III (linearized)
lac UV5 promoter
pEX-N-His-GST
pLPS-B (linearized)
pSpark IV (linearized)
lacZ
pEX1
pMAL-c5E
pSpark V (linearized)
lambda
pEX2
pMAL-c5G
pSpot1
LITMUS28
pEX3
pMAL-c5X
pSpot2
LITMUS28i
pEZSeq-Amp
pMAL-c5X-His
pSpot3
LITMUS29
pEZSeq-Amp (linearized)
pMAL-c6T
pSpot4
LITMUS38
pEZSeq-Kan
pMAL-p5E
pSTBlue-1
LITMUS38i
pEZSeq-Kan (linearized)
pMAL-p5G
pSTV28
LITMUS39
pEZ BAC
pMAL-p5X
pSTV29
M13mp18
pEZ BAC (BamHI linearized)
pMCS5
pT7Blue
M13mp19
pEZ BAC (HpaI linearized)
pMiniT
pT7Blue-2
MBP
pF1A T7
pMiniT (linearized)
pT7Blue-3
pACYC177
pF1K T7
pMiniT 2.0
pT7CFE1-CFtag
pACYC184
pF25A ICE T7
pMiniT 2.0 (linearized)
pT7CFE1-CGFP-HA-His
pAN7-1
pF25K ICE T7
pMMB190
pT7CFE1-CGST-HA-His
pANT7_cGST
pF3A WG (BYDV)
pMMB206
pT7CFE1-CHA
pANT7_nHA
pF3K WG (BYDV)
pMMB207
pT7CFE1-CHis
pAUR316
pFC20A HaloTag T7 SP6
pMMB208
pT7CFE1-CMyc
pBC KS(+)
pFC20K HaloTag T7 SP6
pMMB66EH
pT7CFE1-NFtag
pBC KS(-)
pFC30A His6HaloTag T7
pMMB66HE
pT7CFE1-NHA
pBC SK(+)
pFC30K His6HaloTag T7
pMMB67EH
pT7CFE1-NHA-CHis
pBC SK(-)
pFC7A (HQ)
pMMB67HE
pT7CFE1-NHis
pBeloBAC11
pFC7K (HQ)
pNEB193
pT7CFE1-NHis-GST
pBluescript II KS(+)
pFN18A HaloTag T7
pNEB206A (linearized)
pT7CFE1-NHis-GST-CHA
pBluescript II KS(-)
pFN18K HaloTag T7
pOSIP-CH
pT7CFE1-NMyc
pBluescript II SK(+)
pFN19A HaloTag T7 SP6
pOSIP-CO
pTOP Blunt V2 (linearized)
pBluescript II SK(-)
pFN19K HaloTag T7 SP6
pOSIP-CT
pTrcHis A
pBluescript KS(+)
pFN29A His6HaloTag T7
pOSIP-KC
pTrcHis B
pBluescript KS(-)
pFN29K His6HaloTag T7
pOSIP-KH
pTrcHis C
pBluescript SK(+)
pFN2A (GST)
pOSIP-KL
pTrcHis2 A
pBluescript SK(-)
pFN2K (GST)
pOSIP-KO
pTrcHis2 B
pBR322
pFN6A (HQ)
pOSIP-KP
pTrcHis2 C
pBS KS(+)
pFN6K (HQ)
pOSIP-KT
pTV 118N
pBS KS(-)
pGEM-11Zf(+)
pOSIP-TH
pTWV228
pBS SK(+)
pGEM-11Zf(-)
pOSIP-TO
pTXB1
pBS SK(-)
pGEM-3Z
pOSIP-TT
pTYB21
pBS(+)
pGEM-3Zf(+)
pPCR-Script Amp SK(+)
pType IIs
pBS(-)
pGEM-3Zf(-)
pPCR-Script Cam SK(+)
pTZ19R
pBSK(+) Simple-Amp
pGEM-4Z
pPTR I
pTZ19U
pBSK(+) Simple-Kan
pGEM-5Zf(+)
pPTR II
pTZ57R
pcDNA3
pGEM-7Zf(+)
pRACE (linearized)
pUAST
pcDNA3.1(+)
pGEM-7Zf(-)
pRANGER-BTB-1
pUC118
pcDNA3.1(-)
pGEM-9Zf(-)
pRANGER-BTB-2
pUC119
pCold I
pGEM-T
pRANGER-BTB-3
pUC18
pCold II
pGEM-T (linearized)
pRANGER-BTB-5
pUC19
pCold III
pGEM-T Easy
pRHA-109
pUC19L
pCold IV
pGEM-T Easy (linearized)
pRHA-113
pUC57
pCold TF
pGGA
pRHA-67
pUC57-Kan
pCR-XL-2-TOPO
pGGAselect
pRSET A
pUC57-Simple
pCR-XL-2-TOPO (linearized)
PGK promoter
pRSET B
pUCmu
pDD
pGS-21a
pRSET C
pVLT31
pDNR-Dual
pH6HTC His6HaloTag T7
pSB1C3
pVLT33
pDNR-LIB
pH6HTN His6HaloTag T7
pSC101
pVLT35
pE-SUMOpro Amp
pHP45(omega)
pSEAP2-Basic
pYES1L
pE-SUMOpro Kan
pHP45(omega)-Km
pSF-Core
pZErO-2
pE-SUMOpro3 Amp
pHP45(omega)-Tc
pSMART BAC v2.0
SNAP-tag
pE-SUMOpro3 Kan
pHSG298
pSMART BAC v2.0 (BamHI linearized)
SP6 promoter
pE-SUMOstar Amp
pHSG299
pSMART BAC v2.0 (EcoRI linearized)
SV40 promoter
pE-SUMOstar Kan
pHSG396
pSMART BAC v2.0 (HindIII linearized)
T3 promoter
pEASY-Blunt (linearized)
pHSG398
pSMART FOS v2.0 (linearized)
T5 promoter
pEASY-Blunt E1 (linearized)
pHY300PLK
pSMART HCAmp
T7 promoter
pEASY-Blunt E2 (linearized)
pICOz
pSMART HCAmp (linearized)
tac promoter
pEASY-Blunt Simple (linearized)
pInvRecA
pSMART HCKan
trc promoter
pEASY-Blunt Zero (linearized)
pJAZZ-OC
pSMART HCKan (linearized)
phiX174
pEASY-Blunt3 (linearized)
pJAZZ-OK

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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.
Try for Free