Resources
Plasmid Files

pZIP-mEF1-alpha-RFP-Puro

Lentiviral vector with a mouse EF-1α promoter-driven cassette encoding TurboRFP plus a short hairpin RNA (shRNA) with an "UltramiR" microRNA scaffold.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pZIP-mEF1-alpha-RFP-Puro Sequence and MappZIP-mEF1-alpha-RFP-Puro.dna
Map and Sequence File   
Sequence Author:  transOMIC
Download Free Trial Get SnapGene Viewer

 SgrDI (9112) SspI (8995) PvuI (8561) FspI (8413) AhdI (8191) PmeI (6933) PsiI (6883) PstI (6424) AscI (5806) MluI (5799) HpaI (5558) BamHI (5472) NotI (5462) BsmBI (5253) BstEII (4995) RsrII (4977) BsiWI (4917) PflFI - Tth111I (4903) BmgBI (4826) PmlI (4599) NdeI (276) SnaBI (382) NheI (405) BmtI (409) HIV-1 Ψ NruI * (867) BspQI - SapI (1136) FseI (1184) MfeI (1223) BbvCI (1458) KflI (1968) BspDI - ClaI (2214) BlpI (2344) BstXI (3003) XcmI (3429) AgeI (3562) BclI * (3584) BsrGI (3611) SfiI (4037) PshAI (4201) EcoRI (4270) pZIP-mEF1-alpha-RFP-Puro 9114 bp
SgrDI  (9112)
1 site
C G T C G A C G G C A G C T G C
SspI  (8995)
1 site
A A T A T T T T A T A A
PvuI  (8561)
1 site
C G A T C G G C T A G C
FspI  (8413)
1 site
T G C G C A A C G C G T
AhdI  (8191)
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.
PmeI  (6933)
1 site
G T T T A A A C C A A A T T T G
PsiI  (6883)
1 site
T T A T A A A A T A T T
PstI  (6424)
1 site
C T G C A G G A C G T C
AscI  (5806)
1 site
G G C G C G C C C C G C G C G G
MluI  (5799)
1 site
A C G C G T T G C G C A
HpaI  (5558)
1 site
G T T A A C C A A T T G
BamHI  (5472)
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.
NotI  (5462)
1 site
G C G G C C G C C G C C G G C G
BsmBI  (5253)
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.
BstEII  (4995)
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.
RsrII  (4977)
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.
BsiWI  (4917)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PflFI  (4903)
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  (4903)
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  (4826)
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.
PmlI  (4599)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
NdeI  (276)
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.
SnaBI  (382)
1 site
T A C G T A A T G C A T
NheI  (405)
1 site
G C T A G C C G A T C G
BmtI  (409)
1 site
G C T A G C C G A T C G
NruI  (867)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
BspQI  (1136)
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  (1136)
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.
FseI  (1184)
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.
MfeI  (1223)
1 site
C A A T T G G T T A A C
BbvCI  (1458)
1 site
C C T C A G C G G A G T C G
KflI  (1968)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
BspDI  (2214)
1 site
A T C G A T T A G C T A
ClaI  (2214)
1 site
A T C G A T T A G C T A
BlpI  (2344)
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.
BstXI  (3003)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
XcmI  (3429)
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.
AgeI  (3562)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BclI  (3584)
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.
BsrGI  (3611)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SfiI  (4037)
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.
PshAI  (4201)
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.
EcoRI  (4270)
1 site
G A A T T C C T T A A G
mEF-1α promoter
2219 .. 3562  =  1344 bp
strong constitutive promoter for mouse elongation
factor EF-1α
mEF-1α promoter
2219 .. 3562  =  1344 bp
strong constitutive promoter for mouse elongation
factor EF-1α
AmpR
8118 .. 8978  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   8118 .. 8909  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8118 .. 8978  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8910 .. 8978  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8118 .. 8978  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
TurboRFP
3574 .. 4269  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
TurboRFP
3574 .. 4269  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
PuroR
4858 .. 5460  =  603 bp
200 amino acids  =  21.6 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4858 .. 5460  =  603 bp
200 amino acids  =  21.6 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
5819 .. 6407  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5819 .. 6407  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7362 .. 7947  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7362 .. 7947  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
4283 .. 4855  =  573 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
4283 .. 4855  =  573 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
6944 .. 7273  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
6944 .. 7273  =  330 bp
SV40 enhancer and early promoter
5' LTR (truncated)
411 .. 668  =  258 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
411 .. 668  =  258 bp
truncated 5' long terminal repeat (LTR) from HIV-1
RRE
1337 .. 1570  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1337 .. 1570  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
3' LTR (ΔU3)
6615 .. 6848  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
6615 .. 6848  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' miR-30a
5672 .. 5798  =  127 bp
sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)
3' miR-30a
5672 .. 5798  =  127 bp
sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)
HIV-1 Ψ
715 .. 840  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
715 .. 840  =  126 bp
packaging signal of human immunodeficiency virus
type 1
5' miR-30a
5481 .. 5602  =  122 bp
sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)
5' miR-30a
5481 .. 5602  =  122 bp
sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)
cPPT/CTS
2062 .. 2177  =  116 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
cPPT/CTS
2062 .. 2177  =  116 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
AmpR promoter
8979 .. 9083  =  105 bp
AmpR promoter
8979 .. 9083  =  105 bp
shRNA
5603 .. 5671  =  69 bp
   Segment 1:  
   5603 .. 5629  =  27 bp
short hairpin RNA
shRNA
5603 .. 5671  =  69 bp
   Segment 2:  miR-30a loop  
   5630 .. 5644  =  15 bp
short hairpin RNA
shRNA
5603 .. 5671  =  69 bp
   Segment 3:  
   5645 .. 5671  =  27 bp
short hairpin RNA
shRNA
5603 .. 5671  =  69 bp
3 segments
short hairpin RNA
mEF-1α intron
2583 .. 3530  =  948 bp
intron upstream of the start codon of mouse EF-1α
mEF-1α intron
2583 .. 3530  =  948 bp
intron upstream of the start codon of mouse EF-1α
SV40 ori
7124 .. 7259  =  136 bp
SV40 origin of replication
SV40 ori
7124 .. 7259  =  136 bp
SV40 origin of replication
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter