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Plasmid Files

pRetroQ-DsRed-Monomer-N1

Self-inactivating retroviral vector for expressing a protein fused to the N-terminus of DsRed-Monomer.

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pRetroQ-DsRed-Monomer-N1 Sequence and MappRetroQ-DsRed-Monomer-N1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SspI (7226) XmnI (7021) ScaI (6902) AflIII - PciI (5529) BspQI - SapI (5413) AvrII (5282) SfiI (5235) EcoRV (4158) RsrII (3602) CMV enhancer AscI (542) PshAI (676) CMV enhancer NheI (2172) BmtI (2176) PaeR7I - XhoI (2194) BstBI (2208) EcoRI (2210) SalI (2220) AccI (2221) PspOMI (2234) ApaI (2238) BamHI (2241) FspAI (2308) SbfI (2600) NgoMIV (2680) NaeI (2682) PflMI (2841) AleI (2887) NotI (2940) XbaI (2950) BlpI (3239) BsmI (3397) BsiWI (3542) pRetroQ-DsRed-Monomer-N1 7571 bp
SspI  (7226)
1 site
A A T A T T T T A T A A
XmnI  (7021)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (6902)
1 site
A G T A C T T C A T G A
AflIII  (5529)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (5529)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (5413)
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  (5413)
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.
AvrII  (5282)
1 site
C C T A G G G G A T C C
SfiI  (5235)
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.
EcoRV  (4158)
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.
RsrII  (3602)
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.
AscI  (542)
1 site
G G C G C G C C C C G C G C G G
PshAI  (676)
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.
NheI  (2172)
1 site
G C T A G C C G A T C G
BmtI  (2176)
1 site
G C T A G C C G A T C G
PaeR7I  (2194)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2194)
1 site
C T C G A G G A G C T C
BstBI  (2208)
1 site
T T C G A A A A G C T T
EcoRI  (2210)
1 site
G A A T T C C T T A A G
SalI  (2220)
1 site
G T C G A C C A G C T G
AccI  (2221)
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.
PspOMI  (2234)
1 site
G G G C C C C C C G G G
ApaI  (2238)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BamHI  (2241)
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.
FspAI  (2308)
1 site
R T G C G C A Y Y A C G C G T R
SbfI  (2600)
1 site
C C T G C A G G G G A C G T C C
NgoMIV  (2680)
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.
NaeI  (2682)
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.
PflMI  (2841)
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.
AleI  (2887)
1 site
C A C N N N N G T G G T G N N N N C A C
NotI  (2940)
1 site
G C G G C C G C C G C C G G C G
XbaI  (2950)
1 site
T C T A G A A G A T C T
BlpI  (3239)
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.
BsmI  (3397)
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.
BsiWI  (3542)
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.
AmpR
6349 .. 7209  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   6349 .. 7140  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6349 .. 7209  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   7141 .. 7209  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6349 .. 7209  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
DsRed-Monomer
2260 .. 2937  =  678 bp
225 amino acids  =  25.4 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
DsRed-Monomer
2260 .. 2937  =  678 bp
225 amino acids  =  25.4 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
PuroR
3486 .. 4085  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3486 .. 4085  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
5590 .. 6178  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5590 .. 6178  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
PGK promoter
2966 .. 3465  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
2966 .. 3465  =  500 bp
mouse phosphoglycerate kinase 1 promoter
3' LTR (ΔU3)
4270 .. 4702  =  433 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
3' LTR (ΔU3)
4270 .. 4702  =  433 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
gag (truncated)
1151 .. 1567  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
1151 .. 1567  =  417 bp
truncated MMLV gag gene lacking the start codon
SV40 promoter
4968 .. 5297  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4968 .. 5297  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
5 .. 308  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
5 .. 308  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1642 .. 1945  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1642 .. 1945  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
309 .. 512  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
309 .. 512  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
1946 .. 2149  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
1946 .. 2149  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
MMLV Ψ
751 .. 950  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
751 .. 950  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
5' LTR (truncated)
513 .. 688  =  176 bp
truncated long terminal repeat from Moloney murine
sarcoma virus
5' LTR (truncated)
513 .. 688  =  176 bp
truncated long terminal repeat from Moloney murine
sarcoma virus
AmpR promoter
7210 .. 7314  =  105 bp
AmpR promoter
7210 .. 7314  =  105 bp
MCS
2172 .. 2246  =  75 bp
multiple cloning site
MCS
2172 .. 2246  =  75 bp
multiple cloning site
SV40 ori
5148 .. 5283  =  136 bp
SV40 origin of replication
SV40 ori
5148 .. 5283  =  136 bp
SV40 origin of replication
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