pETcoco-2

Bacterial vector with an ampicillin resistance marker that allows single-copy replication, on-demand amplification, and tightly regulated expression.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pETcoco-2
lac operator T7 promoter PciI (10,151) NcoI (9856) BsaI (9479) SpeI (9391) BseRI (8369) SnaBI (8300) PflFI - Tth111I (7940) KpnI (7880) Acc65I (7876) SbfI (6694) BsrGI (6449) FspAI (6421) RBS SphI (74) NsiI (76) NheI (96) BmtI (100) SrfI (181) AsiSI (188) ZraI (200) AatII (202) AscI (207) PmlI (215) BsiWI (219) BstBI (228) HindIII (231) NotI (238) PacI (288) Bsu36I (293) SwaI (302) AvrII (309) PspXI (320) 6xHis BamHI (1974) EcoRI (2050) SfiI (2434) PaqCI (2679) ScaI (3999) FseI (5567) SacII (5571) StuI (5843) pETcoco™-2 12,417 bp
PciI  (10,151)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
NcoI  (9856)
1 site		 C C A T G G G G T A C C
BsaI  (9479)
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.
SpeI  (9391)
1 site		 A C T A G T T G A T C A
BseRI  (8369)
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.
SnaBI  (8300)
1 site		 T A C G T A A T G C A T
PflFI  (7940)
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  (7940)
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.
KpnI  (7880)
1 site		 G G T A C C C C A T G G
Acc65I  (7876)
1 site		 G G T A C C C C A T G G
SbfI  (6694)
1 site		 C C T G C A G G G G A C G T C C
BsrGI  (6449)
1 site		 T G T A C A A C A T G T
BsrGI is typically used at 37°C, but is even more active at 60°C.
FspAI  (6421)
1 site		 R T G C G C A Y Y A C G C G T R
SphI  (74)
1 site		 G C A T G C C G T A C G
NsiI  (76)
1 site		 A T G C A T T A C G T A
NheI  (96)
1 site		 G C T A G C C G A T C G
BmtI  (100)
1 site		 G C T A G C C G A T C G
SrfI  (181)
1 site		 G C C C G G G C C G G G C C C G
AsiSI  (188)
1 site		 G C G A T C G C C G C T A G C G
ZraI  (200)
1 site		 G A C G T C C T G C A G
AatII  (202)
1 site		 G A C G T C C T G C A G
AscI  (207)
1 site		 G G C G C G C C C C G C G C G G
PmlI  (215)
1 site		 C A C G T G G T G C A C
BsiWI  (219)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
BstBI  (228)
1 site		 T T C G A A A A G C T T
HindIII  (231)
1 site		 A A G C T T T T C G A A
NotI  (238)
1 site		 G C G G C C G C C G C C G G C G
PacI  (288)
1 site		 T T A A T T A A A A T T A A T T
Bsu36I  (293)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
SwaI  (302)
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.
AvrII  (309)
1 site		 C C T A G G G G A T C C
PspXI  (320)
1 site		 V C T C G A G B B G A G C T C V
BamHI  (1974)
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  (2050)
1 site		 G A A T T C C T T A A G
SfiI  (2434)
1 site		 G G C C N N N N N G G C C C C G G N N N N N C C G G
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
PaqCI  (2679)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
ScaI  (3999)
1 site		 A G T A C T T C A T G A
FseI  (5567)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
SacII  (5571)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
StuI  (5843)
1 site		 A G G C C T T C C G G A
sopA
7400 .. 8575  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
sopA
7400 .. 8575  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
trfA
2080 .. 3228  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds to and activates oriV
trfA
2080 .. 3228  =  1149 bp
382 amino acids  =  43.8 kDa
Product: trans-acting replication protein that binds to and activates oriV
lacI
10,928 .. 12,010  =  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
10,928 .. 12,010  =  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).
araC
831 .. 1709  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
araC
831 .. 1709  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
AmpR
3446 .. 4306  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3446 .. 4234  =  789 bp
   262 amino acids  =  28.8 kDa
AmpR
3446 .. 4306  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4235 .. 4306  =  72 bp
   24 amino acids  =  2.8 kDa
AmpR
3446 .. 4306  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
repE
6066 .. 6821  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
repE
6066 .. 6821  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
oriV
5042 .. 5656  =  615 bp
origin of replication for the bacterial F plasmid
oriV
5042 .. 5656  =  615 bp
origin of replication for the bacterial F plasmid
sopC
9619 .. 10,092  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
sopC
9619 .. 10,092  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
cos
10,352 .. 10,750  =  399 bp
lambda cos site; allows packaging into phage lambda particles
cos
10,352 .. 10,750  =  399 bp
lambda cos site; allows packaging into phage lambda particles
incC
6824 .. 7074  =  251 bp
incompatibility region of the bacterial F plasmid
incC
6824 .. 7074  =  251 bp
incompatibility region of the bacterial F plasmid
MCS
70 .. 314  =  245 bp
multiple cloning site
MCS
70 .. 314  =  245 bp
multiple cloning site
ori2
5756 .. 5975  =  220 bp
secondary origin of replication for the bacterial F plasmid; also known as oriS
ori2
5756 .. 5975  =  220 bp
secondary origin of replication for the bacterial F plasmid; also known as oriS
araBAD promoter
1855 .. 2020  =  166 bp
promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction
araBAD promoter
1855 .. 2020  =  166 bp
promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction
AmpR promoter
4307 .. 4398  =  92 bp
AmpR promoter
4307 .. 4398  =  92 bp
lacI promoter
12,011 .. 12,088  =  78 bp
lacI promoter
12,011 .. 12,088  =  78 bp
T7 terminator
410 .. 457  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
410 .. 457  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
loxP
10,768 .. 10,801  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
10,768 .. 10,801  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
lac operator
3 .. 27  =  25 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
3 .. 27  =  25 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).
T7 promoter
12,401 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
12,401 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
326 .. 343  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
326 .. 343  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
58 .. 63  =  6 bp
ribosome binding site
RBS
58 .. 63  =  6 bp
ribosome binding site
sopB
8575 .. 9546  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F plasmid
sopB
8575 .. 9546  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F plasmid
ATG
72 .. 74  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
72 .. 74  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
75 .. 92  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
75 .. 92  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
S-Tag
102 .. 146  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
102 .. 146  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
enterokinase site
162 .. 176  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
162 .. 176  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
HSV tag
249 .. 281  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
249 .. 281  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
ORF:  2080 .. 3228  =  1149 bp
ORF:  382 amino acids  =  43.8 kDa
ORF:  4891 .. 5241  =  351 bp
ORF:  116 amino acids  =  12.8 kDa
ORF:  6055 .. 6285  =  231 bp
ORF:  76 amino acids  =  8.7 kDa
ORF:  6370 .. 6615  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  8575 .. 9546  =  972 bp
ORF:  323 amino acids  =  35.4 kDa
ORF:  10,738 .. 11,145  =  408 bp
ORF:  135 amino acids  =  14.9 kDa
ORF:  12,049 .. 12,312  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  2669 .. 2941  =  273 bp
ORF:  90 amino acids  =  10.3 kDa
ORF:  3200 .. 3427  =  228 bp
ORF:  75 amino acids  =  8.7 kDa
ORF:  7400 .. 8575  =  1176 bp
ORF:  391 amino acids  =  44.1 kDa
ORF:  8927 .. 9325  =  399 bp
ORF:  132 amino acids  =  15.6 kDa
ORF:  1395 .. 1643  =  249 bp
ORF:  82 amino acids  =  9.3 kDa
ORF:  2808 .. 3203  =  396 bp
ORF:  131 amino acids  =  14.1 kDa
ORF:  3576 .. 3842  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  6066 .. 6821  =  756 bp
ORF:  251 amino acids  =  29.4 kDa
ORF:  2821 .. 3216  =  396 bp
ORF:  131 amino acids  =  14.7 kDa
ORF:  11,760 .. 89  =  747 bp
ORF:  248 amino acids  =  26.1 kDa
ORF:  831 .. 1760  =  930 bp
ORF:  309 amino acids  =  35.2 kDa
ORF:  1941 .. 2609  =  669 bp
ORF:  222 amino acids  =  23.7 kDa
ORF:  6222 .. 6455  =  234 bp
ORF:  77 amino acids  =  9.1 kDa
ORF:  8616 .. 8852  =  237 bp
ORF:  78 amino acids  =  8.3 kDa
ORF:  10,908 .. 11,171  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3446 .. 4306  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  10,928 .. 11,887  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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Individual Sequences & Maps

pACYCDuet-1
pET-24a(+)
pET-3a
pET-51b(+) Ek_LIC (linearized)
pCDF-1b
pET-24b(+)
pET-3b
pET-52b(+)
pCDF-2 Ek_LIC
pET-24c(+)
pET-3c
pET-52b(+) 3C_LIC
pCDF-2 Ek_LIC (linearized)
pET-24d(+)
pET-3d
pET-52b(+) 3C_LIC (linearized)
pCDFDuet-1
pET-25b(+)
pET-40b(+)
pET-53-DEST
pCOLA-2-DEST
pET-26b(+)
pET-41 Ek_LIC
pET-54-DEST
pCOLA-3-DEST
pET-27b(+)
pET-41 Ek_LIC (linearized)
pET-55-DEST
pCOLADuet-1
pET-28a(+)
pET-41a(+)
pET-56-DEST
pET-11a
pET-28b(+)
pET-41b(+)
pET-57-DEST
pET-11b
pET-28c(+)
pET-41c(+)
pET-58-DEST
pET-11c
pET-29a(+)
pET-42a(+)
pET-59-DEST
pET-11d
pET-29b(+)
pET-42b(+)
pET-60-DEST
pET-14b
pET-29c(+)
pET-42c(+)
pET-61-DEST
pET-15b
pET-30 Ek_LIC
pET-43.1 Ek_LIC
pET-62-DEST
pET-16b
pET-30 Ek_LIC (linearized)
pET-43.1 Ek_LIC (linearized)
pET-9a
pET-17b
pET-30 Xa_LIC
pET-43.1a(+)
pET-9b
pET-19b
pET-30 Xa_LIC (linearized)
pET-43.1b(+)
pET-9c
pET-20b(+)
pET-30a(+)
pET-43.1c(+)
pET-9d
pET-21(+)
pET-30b(+)
pET-44a(+)
pETBlue-1
pET-21a(+)
pET-30c(+)
pET-44b(+)
pETBlue-2
pET-21b(+)
pET-31b(+)
pET-44c(+)
pETcoco-1
pET-21c(+)
pET-32 Ek_LIC
pET-45b(+)
pETcoco-2
pET-21d(+)
pET-32 Ek_LIC (linearized)
pET-46 Ek_LIC
pETDuet-1
pET-22b(+)
pET-32 Xa_LIC
pET-46 Ek_LIC (linearized)
pLacI
pET-23(+)
pET-32 Xa_LIC (linearized)
pET-47b(+)
pLysE
pET-23a(+)
pET-32a(+)
pET-48b(+)
pLysS
pET-23b(+)
pET-32b(+)
pET-49b(+)
pRSF-1b
pET-23c(+)
pET-32c(+)
pET-50b(+)
pRSF-2 Ek_LIC
pET-23d(+)
pET-33b(+)
pET-51b(+)
pRSF-2 Ek_LIC (linearized)
pET-24(+)
pET-39b(+)
pET-51b(+) Ek_LIC
pRSFDuet-1

Plasmid Sets

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I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
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Luciferase Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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