pMCSG73

Compact bacterial vector encoding tRNA genes for rare Arg and Ile codons, with a NusA-TVMV-6xHis-Strep-Tag® II-TEV leader.

Sequence Author: Midwest Center for Structural Genomics

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BspEI (2) ScaI (5532) FspI (5274) BglI (5172) AhdI (5052) AlwNI (4575) DrdI (4267) PciI (4159) BspQI - SapI (4043) BstZ17I (3930) BsaAI (3911) PshAI (3904) BsmBI - Esp3I (3674) HpaI (3565) ApaI (3270) PspOMI (3266) BstEII (3240) SspI (151) TEV site Strep-Tag II 6xHis TVMV site BglII (333) BsmI (377) MscI (465) StuI (518) ZraI (1576) AatII (1578) NdeI (1707) XbaI (1744) T7 promoter AgeI (1855) AscI (2072) SbfI (2083) MreI (2220) AfeI (2306) PsiI (2505) EcoNI (2594) PfoI (2626) PflMI (2641) MluI (3059) pMCSG73 6227 bp
BspEI  (2)
1 site
T C C G G A A G G C C T
ScaI  (5532)
1 site
A G T A C T T C A T G A
FspI  (5274)
1 site
T G C G C A A C G C G T
BglI  (5172)
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.
AhdI  (5052)
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.
AlwNI  (4575)
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.
DrdI  (4267)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PciI  (4159)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4043)
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  (4043)
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.
BstZ17I  (3930)
1 site
G T A T A C C A T A T G
BsaAI  (3911)
1 site
Y A C G T R R T G C A Y
PshAI  (3904)
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.
BsmBI  (3674)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (3674)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
HpaI  (3565)
1 site
G T T A A C C A A T T G
ApaI  (3270)
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  (3266)
1 site
G G G C C C C C C G G G
BstEII  (3240)
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.
SspI  (151)
1 site
A A T A T T T T A T A A
BglII  (333)
1 site
A G A T C T T C T A G A
BsmI  (377)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
MscI  (465)
1 site
T G G C C A A C C G G T
StuI  (518)
1 site
A G G C C T T C C G G A
ZraI  (1576)
1 site
G A C G T C C T G C A G
AatII  (1578)
1 site
G A C G T C C T G C A G
NdeI  (1707)
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  (1744)
1 site
T C T A G A A G A T C T
AgeI  (1855)
1 site
A C C G G T T G G C C A
AscI  (2072)
1 site
G G C G C G C C C C G C G C G G
SbfI  (2083)
1 site
C C T G C A G G G G A C G T C C
MreI  (2220)
1 site
C G C C G G C G G C G G C C G C
AfeI  (2306)
1 site
A G C G C T T C G C G A
PsiI  (2505)
1 site
T T A T A A A A T A T T
EcoNI  (2594)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PfoI  (2626)
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.
PflMI  (2641)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
MluI  (3059)
1 site
A C G C G T T G C G C A
TEV site
155 .. 175  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
155 .. 175  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
Strep-Tag II
176 .. 199  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
Strep-Tag II
176 .. 199  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
6xHis
200 .. 217  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
200 .. 217  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
TVMV site
218 .. 238  =  21 bp
7 amino acids  =  865.9 Da
Product:
tobacco vein mottling virus (TVMV) NIa protease recognition and cleavage site
TVMV site
218 .. 238  =  21 bp
7 amino acids  =  865.9 Da
Product:
tobacco vein mottling virus (TVMV) NIa protease recognition and cleavage site
NusA
248 .. 1708  =  1461 bp
487 amino acids  =  53.9 kDa
Product: transcription elongation protein (N utilization substance protein A)
highly soluble in E. coli
NusA
248 .. 1708  =  1461 bp
487 amino acids  =  53.9 kDa
Product: transcription elongation protein (N utilization substance protein A)
highly soluble in E. coli
lacI
2709 .. 3791  =  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
2709 .. 3791  =  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).
AmpR
4979 .. 5839  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   4979 .. 5770  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4979 .. 5839  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   5771 .. 5839  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4979 .. 5839  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
4220 .. 4808  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4220 .. 4808  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
5840 .. 5943  =  104 bp
AmpR promoter
5840 .. 5943  =  104 bp
lacI promoter
2631 .. 2708  =  78 bp
lacI promoter
2631 .. 2708  =  78 bp
argU
2070 .. 2146  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
argU
2070 .. 2146  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
ileX
2416 .. 2491  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
ileX
2416 .. 2491  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
1752 .. 1776  =  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
1752 .. 1776  =  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).
T7 promoter
1777 .. 1795  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1777 .. 1795  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
1717 .. 1722  =  6 bp
ribosome binding site
RBS
1717 .. 1722  =  6 bp
ribosome binding site
ORF:  1630 .. 1920  =  291 bp
ORF:  96 amino acids  =  10.9 kDa
ORF:  134 .. 1522  =  1389 bp
ORF:  462 amino acids  =  53.6 kDa
ORF:  2492 .. 2959  =  468 bp
ORF:  155 amino acids  =  16.8 kDa
ORF:  3548 .. 3811  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  318 .. 554  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2205 .. 2459  =  255 bp
ORF:  84 amino acids  =  8.5 kDa
ORF:  2832 .. 3791  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  5109 .. 5375  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  137 .. 1708  =  1572 bp
ORF:  523 amino acids  =  58.3 kDa
ORF:  4979 .. 5839  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  3574 .. 3825  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
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Download pMCSG73.dna file

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