pDream2.1 MCS

Vector for expression of N-terminally FLAG-tagged proteins in bacterial, insect, and mammalian cells.

Sequence Author: GenScript

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HomePlasmidsInsect Cell VectorspDream2.1 MCS
SalI (243) ScaI (6790) AlwNI (5991) PciI (5576) AsiSI (5432) BmgBI (5390) SwaI (4976) AfeI (4844) BspHI (4486) BsmI (4434) BstBI (4196) RsrII (4030) BssHII (3911) PflFI - Tth111I (3632) PstI (3567) AccI (244) HincII (245) MluI (399) FseI (1017) PaeR7I - PspXI - XhoI (1020) CMV enhancer T7 primer (1615 .. 1634) lac operator PpuMI (1660) PacI (1732) attB1 ATG FLAG BamHI (1837) BglII (1844) EcoRI (1851) Acc65I (1857) MCS KpnI (1861) Eco53kI (1865) SacI (1867) HindIII (1869) NotI (1876) SP6 primer (1883 .. 1904) attB2 AleI (1988) BsgI (2172) BlpI (2258) XbaI (2456) DraIII (2781) CsiI - SexAI * (3071) BseRI (3300) StuI (3303) AvrII (3304) TspMI - XmaI (3325) SmaI (3327) BclI * (3355) BsaBI * (3373) KasI (3513) NarI (3514) SfoI (3515) PluTI (3517) pDream2.1/MCS 7159 bp
SalI  (243)
1 site		 G T C G A C C A G C T G
ScaI  (6790)
1 site		 A G T A C T T C A T G A
AlwNI  (5991)
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  (5576)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AsiSI  (5432)
1 site		 G C G A T C G C C G C T A G C G
BmgBI  (5390)
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.
SwaI  (4976)
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.
AfeI  (4844)
1 site		 A G C G C T T C G C G A
BspHI  (4486)
1 site		 T C A T G A A G T A C T
BsmI  (4434)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
BstBI  (4196)
1 site		 T T C G A A A A G C T T
RsrII  (4030)
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.
BssHII  (3911)
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.
PflFI  (3632)
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  (3632)
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.
PstI  (3567)
1 site		 C T G C A G G A C G T C
AccI  (244)
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  (245)
1 site		 G T Y R A C C A R Y T G
MluI  (399)
1 site		 A C G C G T T G C G C A
FseI  (1017)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
PaeR7I  (1020)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1020)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1020)
1 site		 C T C G A G G A G C T C
PpuMI  (1660)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
PacI  (1732)
1 site		 T T A A T T A A A A T T A A T T
BamHI  (1837)
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.
BglII  (1844)
1 site		 A G A T C T T C T A G A
EcoRI  (1851)
1 site		 G A A T T C C T T A A G
Acc65I  (1857)
1 site		 G G T A C C C C A T G G
KpnI  (1861)
1 site		 G G T A C C C C A T G G
Eco53kI  (1865)
1 site		 G A G C T C C T C G A G
SacI  (1867)
1 site		 G A G C T C C T C G A G
HindIII  (1869)
1 site		 A A G C T T T T C G A A
NotI  (1876)
1 site		 G C G G C C G C C G C C G G C G
AleI  (1988)
1 site		 C A C N N N N G T G G T G N N N N C A C
BsgI  (2172)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BlpI  (2258)
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.
XbaI  (2456)
1 site		 T C T A G A A G A T C T
DraIII  (2781)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
CsiI  (3071)
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  (3071)
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.
BseRI  (3300)
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.
StuI  (3303)
1 site		 A G G C C T T C C G G A
AvrII  (3304)
1 site		 C C T A G G G G A T C C
TspMI  (3325)
1 site		 C C C G G G G G G C C C
XmaI  (3325)
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  (3327)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BclI  (3355)
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.
BsaBI  (3373)
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.
KasI  (3513)
1 site		 G G C G C C C C G C G G
NarI  (3514)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (3515)
1 site		 G G C G C C C C G C G G
PluTI  (3517)
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.
T7 primer
20-mer  /  40% GC
1 binding site
1615 .. 1634  =  20 annealed bases
Tm  =  49°C
SP6 primer
22-mer  /  36% GC
1 binding site
1883 .. 1904  =  22 annealed bases
Tm  =  51°C
AmpR
6237 .. 7097  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6237 .. 7028  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6237 .. 7097  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7029 .. 7097  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6237 .. 7097  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
baculovirus recombination region (lef2/ORF603)
181 .. 1008  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
181 .. 1008  =  828 bp
contains ORF603 and part of lef2
NeoR/KanR
3386 .. 4180  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
3386 .. 4180  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
baculovirus recombination region (ORF1629)
4722 .. 5427  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4722 .. 5427  =  706 bp
contains part of ORF1629
ori
5637 .. 6224  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5637 .. 6224  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hGH poly(A) signal
1931 .. 2407  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1931 .. 2407  =  477 bp
human growth hormone polyadenylation signal
f1 ori
2548 .. 2976  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2548 .. 2976  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2990 .. 3319  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
2990 .. 3319  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
1086 .. 1389  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1086 .. 1389  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
1390 .. 1593  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
1390 .. 1593  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
4354 .. 4475  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4354 .. 4475  =  122 bp
SV40 polyadenylation signal
p10 promoter
1661 .. 1770  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
1661 .. 1770  =  110 bp
baculovirus promoter for expression in insect cells
β-globin poly(A) signal
4519 .. 4574  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
4519 .. 4574  =  56 bp
rabbit β-globin polyadenylation signal
T7 terminator
2408 .. 2455  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2408 .. 2455  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
MCS
1837 .. 1882  =  46 bp
multiple cloning site
MCS
1837 .. 1882  =  46 bp
multiple cloning site
ATG
1810 .. 1812  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1810 .. 1812  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
FLAG
1813 .. 1836  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG epitope tag, followed by an enterokinase cleavage site
FLAG
1813 .. 1836  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG epitope tag, followed by an enterokinase cleavage site
attB1
1772 .. 1796  =  25 bp
recombination site for the Gateway® BP reaction
attB1
1772 .. 1796  =  25 bp
recombination site for the Gateway® BP reaction
attB2
1905 .. 1929  =  25 bp
recombination site for the Gateway® BP reaction
attB2
1905 .. 1929  =  25 bp
recombination site for the Gateway® BP reaction
lac operator
1634 .. 1658  =  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
1634 .. 1658  =  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
1615 .. 1633  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1615 .. 1633  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF603
343 .. 948  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
343 .. 948  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF1629
4839 .. 5427  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4839 .. 5427  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
SV40 ori
3170 .. 3305  =  136 bp
SV40 origin of replication
SV40 ori
3170 .. 3305  =  136 bp
SV40 origin of replication
lef2
181 .. 305  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
181 .. 305  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
ORF:  6367 .. 6633  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7147 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  3386 .. 4180  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2499 .. 2765  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  3558 .. 3944  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3695 .. 4231  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  387 .. 617  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
ORF:  4839 .. 5456  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  6237 .. 7097  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

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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