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Plasmid Files

pALTER®-MAX

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

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pALTER-MAX Sequence and MappALTER-MAX.dna
Map and Sequence File   
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) T7 promoter PaeR7I - XhoI (1091) EcoRI (1096) AflIII - MluI (1102) Acc65I (1108) KpnI (1112) MCS XbaI (1114) SalI (1120) AccI (1121) TspMI - XmaI (1125) SmaI (1127) EagI - NotI (1131) T3 promoter SV40 poly(A) signal HpaI (1301) MfeI (1310) NgoMIV (1612) NaeI (1614) DraIII (1720) 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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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.
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

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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
   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
   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
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