pCDFDuet-1

Bacterial vector with a CloDF13 (CDF) origin for the co-expression of two genes.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pCDFDuet-1
NcoI (69) RBS lac operator T7 promoter EcoNI (3754) ACYCDuetUP1 Primer (3722 .. 3740) PfoI (3718) AclI (3626) BstAPI (3610) MluI (3286) ApaI (3083) PspOMI (3079) HpaI (2784) BsmBI - Esp3I (2671) PluTI (2651) SfoI (2649) NarI * (2648) KasI (2647) XbaI (2366) BamHI (106) EcoRI (112) Eco53kI (120) SacI (122) AscI (125) PstI - SbfI (135) SalI (137) HindIII (143) NotI (150) AflII (163) DuetUP2 Primer (189 .. 208) BsrGI - TatI (190) DuetDOWN1 Primer (189 .. 208) T7 promoter lac operator RBS NdeI (298) BglII (305) MfeI (311) EcoRV (319) FseI (328) AsiSI (337) ZraI (344) AatII (346) Acc65I (348) KpnI (352) AvaI - BsoBI - PaeR7I - PspXI - XhoI (354) PacI (429) AvrII (433) T7 Terminator Primer (448 .. 466) EcoO109I (478) T7 terminator Bsu36I (517) AgeI (566) DrdI - PflFI - Tth111I (626) BsaAI (715) BglI (843) SspI (1488) BspHI (1519) NheI (1566) BmtI (1570) PciI (1666) NspI (1670) SgrAI (1693) pCDFDuet™-1 3781 bp
NcoI  (69)
1 site		 C C A T G G G G T A C C
EcoNI  (3754)
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.
PfoI  (3718)
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.
AclI  (3626)
1 site		 A A C G T T T T G C A A
BstAPI  (3610)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
MluI  (3286)
1 site		 A C G C G T T G C G C A
ApaI  (3083)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (3079)
1 site		 G G G C C C C C C G G G
HpaI  (2784)
1 site		 G T T A A C C A A T T G
BsmBI  (2671)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2671)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
PluTI  (2651)
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  (2649)
1 site		 G G C G C C C C G C G G
NarI  (2648)
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  (2647)
1 site		 G G C G C C C C G C G G
XbaI  (2366)
1 site		 T C T A G A A G A T C T
BamHI  (106)
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  (112)
1 site		 G A A T T C C T T A A G
Eco53kI  (120)
1 site		 G A G C T C C T C G A G
SacI  (122)
1 site		 G A G C T C C T C G A G
AscI  (125)
1 site		 G G C G C G C C C C G C G C G G
PstI  (135)
1 site		 C T G C A G G A C G T C
SbfI  (135)
1 site		 C C T G C A G G G G A C G T C C
SalI  (137)
1 site		 G T C G A C C A G C T G
HindIII  (143)
1 site		 A A G C T T T T C G A A
NotI  (150)
1 site		 G C G G C C G C C G C C G G C G
AflII  (163)
1 site		 C T T A A G G A A T T C
BsrGI  (190)
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.
TatI  (190)
1 site		 W G T A C W W C A T G W
NdeI  (298)
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.
BglII  (305)
1 site		 A G A T C T T C T A G A
MfeI  (311)
1 site		 C A A T T G G T T A A C
EcoRV  (319)
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.
FseI  (328)
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.
AsiSI  (337)
1 site		 G C G A T C G C C G C T A G C G
ZraI  (344)
1 site		 G A C G T C C T G C A G
AatII  (346)
1 site		 G A C G T C C T G C A G
Acc65I  (348)
1 site		 G G T A C C C C A T G G
KpnI  (352)
1 site		 G G T A C C C C A T G G
AvaI  (354)
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  (354)
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  (354)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (354)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (354)
1 site		 C T C G A G G A G C T C
PacI  (429)
1 site		 T T A A T T A A A A T T A A T T
AvrII  (433)
1 site		 C C T A G G G G A T C C
EcoO109I  (478)
1 site		 R G G N C C Y Y C C N G G R
Sticky ends from different EcoO109I sites may not be compatible.
Bsu36I  (517)
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.
AgeI  (566)
1 site		 A C C G G T T G G C C A
DrdI  (626)
1 site		 G A C N N N N N N G T C C T G N N N N N N C A G
Sticky ends from different DrdI sites may not be compatible.
PflFI  (626)
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  (626)
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  (715)
1 site		 Y A C G T R R T G C A Y
BglI  (843)
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.
SspI  (1488)
1 site		 A A T A T T T T A T A A
BspHI  (1519)
1 site		 T C A T G A A G T A C T
NheI  (1566)
1 site		 G C T A G C C G A T C G
BmtI  (1570)
1 site		 G C T A G C C G A T C G
PciI  (1666)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
NspI  (1670)
1 site		 R C A T G Y Y G T A C R
SgrAI  (1693)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
ACYCDuetUP1 Primer
19-mer  /  63% GC
1 binding site
3722 .. 3740  =  19 annealed bases
Tm  =  60°C
DuetUP2 Primer
20-mer  /  50% GC
1 binding site
189 .. 208  =  20 annealed bases
Tm  =  57°C
DuetDOWN1 Primer
20-mer  /  50% GC
1 binding site
189 .. 208  =  20 annealed bases
Tm  =  57°C
T7 Terminator Primer
19-mer  /  53% GC
1 binding site
448 .. 466  =  19 annealed bases
Tm  =  57°C
lacI
2559 .. 3641  =  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
2559 .. 3641  =  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).
SmR
680 .. 1471  =  792 bp
263 amino acids  =  29.3 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and streptomycin
SmR
680 .. 1471  =  792 bp
263 amino acids  =  29.3 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and streptomycin
CloDF13 ori
1611 .. 2349  =  739 bp
Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins.
CloDF13 ori
1611 .. 2349  =  739 bp
Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins.
MCS-2
297 .. 438  =  142 bp
multiple cloning site 2
MCS-2
297 .. 438  =  142 bp
multiple cloning site 2
MCS-1
69 .. 168  =  100 bp
multiple cloning site 1
MCS-1
69 .. 168  =  100 bp
multiple cloning site 1
AmpR promoter
1472 .. 1563  =  92 bp
AmpR promoter
1472 .. 1563  =  92 bp
lacI promoter
3642 .. 3719  =  78 bp
lacI promoter
3642 .. 3719  =  78 bp
T7 terminator
462 .. 509  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
462 .. 509  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
233 .. 257  =  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
233 .. 257  =  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).
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
3765 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3765 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
214 .. 232  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
214 .. 232  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
58 .. 63  =  6 bp
ribosome binding site
RBS
58 .. 63  =  6 bp
ribosome binding site
RBS
286 .. 291  =  6 bp
ribosome binding site
RBS
286 .. 291  =  6 bp
ribosome binding site
ATG
300 .. 302  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
300 .. 302  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
S-Tag
366 .. 410  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
366 .. 410  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
ATG
71 .. 73  =  3 bp
1 amino acid  =  149.2 Da
ATG
71 .. 73  =  3 bp
1 amino acid  =  149.2 Da
6xHis
83 .. 100  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
83 .. 100  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ORF:  1049 .. 1432  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  2525 .. 2776  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  894 .. 1172  =  279 bp
ORF:  92 amino acids  =  9.4 kDa
ORF:  1182 .. 1439  =  258 bp
ORF:  85 amino acids  =  9.1 kDa
ORF:  680 .. 1471  =  792 bp
ORF:  263 amino acids  =  29.3 kDa
ORF:  2539 .. 2802  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3391 .. 158  =  549 bp
ORF:  182 amino acids  =  19.6 kDa
ORF:  2559 .. 3518  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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Download pCDFDuet-1.dna file

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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

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