pMCSG12 (linearized)

Linearized bacterial coexpression vector for ligation-independent cloning (LIC), with a 6xHis-S-loop-TEV leader and a p15A origin. 

Sequence Author: Midwest Center for Structural Genomics

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HomePlasmidsStructural Genomics VectorspMCSG12 (linearized)
T7 promoter EcoNI (3684) BstAPI (3540) ApaLI (3236) AflIII - MluI (3216) BclI * (3202) BstEII (3034) ApaI (3013) PspOMI (3009) BciVI (2764) HpaI (2714) PluTI (2581) SfoI (2579) NarI * (2578) KasI (2577) AcuI (2356) XbaI (2296) NspI (2033) BssSI - BssSαI (1831) SacII (1707) SgrAI (1541) RBS NcoI (3780) ATG BfuAI - BspMI (3835) AscI (3836) PstI - SbfI (3846) AflII (3874) BsrGI (3901) T7 promoter lac operator RBS NdeI (4009) 6xHis BglII (4043) Acc65I (4104) KpnI (4108) End (4118) TEV site Start (0) AvaI - BsoBI - PaeR7I - PspXI - XhoI (57) PacI (132) AvrII (136) BlpI (154) EcoO109I (181) Bsu36I (220) DrdI - PflFI - Tth111I (329) ScaI (460) MscI (610) BspEI (877) Bpu10I (1101) BsaAI (1184) AfeI (1454) NheI (1455) BmtI (1459) BstZ17I (1468) XmnI (1511) pMCSG12 4118 bp
EcoNI  (3684)
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.
BstAPI  (3540)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
ApaLI  (3236)
1 site		 G T G C A C C A C G T G
AflIII  (3216)
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  (3216)
1 site		 A C G C G T T G C G C A
BclI  (3202)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (3034)
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.
ApaI  (3013)
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  (3009)
1 site		 G G G C C C C C C G G G
BciVI  (2764)
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.
HpaI  (2714)
1 site		 G T T A A C C A A T T G
PluTI  (2581)
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  (2579)
1 site		 G G C G C C C C G C G G
NarI  (2578)
1 site		 G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (2577)
1 site		 G G C G C C C C G C G G
AcuI  (2356)
1 site		 C T G A A G ( N ) 14 N N G A C T T C ( N ) 14
Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
XbaI  (2296)
1 site		 T C T A G A A G A T C T
NspI  (2033)
1 site		 R C A T G Y Y G T A C R
BssSI  (1831)
1 site		 C A C G A G G T G C T C
BssSαI  (1831)
1 site		 C A C G A G G T G C T C
SacII  (1707)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
SgrAI  (1541)
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.
NcoI  (3780)
1 site		 C C A T G G G G T A C C
BfuAI  (3835)
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  (3835)
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.
AscI  (3836)
1 site		 G G C G C G C C C C G C G C G G
PstI  (3846)
1 site		 C T G C A G G A C G T C
SbfI  (3846)
1 site		 C C T G C A G G G G A C G T C C
AflII  (3874)
1 site		 C T T A A G G A A T T C
BsrGI  (3901)
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.
NdeI  (4009)
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.
BglII  (4043)
1 site		 A G A T C T T C T A G A
Acc65I  (4104)
1 site		 G G T A C C C C A T G G
KpnI  (4108)
1 site		 G G T A C C C C A T G G
End  (4118)
0 sites
Start  (0)
0 sites
AvaI  (57)
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  (57)
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.
PaeR7I  (57)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (57)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (57)
1 site		 C T C G A G G A G C T C
PacI  (132)
1 site		 T T A A T T A A A A T T A A T T
AvrII  (136)
1 site		 C C T A G G G G A T C C
BlpI  (154)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
EcoO109I  (181)
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.
Bsu36I  (220)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
DrdI  (329)
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.
PflFI  (329)
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  (329)
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.
ScaI  (460)
1 site		 A G T A C T T C A T G A
MscI  (610)
1 site		 T G G C C A A C C G G T
BspEI  (877)
1 site		 T C C G G A A G G C C T
Bpu10I  (1101)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
BsaAI  (1184)
1 site		 Y A C G T R R T G C A Y
AfeI  (1454)
1 site		 A G C G C T T C G C G A
NheI  (1455)
1 site		 G C T A G C C G A T C G
BmtI  (1459)
1 site		 G C T A G C C G A T C G
BstZ17I  (1468)
1 site		 G T A T A C C A T A T G
XmnI  (1511)
1 site		 G A A N N N N T T C C T T N N N N A A G
lacI
2489 .. 3571  =  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
2489 .. 3571  =  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).
CmR
432 .. 1091  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
432 .. 1091  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
p15A ori
1720 .. 2264  =  545 bp
Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin.
p15A ori
1720 .. 2264  =  545 bp
Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin.
ATG
4011 .. 4013  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4011 .. 4013  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
4014 .. 4031  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4014 .. 4031  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
S loop
4050 .. 4103  =  54 bp
18 amino acids  =  1.7 kDa
Product: GroES chaperone mobile loop that interacts with GroEL
S loop
4050 .. 4103  =  54 bp
18 amino acids  =  1.7 kDa
Product: GroES chaperone mobile loop that interacts with GroEL
TEV site
4110 .. 4130  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
4110 .. 4130  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
cat promoter
1092 .. 1194  =  103 bp
promoter of the E. coli cat gene
cat promoter
1092 .. 1194  =  103 bp
promoter of the E. coli cat gene
lacI promoter
3572 .. 3649  =  78 bp
lacI promoter
3572 .. 3649  =  78 bp
T7 terminator
165 .. 212  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
165 .. 212  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
S-Tag
69 .. 113  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
69 .. 113  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
ATG
3782 .. 3784  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3782 .. 3784  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
3794 .. 3811  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3794 .. 3811  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac operator
3944 .. 3968  =  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
3944 .. 3968  =  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
3714 .. 3738  =  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
3714 .. 3738  =  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
3925 .. 3943  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3925 .. 3943  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3695 .. 3713  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3695 .. 3713  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
3769 .. 3774  =  6 bp
ribosome binding site
RBS
3769 .. 3774  =  6 bp
ribosome binding site
RBS
3997 .. 4002  =  6 bp
ribosome binding site
RBS
3997 .. 4002  =  6 bp
ribosome binding site
ORF:  2455 .. 2706  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  1635 .. 1913  =  279 bp
ORF:  92 amino acids  =  10.4 kDa
ORF:  432 .. 1091  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  1602 .. 1874  =  273 bp
ORF:  90 amino acids  =  9.7 kDa
ORF:  2469 .. 2732  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3321 .. 3869  =  549 bp
ORF:  182 amino acids  =  19.6 kDa
ORF:  2489 .. 3448  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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Individual Sequences & Maps

p11
pMCSG19B (linearized)
pMCSG41
pMCSG68 (linearized)
p15Tv-L
pMCSG19C
pMCSG41 (linearized)
pMCSG69
p283
pMCSG19C (linearized)
pMCSG42
pMCSG69 (linearized)
pBEN1-SGC
pMCSG20
pMCSG42 (linearized)
pMCSG7
pBEN1-SGC (linearized)
pMCSG20 (linearized)
pMCSG43
pMCSG7 (linearized)
pDB-HisGST
pMCSG21
pMCSG43 (linearized)
pMCSG70
pDB.GST
pMCSG21 (linearized)
pMCSG46
pMCSG70 (linearized)
pDB.His.MBP
pMCSG22
pMCSG46 (linearized)
pMCSG71
pDB.His.NusA
pMCSG22 (linearized)
pMCSG48
pMCSG71 (linearized)
pDB.His.TRX
pMCSG23
pMCSG48 (linearized)
pMCSG73
pDB.TRX
pMCSG23 (linearized)
pMCSG49
pMCSG73 (linearized)
pEU-GST-FV
pMCSG24
pMCSG49 (linearized)
pMCSG75
pEU-His-FV
pMCSG24 (linearized)
pMCSG50
pMCSG75 (linearized)
pEU-HSBC
pMCSG25
pMCSG50 (linearized)
pMCSG76
pFB-LIC-Bse
pMCSG25 (linearized)
pMCSG51
pMCSG76 (linearized)
pFB-LIC-Bse (linearized)
pMCSG26
pMCSG51 (linearized)
pMCSG77
pH3c-LIC
pMCSG26 (linearized)
pMCSG52
pMCSG77 (linearized)
pH3c-LIC (linearized)
pMCSG27
pMCSG52 (linearized)
pMCSG79
pHM3C-LIC
pMCSG27 (linearized)
pMCSG53
pMCSG79 (linearized)
pHM3C-LIC (linearized)
pMCSG28
pMCSG53 (linearized)
pMCSG8
pLIC-CH
pMCSG28 (linearized)
pMCSG54
pMCSG8 (linearized)
pLIC-CH (linearized)
pMCSG29
pMCSG54 (linearized)
pMCSG81
pLIC-EGFP
pMCSG29 (linearized)
pMCSG55
pMCSG81 (linearized)
pLIC-EGFP (linearized)
pMCSG30
pMCSG55 (linearized)
pMCSG9
pLIC-SGC1
pMCSG30 (linearized)
pMCSG56
pMCSG9 (linearized)
pLIC-SGC1 (linearized)
pMCSG31
pMCSG56 (linearized)
pNH-TrxT
pMCentr1
pMCSG31 (linearized)
pMCSG57
pNH-TrxT (linearized)
pMCentr1 (linearized)
pMCSG32
pMCSG57 (linearized)
pNIC-CH
pMCentr2
pMCSG32 (linearized)
pMCSG58
pNIC-CH (linearized)
pMCentr2 (linearized)
pMCSG33
pMCSG58 (linearized)
pNIC-CTHF
pMCentr3
pMCSG33 (linearized)
pMCSG59
pNIC-CTHF (linearized)
pMCentr3 (linearized)
pMCSG34
pMCSG59 (linearized)
pNIC-Zb
pMCSG10
pMCSG34 (linearized)
pMCSG60
pNIC-Zb (linearized)
pMCSG10 (linearized)
pMCSG34B
pMCSG60 (linearized)
pNIC28-Bsa4
pMCSG11
pMCSG34B (linearized)
pMCSG61
pNIC28-Bsa4 (linearized)
pMCSG11 (linearized)
pMCSG35
pMCSG61 (linearized)
pNYCOMPS-LIC-FH10T+ (N term) - linearized
pMCSG12
pMCSG35 (linearized)
pMCSG62
pNYCOMPS-LIC-FH10T+ (N term)
pMCSG12 (linearized)
pMCSG35B
pMCSG62 (linearized)
pSpeedET
pMCSG13
pMCSG35B (linearized)
pMCSG63
pVP13
pMCSG13 (linearized)
pMCSG37
pMCSG63 (linearized)
pVP16
pMCSG14
pMCSG37 (linearized)
pMCSG64
pVP33A
pMCSG14 (linearized)
pMCSG38
pMCSG64 (linearized)
pVP33K
pMCSG17
pMCSG38 (linearized)
pMCSG65
pVP56A
pMCSG17 (linearized)
pMCSG38C
pMCSG65 (linearized)
pVP56K
pMCSG18
pMCSG38C (linearized)
pMCSG66
pVP65K
pMCSG18 (linearized)
pMCSG39
pMCSG66 (linearized)
pVP68K
pMCSG19
pMCSG39 (linearized)
pMCSG67
pVP81K
pMCSG19 (linearized)
pMCSG40
pMCSG67 (linearized)
pVP91A
pMCSG19B
pMCSG40 (linearized)
pMCSG68

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Lucigen Vectors
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Yeast Plasmids
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Mammalian Expression Vectors
Structural Genomics Vectors

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