pRetroQ-AcGFP1-C1

Self-inactivating retroviral vector for expressing a protein fused to the C-terminus of AcGFP1.

Sequence Author: Clontech (TaKaRa)

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SspI (7269) XmnI (7064) ScaI (6945) FspI (6687) AflIII - PciI (5572) BspQI - SapI (5456) AvrII (5325) SfiI (5278) EcoRV (4201) CMV enhancer AscI (542) PshAI (676) NheI (2172) BmtI (2176) PflMI (2892) BglII (2920) PaeR7I - XhoI (2924) BstBI (2938) EcoRI (2940) SalI (2950) AccI (2951) PspOMI (2964) ApaI (2968) BamHI (2971) XbaI * (2983) stop codons BclI * (2993) BlpI (3282) BsmI (3440) BfuAI - BspMI (3518) BsiWI (3585) RsrII (3645) pRetroQ-AcGFP1-C1 7614 bp
SspI  (7269)
1 site
A A T A T T T T A T A A
XmnI  (7064)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (6945)
1 site
A G T A C T T C A T G A
FspI  (6687)
1 site
T G C G C A A C G C G T
AflIII  (5572)
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  (5572)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (5456)
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  (5456)
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  (5325)
1 site
C C T A G G G G A T C C
SfiI  (5278)
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  (4201)
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.
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
PflMI  (2892)
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.
BglII  (2920)
1 site
A G A T C T T C T A G A
PaeR7I  (2924)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2924)
1 site
C T C G A G G A G C T C
BstBI  (2938)
1 site
T T C G A A A A G C T T
EcoRI  (2940)
1 site
G A A T T C C T T A A G
SalI  (2950)
1 site
G T C G A C C A G C T G
AccI  (2951)
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  (2964)
1 site
G G G C C C C C C G G G
ApaI  (2968)
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  (2971)
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  (2983)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (2993)
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.
BlpI  (3282)
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  (3440)
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.
BfuAI  (3518)
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  (3518)
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.
BsiWI  (3585)
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.
RsrII  (3645)
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.
AmpR
6392 .. 7252  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   6392 .. 7183  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6392 .. 7252  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   7184 .. 7252  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6392 .. 7252  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AcGFP1
2194 .. 2910  =  717 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
mammalian codon-optimized
AcGFP1
2194 .. 2910  =  717 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
mammalian codon-optimized
PuroR
3529 .. 4128  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3529 .. 4128  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
5633 .. 6221  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5633 .. 6221  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
3009 .. 3508  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
3009 .. 3508  =  500 bp
mouse phosphoglycerate kinase 1 promoter
3' LTR (ΔU3)
4313 .. 4745  =  433 bp
self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus
3' LTR (ΔU3)
4313 .. 4745  =  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
5011 .. 5340  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
5011 .. 5340  =  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
7253 .. 7357  =  105 bp
AmpR promoter
7253 .. 7357  =  105 bp
MCS
2920 .. 2976  =  57 bp
multiple cloning site
MCS
2920 .. 2976  =  57 bp
multiple cloning site
stop codons
2985 .. 2995  =  11 bp
stop codons in all three reading frames
stop codons
2985 .. 2995  =  11 bp
stop codons in all three reading frames
SV40 ori
5191 .. 5326  =  136 bp
SV40 origin of replication
SV40 ori
5191 .. 5326  =  136 bp
SV40 origin of replication
ORF:  2194 .. 2991  =  798 bp
ORF:  265 amino acids  =  29.3 kDa
ORF:  3289 .. 4128  =  840 bp
ORF:  279 amino acids  =  30.1 kDa
ORF:  6522 .. 6788  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  838 .. 1098  =  261 bp
ORF:  86 amino acids  =  9.5 kDa
ORF:  3454 .. 4179  =  726 bp
ORF:  241 amino acids  =  24.8 kDa
ORF:  4569 .. 4859  =  291 bp
ORF:  96 amino acids  =  10.5 kDa
ORF:  6392 .. 7252  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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