pLATE51 (linearized)

Vector for ligation independent cloning (LIC) and tightly regulated bacterial expression of a protein with an enterokinase-cleavable N-terminal 6xHis tag.

Sequence Author: Thermo Fisher

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HomePlasmidsBasic Cloning VectorspLATE51 (linearized)
rrnB T2 terminator rrnB T1 terminator AatII (4004) ZraI (4002) SspI (3886) ScaI (3562) PvuI (3452) FspI (3304) BglI (3202) AhdI (3082) AlwNI (2605) PciI (2189) BspQI - SapI (2073) NdeI (2012) BstZ17I (1962) AccI (1961) BsaAI (1943) PflFI - Tth111I (1936) PfoI (1833) AfeI (1445) BsaBI * (1388) LIC reverse sequencing primer (4297 .. 4312) lac operator Eco53kI (4315) SacI (4317) Acc65I (4319) KpnI (4323) MauBI (4330) BfuAI - BspMI - PaqCI (4335) SbfI (4346) AscI (4351) LIC forward sequencing primer (4360 .. 4379) T7 promoter LIC reverse sequencing primer (4387 .. 4401) XbaI (4407) SwaI (4432) RBS ATG NheI (4491) BmtI (4495) T7 tag (gene 10 leader) End (4526) Start (0) PmeI (18) StuI (25) SphI (33) PacI (55) LIC reverse sequencing primer (62 .. 85) HindIII (123) BspDI - ClaI (130) BamHI (157) StyI (173) T7 terminator PflMI (228) MluI (646) BclI * (660) BstEII (827) PspOMI (853) ApaI (857) ApoI (921) EcoRV (1096) HpaI (1152) KasI (1285) NarI * (1286) SfoI (1287) PluTI (1289) BspEI * (1380) pLATE51 4526 bp
AatII  (4004)
1 site		 G A C G T C C T G C A G
ZraI  (4002)
1 site		 G A C G T C C T G C A G
SspI  (3886)
1 site		 A A T A T T T T A T A A
ScaI  (3562)
1 site		 A G T A C T T C A T G A
PvuI  (3452)
1 site		 C G A T C G G C T A G C
FspI  (3304)
1 site		 T G C G C A A C G C G T
BglI  (3202)
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.
AhdI  (3082)
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.
AlwNI  (2605)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
PciI  (2189)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2073)
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  (2073)
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.
NdeI  (2012)
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.
BstZ17I  (1962)
1 site		 G T A T A C C A T A T G
AccI  (1961)
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.
BsaAI  (1943)
1 site		 Y A C G T R R T G C A Y
PflFI  (1936)
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  (1936)
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.
PfoI  (1833)
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.
AfeI  (1445)
1 site		 A G C G C T T C G C G A
BsaBI  (1388)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
Eco53kI  (4315)
1 site		 G A G C T C C T C G A G
SacI  (4317)
1 site		 G A G C T C C T C G A G
Acc65I  (4319)
1 site		 G G T A C C C C A T G G
KpnI  (4323)
1 site		 G G T A C C C C A T G G
MauBI  (4330)
1 site		 C G C G C G C G G C G C G C G C
BfuAI  (4335)
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  (4335)
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.
PaqCI  (4335)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
SbfI  (4346)
1 site		 C C T G C A G G G G A C G T C C
AscI  (4351)
1 site		 G G C G C G C C C C G C G C G G
XbaI  (4407)
1 site		 T C T A G A A G A T C T
SwaI  (4432)
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.
NheI  (4491)
1 site		 G C T A G C C G A T C G
BmtI  (4495)
1 site		 G C T A G C C G A T C G
End  (4526)
0 sites
Start  (0)
0 sites
PmeI  (18)
1 site		 G T T T A A A C C A A A T T T G
StuI  (25)
1 site		 A G G C C T T C C G G A
SphI  (33)
1 site		 G C A T G C C G T A C G
PacI  (55)
1 site		 T T A A T T A A A A T T A A T T
HindIII  (123)
1 site		 A A G C T T T T C G A A
BspDI  (130)
1 site		 A T C G A T T A G C T A
ClaI  (130)
1 site		 A T C G A T T A G C T A
BamHI  (157)
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.
StyI  (173)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
PflMI  (228)
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.
MluI  (646)
1 site		 A C G C G T T G C G C A
BclI  (660)
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  (827)
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.
PspOMI  (853)
1 site		 G G G C C C C C C G G G
ApaI  (857)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
ApoI  (921)
1 site		 R A A T T Y Y T T A A R
ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRV  (1096)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
HpaI  (1152)
1 site		 G T T A A C C A A T T G
KasI  (1285)
1 site		 G G C G C C C C G C G G
NarI  (1286)
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.
SfoI  (1287)
1 site		 G G C G C C C C G C G G
PluTI  (1289)
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.
BspEI  (1380)
1 site		 T C C G G A A G G C C T
* Blocked by Dam methylation.
LIC reverse sequencing primer
24-mer  /  46% GC
3 binding sites
4297 .. 4312  =  16 annealed bases (38% GC)
Tm  =  48°C
LIC forward sequencing primer
20-mer  /  40% GC
1 binding site
4360 .. 4379  =  20 annealed bases
Tm  =  49°C
LIC reverse sequencing primer
24-mer  /  46% GC
3 binding sites
4387 .. 4401  =  15 annealed bases (40% GC)
Tm  =  47°C
LIC reverse sequencing primer
24-mer  /  46% GC
3 binding sites
62 .. 85  =  24 annealed bases
Tm  =  58°C
lacI
296 .. 1378  =  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
296 .. 1378  =  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).
AmpR
3009 .. 3869  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   3009 .. 3800  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3009 .. 3869  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   3801 .. 3869  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3009 .. 3869  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2250 .. 2838  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2250 .. 2838  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rop
1631 .. 1822  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
1631 .. 1822  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
3870 .. 3974  =  105 bp
AmpR promoter
3870 .. 3974  =  105 bp
rrnB T1 terminator
4075 .. 4156  =  82 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4075 .. 4156  =  82 bp
transcription terminator T1 from the E. coli rrnB gene
lacI promoter
218 .. 295  =  78 bp
lacI promoter
218 .. 295  =  78 bp
ATG
4455 .. 4457  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4455 .. 4457  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
4467 .. 4484  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4467 .. 4484  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
T7 tag (gene 10 leader)
4488 .. 4520  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene 10
promotes efficient translation in E. coli
T7 tag (gene 10 leader)
4488 .. 4520  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene 10
promotes efficient translation in E. coli
T7 terminator
162 .. 209  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
162 .. 209  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
tet promoter
119 .. 147  =  29 bp
3 segments
   Segment 3:  -10  
   119 .. 124  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
119 .. 147  =  29 bp
3 segments
   Segment 2:  
   125 .. 141  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
119 .. 147  =  29 bp
3 segments
   Segment 1:  -35  
   142 .. 147  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
119 .. 147  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
rrnB T2 terminator
4248 .. 4275  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
4248 .. 4275  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
lac operator
4379 .. 4403  =  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
4379 .. 4403  =  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
4360 .. 4378  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4360 .. 4378  =  19 bp
promoter for bacteriophage T7 RNA polymerase
lac operator
4293 .. 4309  =  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
4293 .. 4309  =  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).
RBS
4440 .. 4446  =  7 bp
ribosome binding site
RBS
4440 .. 4446  =  7 bp
ribosome binding site
ORF:  229 .. 546  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  1135 .. 1527  =  393 bp
ORF:  130 amino acids  =  14.0 kDa
ORF:  3139 .. 3405  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  419 .. 1378  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  1598 .. 1822  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  1243 .. 1599  =  357 bp
ORF:  118 amino acids  =  13.4 kDa
ORF:  3009 .. 3869  =  861 bp
ORF:  286 amino acids  =  31.5 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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