pF25K ICE T7

Flexi® vector with a kanamycin resistance marker, for cell-free translation in an insect cell extract.

Sequence Author: Promega

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HomePlasmidsBasic Cloning VectorspF25K ICE T7
AseI (20) AclI (3782) BsaI (3646) ScaI (3459) MluI (3375) BspEI (3162) BspHI (3118) ApaLI (2712) PciI (2398) AgeI (2330) BstBI (2303) RsrII (2137) NaeI (2123) NgoMIV (2121) T7 promoter AsiSI - PvuI (90) BtgI - NcoI (93) PmeI (457) EcoRI (462) Acc65I (474) KpnI (478) BamHI (483) XbaI (489) SalI (495) AccI (496) HincII (497) SbfI (505) BlpI (570) AfeI (645) MfeI (676) SnaBI (687) PsiI (712) SwaI (777) EcoO109I (1034) NruI (1069) NotI (1074) StuI (1083) BtsI - BtsαI (1185) BstAPI (1319) XcmI (1437) BclI * (1462) MscI (1703) FspI (1723) PflFI - Tth111I (1739) BsrDI (1854) pF25K ICE T7 3942 bp
AseI  (20)
1 site		 A T T A A T T A A T T A
AclI  (3782)
1 site		 A A C G T T T T G C A A
BsaI  (3646)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
ScaI  (3459)
1 site		 A G T A C T T C A T G A
MluI  (3375)
1 site		 A C G C G T T G C G C A
BspEI  (3162)
1 site		 T C C G G A A G G C C T
BspHI  (3118)
1 site		 T C A T G A A G T A C T
ApaLI  (2712)
1 site		 G T G C A C C A C G T G
PciI  (2398)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AgeI  (2330)
1 site		 A C C G G T T G G C C A
BstBI  (2303)
1 site		 T T C G A A A A G C T T
RsrII  (2137)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
NaeI  (2123)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (2121)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
AsiSI  (90)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (90)
1 site		 C G A T C G G C T A G C
BtgI  (93)
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  (93)
1 site		 C C A T G G G G T A C C
PmeI  (457)
1 site		 G T T T A A A C C A A A T T T G
EcoRI  (462)
1 site		 G A A T T C C T T A A G
Acc65I  (474)
1 site		 G G T A C C C C A T G G
KpnI  (478)
1 site		 G G T A C C C C A T G G
BamHI  (483)
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.
XbaI  (489)
1 site		 T C T A G A A G A T C T
SalI  (495)
1 site		 G T C G A C C A G C T G
AccI  (496)
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.
HincII  (497)
1 site		 G T Y R A C C A R Y T G
SbfI  (505)
1 site		 C C T G C A G G G G A C G T C C
BlpI  (570)
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.
AfeI  (645)
1 site		 A G C G C T T C G C G A
MfeI  (676)
1 site		 C A A T T G G T T A A C
SnaBI  (687)
1 site		 T A C G T A A T G C A T
PsiI  (712)
1 site		 T T A T A A A A T A T T
SwaI  (777)
1 site		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
EcoO109I  (1034)
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.
NruI  (1069)
1 site		 T C G C G A A G C G C T
NotI  (1074)
1 site		 G C G G C C G C C G C C G G C G
StuI  (1083)
1 site		 A G G C C T T C C G G A
BtsI  (1185)
1 site		 G C A G T G N N C G T C A C
BtsαI  (1185)
1 site		 G C A G T G N N C G T C A C
Sticky ends from different BtsαI sites may not be compatible.
BstAPI  (1319)
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.
XcmI  (1437)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BclI  (1462)
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.
MscI  (1703)
1 site		 T G G C C A A C C G G T
FspI  (1723)
1 site		 T G C G C A A C G C G T
PflFI  (1739)
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  (1739)
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.
BsrDI  (1854)
1 site		 G C A A T G N N C G T T A C
Sticky ends from different BsrDI sites may not be compatible.
NeoR/KanR
1493 .. 2287  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1493 .. 2287  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
2459 .. 3047  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2459 .. 3047  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
polyhedrin 3' UTR
611 .. 983  =  373 bp
3' UTR from the Autographa californica nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)
polyhedrin 3' UTR
611 .. 983  =  373 bp
3' UTR from the Autographa californica nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)
barnase
117 .. 452  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
117 .. 452  =  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
3163 .. 3446  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3163 .. 3446  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
rrnB T1 terminator
3676 .. 3762  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3676 .. 3762  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
1018 .. 1065  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1018 .. 1065  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
polyhedrin 5' UTR
40 .. 85  =  46 bp
5' UTR from the Malacosoma neustria nucleopolyhedrovirus polyhedrin gene (Ezure et al., 2006)
polyhedrin 5' UTR
40 .. 85  =  46 bp
5' UTR from the Malacosoma neustria nucleopolyhedrovirus polyhedrin gene (Ezure et al., 2006)
poly(A)
988 .. 1017  =  30 bp
synthetic poly(A) tract
poly(A)
988 .. 1017  =  30 bp
synthetic poly(A) tract
rrnB T2 terminator
3854 .. 3881  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
3854 .. 3881  =  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:  1493 .. 2287  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  117 .. 452  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  1665 .. 2051  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  3408 .. 3755  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  3517 .. 3843  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  1802 .. 2056  =  255 bp
ORF:  84 amino acids  =  9.8 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

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I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
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TOPO Cloning Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
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Mammalian Expression Vectors
Structural Genomics Vectors

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