pCI-neo

Mammalian cell expression vector with the CMV promoter and a neomycin (G418) resistance marker.

Sequence Author: Promega

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HomePlasmidsMammalian Expression VectorspCI-neo
BglII (5467) AlwNI (5046) AhdI (4567) BpmI (4498) XmnI (3967) EcoO109I (3587) PfoI (3528) BamHI (3385) BstXI (3312) BstBI (3273) RsrII (3107) BssHII (2988) PflFI - Tth111I (2709) BsrGI (96) SpeI (152) NdeI (387) SnaBI (493) AsiSI (664) minimal CMV promoter Eco53kI (727) SacI (729) I-PpoI (851) BbsI (961) NheI (1085) BmtI (1089) PaeR7I - XhoI (1091) EcoRI (1096) AflIII - MluI (1102) XbaI (1114) SalI (1120) AccI (1121) TspMI - XmaI (1125) SmaI (1127) NotI (1131) T3 promoter HpaI (1297) MfeI (1306) DraIII (1716) CsiI - SexAI * (2164) SfiI (2350) BseRI (2393) StuI (2396) AvrII (2397) KasI (2590) NarI (2591) SfoI (2592) PluTI (2594) pCI-neo 5472 bp
BglII  (5467)
1 site		 A G A T C T T C T A G A
AlwNI  (5046)
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.
AhdI  (4567)
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.
BpmI  (4498)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
XmnI  (3967)
1 site		 G A A N N N N T T C C T T N N N N A A G
EcoO109I  (3587)
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.
PfoI  (3528)
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.
BamHI  (3385)
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.
BstXI  (3312)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
BstBI  (3273)
1 site		 T T C G A A A A G C T T
RsrII  (3107)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BssHII  (2988)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
PflFI  (2709)
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  (2709)
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.
BsrGI  (96)
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.
SpeI  (152)
1 site		 A C T A G T T G A T C A
NdeI  (387)
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.
SnaBI  (493)
1 site		 T A C G T A A T G C A T
AsiSI  (664)
1 site		 G C G A T C G C C G C T A G C G
Eco53kI  (727)
1 site		 G A G C T C C T C G A G
SacI  (729)
1 site		 G A G C T C C T C G A G
I-PpoI  (851)
1 site		 C T C T C T T A A G G T A G C G A G A G A A T T C C A T C G
I-PpoI is a homing endonuclease that can recognize a variety of similar recognition sequences.
BbsI  (961)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
NheI  (1085)
1 site		 G C T A G C C G A T C G
BmtI  (1089)
1 site		 G C T A G C C G A T C G
PaeR7I  (1091)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1091)
1 site		 C T C G A G G A G C T C
EcoRI  (1096)
1 site		 G A A T T C C T T A A G
AflIII  (1102)
1 site		 A C R Y G T T G Y R C A
Sticky ends from different AflIII sites may not be compatible.
MluI  (1102)
1 site		 A C G C G T T G C G C A
XbaI  (1114)
1 site		 T C T A G A A G A T C T
SalI  (1120)
1 site		 G T C G A C C A G C T G
AccI  (1121)
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.
TspMI  (1125)
1 site		 C C C G G G G G G C C C
XmaI  (1125)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (1127)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
NotI  (1131)
1 site		 G C G G C C G C C G C C G G C G
HpaI  (1297)
1 site		 G T T A A C C A A T T G
MfeI  (1306)
1 site		 C A A T T G G T T A A C
DraIII  (1716)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
CsiI  (2164)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (2164)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (2350)
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.
BseRI  (2393)
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.
StuI  (2396)
1 site		 A G G C C T T C C G G A
AvrII  (2397)
1 site		 C C T A G G G G A T C C
KasI  (2590)
1 site		 G G C G C C C C G C G G
NarI  (2591)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2592)
1 site		 G G C G C C C C G C G G
PluTI  (2594)
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.
AmpR
3780 .. 4640  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3780 .. 3848  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3780 .. 4640  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3849 .. 4640  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3780 .. 4640  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2463 .. 3257  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2463 .. 3257  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
4811 .. 5399  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4811 .. 5399  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1483 .. 1938  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1483 .. 1938  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2055 .. 2412  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2055 .. 2412  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
213 .. 517  =  305 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
213 .. 517  =  305 bp
human cytomegalovirus immediate early enhancer
chimeric intron
890 .. 1022  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
chimeric intron
890 .. 1022  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
SV40 poly(A) signal
1176 .. 1297  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1176 .. 1297  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3675 .. 3779  =  105 bp
AmpR promoter
3675 .. 3779  =  105 bp
MCS
1085 .. 1137  =  53 bp
multiple cloning site
MCS
1085 .. 1137  =  53 bp
multiple cloning site
poly(A) signal
3321 .. 3369  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3321 .. 3369  =  49 bp
synthetic polyadenylation signal
minimal CMV promoter
691 .. 729  =  39 bp
human cytomegalovirus (CMV) immediate early promoter
minimal CMV promoter
691 .. 729  =  39 bp
human cytomegalovirus (CMV) immediate early promoter
T3 promoter
1141 .. 1158  =  18 bp
promoter for bacteriophage T3 RNA polymerase (shorter by one base than the standard T3 promoter)
T3 promoter
1141 .. 1158  =  18 bp
promoter for bacteriophage T3 RNA polymerase (shorter by one base than the standard T3 promoter)
SV40 ori
2263 .. 2398  =  136 bp
SV40 origin of replication
SV40 ori
2263 .. 2398  =  136 bp
SV40 origin of replication
T7 promoter
1067 .. 1085  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1067 .. 1085  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  2635 .. 3021  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2463 .. 3257  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3780 .. 4640  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2772 .. 3308  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  4244 .. 4510  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Individual Sequences & Maps

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p3xFLAG-CMV-7
pcDNA6 TR
pCMV6-AN-DDK
pHom-1
p3xFLAG-CMV-7.1
pcDNA6 V5-His A
pCMV6-AN-FC
pHom-Mem1
p3xFLAG-CMV-8
pcDNA6 V5-His B
pCMV6-AN-FC-S
pHTC HaloTag CMV-neo
p3xFLAG-CMV-9
pcDNA6 V5-His C