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Plasmid Files

LITMUS38

High copy-number E. coli vector with a multiple cloning site flanked by T7 promoters. For wild-type T7 promoters, use LITMUS38i.

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LITMUS38 Sequence and MapLITMUS38.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 PvuII (2701) AflII (2594) StuI (2592) AccI (2575) SalI (2574) SphI (2572) BsrGI (2561) BsmBI (2557) BspEI (2555) MluI (2548) EagI (2540) BmtI (2536) NheI (2532) EcoRI (2526) BamHI (2518) EcoRV (2513) PstI (2509) HindIII (2497) PluTI (2495) SfoI (2493) NarI (2492) KasI (2491) NaeI (2486) NgoMIV (2484) MfeI (2476) ApaI (2471) EcoO109I - PspOMI (2467) SpeI (2460) SnaBI (2447) lac operator PciI (2309) PspFI (2009) BseYI (2005) AlwNI (1900) HpaI (3) ScaI (449) AseI (755) NmeAIII (783) BpmI (861) BsaI (864) SwaI (1102) PsiI (1199) AvaI - BsoBI (1216) BmeT110I (1217) DraIII (1327) BtgZI (1328) LITMUS38 2814 bp
PvuII  (2701)
1 site
C A G C T G G T C G A C
AflII  (2594)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
StuI  (2592)
1 site
A G G C C T T C C G G A
AccI  (2575)
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.
SalI  (2574)
1 site
G T C G A C C A G C T G
SphI  (2572)
1 site
G C A T G C C G T A C G
BsrGI  (2561)
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.
BsmBI  (2557)
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.
BspEI  (2555)
1 site
T C C G G A A G G C C T
MluI  (2548)
1 site
A C G C G T T G C G C A
EagI  (2540)
1 site
C G G C C G G C C G G C
BmtI  (2536)
1 site
G C T A G C C G A T C G
NheI  (2532)
1 site
G C T A G C C G A T C G
EcoRI  (2526)
1 site
G A A T T C C T T A A G
BamHI  (2518)
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.
EcoRV  (2513)
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.
PstI  (2509)
1 site
C T G C A G G A C G T C
HindIII  (2497)
1 site
A A G C T T T T C G A A
PluTI  (2495)
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  (2493)
1 site
G G C G C C C C G C G G
NarI  (2492)
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.
KasI  (2491)
1 site
G G C G C C C C G C G G
NaeI  (2486)
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.
NgoMIV  (2484)
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.
MfeI  (2476)
1 site
C A A T T G G T T A A C
ApaI  (2471)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoO109I  (2467)
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.
PspOMI  (2467)
1 site
G G G C C C C C C G G G
SpeI  (2460)
1 site
A C T A G T T G A T C A
SnaBI  (2447)
1 site
T A C G T A A T G C A T
PciI  (2309)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2009)
1 site
C C C A G C G G G T C G
BseYI  (2005)
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.
AlwNI  (1900)
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.
HpaI  (3)
1 site
G T T A A C C A A T T G
ScaI  (449)
1 site
A G T A C T T C A T G A
AseI  (755)
1 site
A T T A A T T A A T T A
NmeAIII  (783)
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  (861)
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  (864)
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.
SwaI  (1102)
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.
PsiI  (1199)
1 site
T T A T A A A A T A T T
AvaI  (1216)
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  (1216)
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.
BmeT110I  (1217)
1 site
C Y C G R G G R G C Y C
DraIII  (1327)
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  (1328)
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.
AmpR
143 .. 1003  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   143 .. 211  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
143 .. 1003  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   212 .. 1003  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
143 .. 1003  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1665 .. 2253  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1665 .. 2253  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
M13 ori
1045 .. 1554  =  510 bp
M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
M13 ori
1045 .. 1554  =  510 bp
M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
lacZα
2424 .. 2786  =  363 bp
120 amino acids  =  13.7 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2424 .. 2786  =  363 bp
120 amino acids  =  13.7 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
38 .. 142  =  105 bp
AmpR promoter
38 .. 142  =  105 bp
lac promoter
2350 .. 2380  =  31 bp
   Segment 1:  -35  
   2350 .. 2355  =  6 bp
promoter for the E. coli lac operon
lac promoter
2350 .. 2380  =  31 bp
   Segment 2:  
   2356 .. 2373  =  18 bp
promoter for the E. coli lac operon
lac promoter
2350 .. 2380  =  31 bp
   Segment 3:  -10  
   2374 .. 2380  =  7 bp
promoter for the E. coli lac operon
lac promoter
2350 .. 2380  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
2388 .. 2404  =  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
2388 .. 2404  =  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).
MCS
2467 .. 2579  =  113 bp
multiple cloning site
MCS
2467 .. 2579  =  113 bp
multiple cloning site
T7 promoter
2449 .. 2466  =  18 bp
mutant version of the promoter for bacteriophage
T7 RNA polymerase
T7 promoter
2449 .. 2466  =  18 bp
mutant version of the promoter for bacteriophage
T7 RNA polymerase
T7 promoter
2594 .. 2611  =  18 bp
mutant version of the promoter for bacteriophage
T7 RNA polymerase
T7 promoter
2594 .. 2611  =  18 bp
mutant version of the promoter for bacteriophage
T7 RNA polymerase
M13 rev
2412 .. 2428  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2412 .. 2428  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
2615 .. 2631  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
2615 .. 2631  =  17 bp
common sequencing primer, one of multiple similar
variants
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