Resources
Plasmid Files

pHM3C-LIC

Bacterial vector for expressing a protein with an N-terminal 6xHis-MBP cassette followed by an HRV 3C protease cleavage site.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pHM3C-LIC Sequence and MappHM3C-LIC.dna
Map and Sequence File   
Sequence Author:  Center for Structures of Membrane Proteins
Download Free Trial Get SnapGene Viewer

 SalI (6344) EagI - NotI (6334) SmaI (6310) TspMI - XmaI (6308) EcoRI (6254) SacI (6221) Eco53kI (6219) BmgBI (5653) BsiWI (5401) BsaI (5174) NcoI (5098) ATG NdeI (5070) XbaI (5030) T7 promoter SgrAI (4923) SphI (4775) MluI (4242) BstEII (4060) NmeAIII (4042) ApaI (4039) PspOMI (4035) EcoRV (3796) PshAI (3401) BglI (3185) PaeR7I - PspXI - XhoI (6371) DraIII (245) AsiSI (945) NruI (1286) BssS α I (1968) PciI (2141) BspQI - SapI (2258) TatI (2337) BstZ17I (2374) PflFI - Tth111I (2399) PpuMI (3136) FspI - FspAI (3164) pHM3C-LIC 6533 bp
SalI  (6344)
1 site
G T C G A C C A G C T G
EagI  (6334)
1 site
C G G C C G G C C G G C
NotI  (6334)
1 site
G C G G C C G C C G C C G G C G
SmaI  (6310)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (6308)
1 site
C C C G G G G G G C C C
XmaI  (6308)
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.
EcoRI  (6254)
1 site
G A A T T C C T T A A G
SacI  (6221)
1 site
G A G C T C C T C G A G
Eco53kI  (6219)
1 site
G A G C T C C T C G A G
BmgBI  (5653)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
BsiWI  (5401)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BsaI  (5174)
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.
NcoI  (5098)
1 site
C C A T G G G G T A C C
NdeI  (5070)
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.
XbaI  (5030)
1 site
T C T A G A A G A T C T
SgrAI  (4923)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
SphI  (4775)
1 site
G C A T G C C G T A C G
MluI  (4242)
1 site
A C G C G T T G C G C A
BstEII  (4060)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
NmeAIII  (4042)
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).
ApaI  (4039)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4035)
1 site
G G G C C C C C C G G G
EcoRV  (3796)
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.
PshAI  (3401)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BglI  (3185)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
PaeR7I  (6371)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (6371)
1 site
V C T C G A G B B G A G C T C V
XhoI  (6371)
1 site
C T C G A G G A G C T C
DraIII  (245)
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.
AsiSI  (945)
1 site
G C G A T C G C C G C T A G C G
NruI  (1286)
1 site
T C G C G A A G C G C T
BssSαI  (1968)
1 site
C A C G A G G T G C T C
PciI  (2141)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2258)
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  (2258)
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.
TatI  (2337)
1 site
W G T A C W W C A T G W
BstZ17I  (2374)
1 site
G T A T A C C A T A T G
PflFI  (2399)
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  (2399)
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.
PpuMI  (3136)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
FspI  (3164)
1 site
T G C G C A A C G C G T
FspAI  (3164)
1 site
R T G C G C A Y Y A C G C G T R
MBP
5111 .. 6211  =  1101 bp
367 amino acids  =  40.3 kDa
Product: maltose binding protein from E. coli
This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol.
MBP
5111 .. 6211  =  1101 bp
367 amino acids  =  40.3 kDa
Product: maltose binding protein from E. coli
This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol.
lacI
3515 .. 4597  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
3515 .. 4597  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
KanR
560 .. 1375  =  816 bp
271 amino acids  =  31.1 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
560 .. 1375  =  816 bp
271 amino acids  =  31.1 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
ori
1497 .. 2085  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1497 .. 2085  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
rop
2515 .. 2706  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
2515 .. 2706  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
lacI promoter
4598 .. 4675  =  78 bp
lacI promoter
4598 .. 4675  =  78 bp
T7 terminator
6461 .. 6508  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
6461 .. 6508  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
5003 .. 5027  =  25 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
5003 .. 5027  =  25 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).
HRV 3C site
6278 .. 6301  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human
rhinovirus 3C and PreScission proteases
HRV 3C site
6278 .. 6301  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human
rhinovirus 3C and PreScission proteases
T7 promoter
4984 .. 5002  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4984 .. 5002  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
5084 .. 5101  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5084 .. 5101  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
thrombin site
6314 .. 6331  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
6314 .. 6331  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
6xHis
6377 .. 6394  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
6377 .. 6394  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
5058 .. 5063  =  6 bp
ribosome binding site
RBS
5058 .. 5063  =  6 bp
ribosome binding site
ATG
5072 .. 5074  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5072 .. 5074  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter