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Plasmid Files

Lafmid BA

Phagemid vector used for cDNA cloning.

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Lafmid BA Sequence and MapLafmid BA.dna
Map and Sequence File   
Sequence Author:  I.M.A.G.E. Consortium
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 BspDI - ClaI (0) XmnI (2947) BsaHI (2885) ScaI (2828) TatI (2826) NmeAIII (2496) BpmI (2418) BsaI (2409) AhdI (2348) BspHI (2175) AlwNI (1871) PspFI (1763) BseYI (1759) BciVI (1658) NdeI (36) SwaI (99) DraIII (324) BtgZI (325) BanII (399) NgoMIV (425) NaeI (427) EagI - NotI (1071) BamHI (1082) HindIII (1096) lac operator BspQI - SapI (1339) AflIII - PciI (1455) Lafmid BA 3152 bp
BspDI  (0)
1 site
A T C G A T T A G C T A
ClaI  (0)
1 site
A T C G A T T A G C T A
XmnI  (2947)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (2885)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
ScaI  (2828)
1 site
A G T A C T T C A T G A
TatI  (2826)
1 site
W G T A C W W C A T G W
NmeAIII  (2496)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BpmI  (2418)
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.
BsaI  (2409)
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.
AhdI  (2348)
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.
BspHI  (2175)
1 site
T C A T G A A G T A C T
AlwNI  (1871)
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.
PspFI  (1763)
1 site
C C C A G C G G G T C G
BseYI  (1759)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
BciVI  (1658)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
NdeI  (36)
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.
SwaI  (99)
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.
DraIII  (324)
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.
BtgZI  (325)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BanII  (399)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
NgoMIV  (425)
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  (427)
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.
EagI  (1071)
1 site
C G G C C G G C C G G C
NotI  (1071)
1 site
G C G G C C G C C G C C G G C G
BamHI  (1082)
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.
HindIII  (1096)
1 site
A A G C T T T T C G A A
BspQI  (1339)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1339)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
AflIII  (1455)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (1455)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2275 .. 3135  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2275 .. 3066  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2275 .. 3135  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3067 .. 3135  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2275 .. 3135  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1516 .. 2104  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1516 .. 2104  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
100 .. 555  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
100 .. 555  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
MCS
1070 .. 1101  =  32 bp
multiple cloning site
MCS
1070 .. 1101  =  32 bp
multiple cloning site
lac promoter
1162 .. 1192  =  31 bp
   Segment 3:  -10  
   1162 .. 1168  =  7 bp
promoter for the E. coli lac operon
lac promoter
1162 .. 1192  =  31 bp
   Segment 2:  
   1169 .. 1186  =  18 bp
promoter for the E. coli lac operon
lac promoter
1162 .. 1192  =  31 bp
   Segment 1:  -35  
   1187 .. 1192  =  6 bp
promoter for the E. coli lac operon
lac promoter
1162 .. 1192  =  31 bp
3 segments
promoter for the E. coli lac operon
M13 fwd
1042 .. 1058  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
1042 .. 1058  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
1114 .. 1130  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
1114 .. 1130  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
1138 .. 1154  =  17 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
1138 .. 1154  =  17 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).
ATG
1116 .. 1118  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1116 .. 1118  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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