pFC30A His6HaloTag T7

Flexi® vector with an ampicillin resistance marker, for bacterial or cell-free expression of a protein with a cleavable C-terminal HaloTag®-6xHis dual tag.

Sequence Author: Promega

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HomePlasmidsBasic Cloning VectorspFC30A His6HaloTag T7
T7 promoter BglII (3864) MluI (3745) BspEI (3532) PciI (2768) AgeI (2700) BstBI (2673) FspI (2375) AsiSI - PvuI - SgfI (65) BtgI - NcoI (68) Eco53kI - EcoICRI (449) SacI (451) PaeR7I - XhoI (452) BamHI (497) PflMI (538) SphI (562) AleI (596) BstEII (614) BseRI (616) PflFI - Tth111I (746) BmgBI (748) PasI (778) StuI * (947) BclI * (978) BspDI * - ClaI * (984) PshAI (1043) SgrAI (1130) BbsI (1329) BssHII (1355) AccI (1367) BlpI (1419) EcoO109I (1446) BfuAI - BspMI (1571) SnaBI (1660) SspI (1793) XmnI (1998) pFC30A His6HaloTag® T7 4312 bp
BglII  (3864)
1 site		 A G A T C T T C T A G A
MluI  (3745)
1 site		 A C G C G T T G C G C A
BspEI  (3532)
1 site		 T C C G G A A G G C C T
PciI  (2768)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AgeI  (2700)
1 site		 A C C G G T T G G C C A
BstBI  (2673)
1 site		 T T C G A A A A G C T T
FspI  (2375)
1 site		 T G C G C A A C G C G T
AsiSI  (65)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (65)
1 site		 C G A T C G G C T A G C
SgfI  (65)
1 site		 G C G A T C G C C G C T A G C G
BtgI  (68)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
NcoI  (68)
1 site		 C C A T G G G G T A C C
Eco53kI  (449)
1 site		 G A G C T C C T C G A G
EcoICRI  (449)
1 site		 G A G C T C C T C G A G
SacI  (451)
1 site		 G A G C T C C T C G A G
PaeR7I  (452)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (452)
1 site		 C T C G A G G A G C T C
BamHI  (497)
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.
PflMI  (538)
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.
SphI  (562)
1 site		 G C A T G C C G T A C G
AleI  (596)
1 site		 C A C N N N N G T G G T G N N N N C A C
BstEII  (614)
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.
BseRI  (616)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
PflFI  (746)
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  (746)
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.
BmgBI  (748)
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.
PasI  (778)
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.
StuI  (947)
1 site		 A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (978)
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.
BspDI  (984)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (984)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (1043)
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.
SgrAI  (1130)
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.
BbsI  (1329)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BssHII  (1355)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
AccI  (1367)
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.
BlpI  (1419)
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  (1446)
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.
BfuAI  (1571)
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  (1571)
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.
SnaBI  (1660)
1 site		 T A C G T A A T G C A T
SspI  (1793)
1 site		 A A T A T T T T A T A A
XmnI  (1998)
1 site		 G A A N N N N T T C C T T N N N N A A G
TEV site
467 .. 487  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
467 .. 487  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
HaloTag®
503 .. 1384  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
HaloTag®
503 .. 1384  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
6xHis
1388 .. 1405  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1388 .. 1405  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
AmpR
1811 .. 2671  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1811 .. 1879  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1811 .. 2671  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1880 .. 2671  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1811 .. 2671  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2829 .. 3417  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2829 .. 3417  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
92 .. 427  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
92 .. 427  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
cer region
3533 .. 3816  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3533 .. 3816  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
AmpR promoter
1706 .. 1810  =  105 bp
AmpR promoter
1706 .. 1810  =  105 bp
rrnB T1 terminator
4046 .. 4132  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4046 .. 4132  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
1430 .. 1477  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1430 .. 1477  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
rrnB T2 terminator
4224 .. 4251  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
4224 .. 4251  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  3778 .. 4125  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  92 .. 427  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  434 .. 1408  =  975 bp
ORF:  324 amino acids  =  36.8 kDa
ORF:  1811 .. 2671  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  431 .. 1399  =  969 bp
ORF:  322 amino acids  =  36.1 kDa
ORF:  3887 .. 4213  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  1060 .. 1296  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2275 .. 2541  =  267 bp
ORF:  88 amino acids  =  9.2 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(-)
pGEM-3Zf(-)
pPCR-Script Cam SK(+)
pTZ19R
pBSK(+) Simple-Amp
pGEM-4Z
pPTR I
pTZ19U
pBSK(+) Simple-Kan
pGEM-5Zf(+)
pPTR II
pTZ57R
pcDNA3
pGEM-7Zf(+)
pRACE (linearized)
pUAST
pcDNA3.1(+)
pGEM-7Zf(-)
pRANGER-BTB-1
pUC118
pcDNA3.1(-)
pGEM-9Zf(-)
pRANGER-BTB-2
pUC119
pCold I
pGEM-T
pRANGER-BTB-3
pUC18
pCold II
pGEM-T (linearized)
pRANGER-BTB-5
pUC19
pCold III
pGEM-T Easy
pRHA-109
pUC19L
pCold IV
pGEM-T Easy (linearized)
pRHA-113
pUC57
pCold TF
pGGA
pRHA-67
pUC57-Kan
pCR-XL-2-TOPO
pGGAselect
pRSET A
pUC57-Simple
pCR-XL-2-TOPO (linearized)
PGK promoter
pRSET B
pUCmu
pDD
pGS-21a
pRSET C
pVLT31
pDNR-Dual
pH6HTC His6HaloTag T7
pSB1C3
pVLT33
pDNR-LIB
pH6HTN His6HaloTag T7
pSC101
pVLT35
pE-SUMOpro Amp
pHP45(omega)
pSEAP2-Basic
pYES1L
pE-SUMOpro Kan
pHP45(omega)-Km
pSF-Core
pZErO-2
pE-SUMOpro3 Amp
pHP45(omega)-Tc
pSMART BAC v2.0
SNAP-tag
pE-SUMOpro3 Kan
pHSG298
pSMART BAC v2.0 (BamHI linearized)
SP6 promoter
pE-SUMOstar Amp
pHSG299
pSMART BAC v2.0 (EcoRI linearized)
SV40 promoter
pE-SUMOstar Kan
pHSG396
pSMART BAC v2.0 (HindIII linearized)
T3 promoter
pEASY-Blunt (linearized)
pHSG398
pSMART FOS v2.0 (linearized)
T5 promoter
pEASY-Blunt E1 (linearized)
pHY300PLK
pSMART HCAmp
T7 promoter
pEASY-Blunt E2 (linearized)
pICOz
pSMART HCAmp (linearized)
tac promoter
pEASY-Blunt Simple (linearized)
pInvRecA
pSMART HCKan
trc promoter
pEASY-Blunt Zero (linearized)
pJAZZ-OC
pSMART HCKan (linearized)
phiX174
pEASY-Blunt3 (linearized)
pJAZZ-OK

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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