pMMB66EH

Broad-host-range bacterial cloning vector. See also pMMB66HE.
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HomePlasmidsBasic Cloning VectorspMMB66EH
EcoRI (8806) lac operator MauBI (8531) MluI (8074) ApaI - BanII (7871) PspOMI (7867) HpaI (7572) BsmBI - Esp3I (7459) BspQI - SapI (7266) FspAI (6795) BsaAI (6740) EcoNI (6645) PfoI * (6505) SgrAI (4557) TspMI - XmaI (3) SmaI (5) BamHI (8) SalI (14) PstI (24) HindIII (28) rrnB T2 terminator BssSI - BssSαI (670) DraI (761) PvuI (970) AhdI (1339) CsiI - SexAI * (1463) SfiI (1697) EcoO109I (2249) BstZ17I (3072) PasI (4078) pMMB66EH 8808 bp
EcoRI  (8806)
1 site		 G A A T T C C T T A A G
MauBI  (8531)
1 site		 C G C G C G C G G C G C G C G C
MluI  (8074)
1 site		 A C G C G T T G C G C A
ApaI  (7871)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BanII  (7871)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
PspOMI  (7867)
1 site		 G G G C C C C C C G G G
HpaI  (7572)
1 site		 G T T A A C C A A T T G
BsmBI  (7459)
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  (7459)
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.
BspQI  (7266)
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  (7266)
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.
FspAI  (6795)
1 site		 R T G C G C A Y Y A C G C G T R
BsaAI  (6740)
1 site		 Y A C G T R R T G C A Y
EcoNI  (6645)
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  (6505)
1 site		 T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
SgrAI  (4557)
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.
TspMI  (3)
1 site		 C C C G G G G G G C C C
XmaI  (3)
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  (5)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BamHI  (8)
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.
SalI  (14)
1 site		 G T C G A C C A G C T G
PstI  (24)
1 site		 C T G C A G G A C G T C
HindIII  (28)
1 site		 A A G C T T T T C G A A
BssSI  (670)
1 site		 C A C G A G G T G C T C
BssSαI  (670)
1 site		 C A C G A G G T G C T C
DraI  (761)
1 site		 T T T A A A A A A T T T
PvuI  (970)
1 site		 C G A T C G G C T A G C
AhdI  (1339)
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.
CsiI  (1463)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (1463)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (1697)
1 site		 G G C C N N N N N G G C C C C G G N N N N N C C G G
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
EcoO109I  (2249)
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.
BstZ17I  (3072)
1 site		 G T A T A C C A T A T G
PasI  (4078)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
lacI
7347 .. 8429  =  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
7347 .. 8429  =  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).
RSF1010 RepB
3911 .. 4882  =  972 bp
323 amino acids  =  35.9 kDa
Product: replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepB
3911 .. 4882  =  972 bp
323 amino acids  =  35.9 kDa
Product: replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
AmpR
552 .. 1412  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   552 .. 620  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
552 .. 1412  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   621 .. 1412  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
552 .. 1412  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
RSF1010 RepC
6219 .. 7070  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepC
6219 .. 7070  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriV
1849 .. 2243  =  395 bp
replication origin of the broad-host-range plasmid RSF1010; requires the RSF1010 RepA/B/C proteins for replication (Scholz et al., 1989)
RSF1010 oriV
1849 .. 2243  =  395 bp
replication origin of the broad-host-range plasmid RSF1010; requires the RSF1010 RepA/B/C proteins for replication (Scholz et al., 1989)
AmpR promoter
460 .. 551  =  92 bp
AmpR promoter
460 .. 551  =  92 bp
RSF1010 oriT
2585 .. 2672  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriT
2585 .. 2672  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
rrnB T1 terminator
236 .. 322  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
236 .. 322  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
lacIq promoter
8430 .. 8507  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
8430 .. 8507  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
MCS
8806 .. 33  =  36 bp
multiple cloning site
MCS
8806 .. 33  =  36 bp
multiple cloning site
tac promoter
8737 .. 8765  =  29 bp
3 segments
   Segment 1:  -35  
   8737 .. 8742  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8737 .. 8765  =  29 bp
3 segments
   Segment 2:  
   8743 .. 8758  =  16 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8737 .. 8765  =  29 bp
3 segments
   Segment 3:  -10  
   8759 .. 8765  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
8737 .. 8765  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
rrnB T2 terminator
414 .. 441  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
414 .. 441  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
lac operator
8773 .. 8789  =  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
8773 .. 8789  =  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).
RSF1010 RepA
5393 .. 6232  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepA
5393 .. 6232  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
ORF:  1510 .. 1842  =  333 bp
ORF:  110 amino acids  =  12.4 kDa
ORF:  2753 .. 4882  =  2130 bp
ORF:  709 amino acids  =  78.0 kDa
ORF:  5393 .. 6232  =  840 bp
ORF:  279 amino acids  =  29.9 kDa
ORF:  552 .. 1412  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2049 .. 2477  =  429 bp
ORF:  142 amino acids  =  15.3 kDa
ORF:  2595 .. 3002  =  408 bp
ORF:  135 amino acids  =  14.3 kDa
ORF:  3501 .. 3914  =  414 bp
ORF:  137 amino acids  =  15.1 kDa
ORF:  4338 .. 4982  =  645 bp
ORF:  214 amino acids  =  22.8 kDa
ORF:  6120 .. 7070  =  951 bp
ORF:  316 amino acids  =  34.4 kDa
ORF:  1399 .. 1716  =  318 bp
ORF:  105 amino acids  =  12.2 kDa
ORF:  2920 .. 3150  =  231 bp
ORF:  76 amino acids  =  8.4 kDa
ORF:  3817 .. 4170  =  354 bp
ORF:  117 amino acids  =  12.6 kDa
ORF:  5644 .. 6009  =  366 bp
ORF:  121 amino acids  =  13.3 kDa
ORF:  7327 .. 7590  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  8179 .. 8496  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  1431 .. 1709  =  279 bp
ORF:  92 amino acids  =  10.3 kDa
ORF:  6453 .. 6791  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
ORF:  7347 .. 8306  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  77 .. 403  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  1016 .. 1282  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2270 .. 2554  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  2624 .. 3280  =  657 bp
ORF:  218 amino acids  =  22.9 kDa
ORF:  3473 .. 3811  =  339 bp
ORF:  112 amino acids  =  12.0 kDa
ORF:  4028 .. 4621  =  594 bp
ORF:  197 amino acids  =  20.5 kDa
ORF:  5390 .. 6157  =  768 bp
ORF:  255 amino acids  =  27.0 kDa
ORF:  6560 .. 6901  =  342 bp
ORF:  113 amino acids  =  12.6 kDa
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Individual Sequences & Maps

BlueScribe
pEASY-T1 (linearized)
pJET1.2
pSMART LCAmp
CAG promoter
pEASY-T1 Simple (linearized)
pJET1.2 blunt
pSMART LCAmp (linearized)
chicken beta-actin promoter
pEASY-T3 (linearized)
pJW168
pSMART LCKan
CLIP-tag
pEASY-T5 Zero (linearized)
pKF 18k-2
pSMART LCKan (linearized)
CMV promoter
pEX-1
pKF 19k-2
pSP64 Poly(A)
GAL1 promoter
pEX-A
pKOV
pSP72
GST
pEX-C-GST
pLATE11 (linearized)
pSP73
HaloTag
pEX-C-His
pLATE31 (linearized)
pSpark Done (linearized)
HSV TK promoter
pEX-K
pLATE51 (linearized)
pSpark I (linearized)
lac operator
pEX-N-GST
pLATE52 (linearized)
pSpark II (linearized)
lac promoter
pEX-N-His
pLEXX-AK
pSpark III (linearized)
lac UV5 promoter
pEX-N-His-GST
pLPS-B (linearized)
pSpark IV (linearized)
lacZ
pEX1
pMAL-c5E
pSpark V (linearized)
lambda
pEX2
pMAL-c5G
pSpot1
LITMUS28
pEX3
pMAL-c5X
pSpot2
LITMUS28i
pEZSeq-Amp
pMAL-c5X-His
pSpot3
LITMUS29
pEZSeq-Amp (linearized)
pMAL-c6T
pSpot4
LITMUS38
pEZSeq-Kan
pMAL-p5E
pSTBlue-1
LITMUS38i
pEZSeq-Kan (linearized)
pMAL-p5G
pSTV28
LITMUS39
pEZ BAC
pMAL-p5X
pSTV29
M13mp18
pEZ BAC (BamHI linearized)
pMCS5
pT7Blue
M13mp19
pEZ BAC (HpaI linearized)
pMiniT
pT7Blue-2
MBP
pF1A T7
pMiniT (linearized)
pT7Blue-3
pACYC177
pF1K T7
pMiniT 2.0
pT7CFE1-CFtag
pACYC184
pF25A ICE T7
pMiniT 2.0 (linearized)
pT7CFE1-CGFP-HA-His
pAN7-1
pF25K ICE T7
pMMB190
pT7CFE1-CGST-HA-His
pANT7_cGST
pF3A WG (BYDV)
pMMB206
pT7CFE1-CHA
pANT7_nHA
pF3K WG (BYDV)
pMMB207
pT7CFE1-CHis
pAUR316
pFC20A HaloTag T7 SP6
pMMB208
pT7CFE1-CMyc
pBC KS(+)
pFC20K HaloTag T7 SP6
pMMB66EH
pT7CFE1-NFtag
pBC KS(-)
pFC30A His6HaloTag T7
pMMB66HE
pT7CFE1-NHA
pBC SK(+)
pFC30K His6HaloTag T7
pMMB67EH
pT7CFE1-NHA-CHis
pBC SK(-)
pFC7A (HQ)
pMMB67HE
pT7CFE1-NHis
pBeloBAC11
pFC7K (HQ)
pNEB193
pT7CFE1-NHis-GST
pBluescript II KS(+)
pFN18A HaloTag T7
pNEB206A (linearized)
pT7CFE1-NHis-GST-CHA
pBluescript II KS(-)
pFN18K HaloTag T7
pOSIP-CH
pT7CFE1-NMyc
pBluescript II SK(+)
pFN19A HaloTag T7 SP6
pOSIP-CO
pTOP Blunt V2 (linearized)
pBluescript II SK(-)
pFN19K HaloTag T7 SP6
pOSIP-CT
pTrcHis A
pBluescript KS(+)
pFN29A His6HaloTag T7
pOSIP-KC
pTrcHis B
pBluescript KS(-)
pFN29K His6HaloTag T7
pOSIP-KH
pTrcHis C
pBluescript SK(+)
pFN2A (GST)
pOSIP-KL
pTrcHis2 A
pBluescript SK(-)
pFN2K (GST)
pOSIP-KO
pTrcHis2 B
pBR322
pFN6A (HQ)
pOSIP-KP
pTrcHis2 C
pBS KS(+)
pFN6K (HQ)
pOSIP-KT
pTV 118N
pBS KS(-)
pGEM-11Zf(+)
pOSIP-TH
pTWV228
pBS SK(+)
pGEM-11Zf(-)
pOSIP-TO
pTXB1
pBS SK(-)
pGEM-3Z
pOSIP-TT
pTYB21
pBS(+)
pGEM-3Zf(+)
pPCR-Script Amp SK(+)
pType IIs
pBS(-)