pMCSG76

Bacterial vector with a CloDF13 origin encoding tRNA genes for rare Arg and Ile codons, for expressing a protein with a 6xHis-TEV leader plus a second untagged protein.

Sequence Author: Midwest Center for Structural Genomics

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HomePlasmidsStructural Genomics VectorspMCSG76
EcoNI (4304) PfoI (4268) PflMI (4261) AclI (4176) BstAPI (4160) MluI (3836) ApaI (3633) PspOMI (3629) HpaI (3334) BsmBI - Esp3I (3221) PciI (2216) BmtI (2120) BspEI * (2) PaeR7I - PspXI - XhoI (158) EagI - NotI (166) HindIII (173) SalI (179) AccI (180) Eco53kI (188) SacI (190) EcoRI (192) TspMI - XmaI (232) SmaI (234) RBS SspI (275) TatI (287) Acc65I (300) KpnI (304) BglII (307) 6xHis ATG NdeI (343) RBS lac operator T7 promoter BspDI * - ClaI * (450) AgeI (492) XmnI (561) AscI (709) PstI - SbfI (720) BstBI (725) MreI (857) AfeI (943) NcoI (1127) PsiI (1142) PflFI - Tth111I (1176) BsaAI (1265) BglI (1393) PvuI (1643) NheI (2116) pMCSG76 4310 bp
EcoNI  (4304)
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  (4268)
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  (4261)
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.
AclI  (4176)
1 site		 A A C G T T T T G C A A
BstAPI  (4160)
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.
MluI  (3836)
1 site		 A C G C G T T G C G C A
ApaI  (3633)
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  (3629)
1 site		 G G G C C C C C C G G G
HpaI  (3334)
1 site		 G T T A A C C A A T T G
BsmBI  (3221)
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  (3221)
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.
PciI  (2216)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BmtI  (2120)
1 site		 G C T A G C C G A T C G
BspEI  (2)
1 site		 T C C G G A A G G C C T
* Blocked by Dam methylation.
PaeR7I  (158)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (158)
1 site		 C T C G A G G A G C T C
EagI  (166)
1 site		 C G G C C G G C C G G C
NotI  (166)
1 site		 G C G G C C G C C G C C G G C G
HindIII  (173)
1 site		 A A G C T T T T C G A A
SalI  (179)
1 site		 G T C G A C C A G C T G
AccI  (180)
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.
Eco53kI  (188)
1 site		 G A G C T C C T C G A G
SacI  (190)
1 site		 G A G C T C C T C G A G
EcoRI  (192)
1 site		 G A A T T C C T T A A G
TspMI  (232)
1 site		 C C C G G G G G G C C C
XmaI  (232)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (234)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
SspI  (275)
1 site		 A A T A T T T T A T A A
TatI  (287)
1 site		 W G T A C W W C A T G W
Acc65I  (300)
1 site		 G G T A C C C C A T G G
KpnI  (304)
1 site		 G G T A C C C C A T G G
BglII  (307)
1 site		 A G A T C T T C T A G A
NdeI  (343)
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.
BspDI  (450)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (450)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
AgeI  (492)
1 site		 A C C G G T T G G C C A
XmnI  (561)
1 site		 G A A N N N N T T C C T T N N N N A A G
AscI  (709)
1 site		 G G C G C G C C C C G C G C G G
PstI  (720)
1 site		 C T G C A G G A C G T C
SbfI  (720)
1 site		 C C T G C A G G G G A C G T C C
BstBI  (725)
1 site		 T T C G A A A A G C T T
MreI  (857)
1 site		 C G C C G G C G G C G G C C G C
AfeI  (943)
1 site		 A G C G C T T C G C G A
NcoI  (1127)
1 site		 C C A T G G G G T A C C
PsiI  (1142)
1 site		 T T A T A A A A T A T T
PflFI  (1176)
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  (1176)
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.
BsaAI  (1265)
1 site		 Y A C G T R R T G C A Y
BglI  (1393)
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.
PvuI  (1643)
1 site		 C G A T C G G C T A G C
NheI  (2116)
1 site		 G C T A G C C G A T C G
lacI
3109 .. 4191  =  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
3109 .. 4191  =  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).
SmR
1230 .. 2021  =  792 bp
263 amino acids  =  29.3 kDa
Product: aminoglycoside adenylyltransferase (Murphy, 1985)
confers resistance to spectinomycin and streptomycin
SmR
1230 .. 2021  =  792 bp
263 amino acids  =  29.3 kDa
Product: aminoglycoside adenylyltransferase (Murphy, 1985)
confers resistance to spectinomycin and streptomycin
CloDF13 ori
2161 .. 2899  =  739 bp
Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins.
CloDF13 ori
2161 .. 2899  =  739 bp
Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins.
AmpR promoter
2022 .. 2113  =  92 bp
AmpR promoter
2022 .. 2113  =  92 bp
lacI promoter
4192 .. 4269  =  78 bp
lacI promoter
4192 .. 4269  =  78 bp
argU
707 .. 783  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
argU
707 .. 783  =  77 bp
Product: E. coli tRNA-Arg recognizing AGA and AGG
ileX
1053 .. 1128  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
ileX
1053 .. 1128  =  76 bp
Product: E. coli tRNA-Ile recognizing AUA
TEV site
279 .. 299  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
279 .. 299  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
6xHis
324 .. 341  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
324 .. 341  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
342 .. 344  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
342 .. 344  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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
389 .. 413  =  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
389 .. 413  =  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
414 .. 432  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
414 .. 432  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
243 .. 248  =  6 bp
ribosome binding site
RBS
243 .. 248  =  6 bp
ribosome binding site
RBS
353 .. 358  =  6 bp
ribosome binding site
RBS
353 .. 358  =  6 bp
ribosome binding site
ORF:  1444 .. 1722  =  279 bp
ORF:  92 amino acids  =  9.4 kDa
ORF:  1732 .. 1989  =  258 bp
ORF:  85 amino acids  =  9.1 kDa
ORF:  842 .. 1096  =  255 bp
ORF:  84 amino acids  =  8.5 kDa
ORF:  324 .. 557  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  1599 .. 1982  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  3075 .. 3326  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  1230 .. 2021  =  792 bp
ORF:  263 amino acids  =  29.3 kDa
ORF:  3089 .. 3352  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3941 .. 4306  =  366 bp
ORF:  121 amino acids  =  13.1 kDa
ORF:  3109 .. 4068  =  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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