pALTER-MAX

Mammalian expression vector with the CMV enhancer/promoter and with ampicillin and chloramphenicol resistance markers.

Sequence Author: Promega

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BglII (5533) AlwNI (5112) AhdI (4633) XmnI (4033) EcoO109I (3653) PfoI (3594) BspEI (2832) BsrGI (96) SpeI (152) NdeI (387) SnaBI (493) AsiSI (664) Eco53kI (727) SacI (729) I-PpoI (851) BfuAI - BspMI (877) BbsI (961) PaeR7I - XhoI (1091) EcoRI (1096) AflIII - MluI (1102) Acc65I (1108) KpnI (1112) XbaI (1114) SalI (1120) AccI (1121) TspMI - XmaI (1125) SmaI (1127) EagI - NotI (1131) T3 promoter HpaI (1301) MfeI (1310) NgoMIV (1612) NaeI (1614) DraIII (1720) CsiI - SexAI * (2123) SfiI (2309) BseRI (2352) StuI (2355) AvrII (2356) Bpu10I (2609) pALTER®-MAX 5538 bp
BglII  (5533)
1 site
A G A T C T T C T A G A
AlwNI  (5112)
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  (4633)
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.
XmnI  (4033)
1 site
G A A N N N N T T C C T T N N N N A A G
EcoO109I  (3653)
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  (3594)
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.
BspEI  (2832)
1 site
T C C G G A A G G C C T
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.
BfuAI  (877)
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  (877)
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.
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.
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
Acc65I  (1108)
1 site
G G T A C C C C A T G G
KpnI  (1112)
1 site
G G T A C C C C A T G G
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.
EagI  (1131)
1 site
C G G C C G G C C G G C
NotI  (1131)
1 site
G C G G C C G C C G C C G G C G
HpaI  (1301)
1 site
G T T A A C C A A T T G
MfeI  (1310)
1 site
C A A T T G G T T A A C
NgoMIV  (1612)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (1614)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
DraIII  (1720)
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  (2123)
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  (2123)
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  (2309)
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  (2352)
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  (2355)
1 site
A G G C C T T C C G G A
AvrII  (2356)
1 site
C C T A G G G G A T C C
Bpu10I  (2609)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
AmpR
3846 .. 4706  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   3846 .. 3914  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3846 .. 4706  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   3915 .. 4706  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3846 .. 4706  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
2623 .. 3282  =  660 bp
219 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2623 .. 3282  =  660 bp
219 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
4877 .. 5465  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4877 .. 5465  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1487 .. 1942  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1487 .. 1942  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
2014 .. 2371  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2014 .. 2371  =  358 bp
SV40 enhancer and early promoter
CMV promoter
518 .. 729  =  212 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
518 .. 729  =  212 bp
human cytomegalovirus (CMV) immediate early promoter
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
1180 .. 1301  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1180 .. 1301  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3741 .. 3845  =  105 bp
AmpR promoter
3741 .. 3845  =  105 bp
cat promoter
2520 .. 2622  =  103 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
cat promoter
2520 .. 2622  =  103 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
poly(A) signal
3379 .. 3427  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3379 .. 3427  =  49 bp
synthetic polyadenylation signal
MCS
1091 .. 1137  =  47 bp
multiple cloning site
MCS
1091 .. 1137  =  47 bp
multiple cloning site
T7 promoter
1067 .. 1085  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1067 .. 1085  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
1145 .. 1162  =  18 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
1145 .. 1162  =  18 bp
promoter for bacteriophage T3 RNA polymerase
SV40 ori
2222 .. 2357  =  136 bp
SV40 origin of replication
SV40 ori
2222 .. 2357  =  136 bp
SV40 origin of replication
ORF:  2623 .. 3282  =  660 bp
ORF:  219 amino acids  =  25.6 kDa
ORF:  3846 .. 4706  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2287 .. 2553  =  267 bp
ORF:  88 amino acids  =  9.9 kDa
ORF:  4310 .. 4576  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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