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Plasmid Files

pTandem™-1

Mammalian vector for the co-expression of two genes from a bicistronic mRNA using the CMV promoter and an IRES.

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pTandem-1 Sequence and MappTandem-1.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 Bpu10I (3785) SspI (3752) ScaI (3428) PvuI (3318) FspI (3170) NmeAIII (3096) BpmI (3018) AhdI (2948) PspFI (2521) BseYI (2517) DrdI (2321) BstAPI (2041) BamHI (3806) BtgZI (329) SnaBI (335) BsgI (669) SacII (726) RsrII (901) NcoI (991) XcmI (1017) TspMI - XmaI (1093) SmaI (1095) PshAI (1135) MfeI (1140) EcoRV (1147) AscI - BssHII (1152) NsiI (1163) PacI (1168) SwaI (1174) BsiWI (1183) SfiI (1186) BstEII (1192) AgeI (1196) NotI (1204) PvuII (1214) AccI (1218) BstZ17I (1219) PaeR7I - XhoI (1266) Bsu36I (1308) SpeI (1314) AvrII (1397) HindIII (1470) AarI (1585) Acc65I (1687) KpnI (1691) BmgBI (1788) PmeI (1809) NheI (1814) BmtI (1818) AfeI (1824) BstXI (1826) HpaI (1832) BlpI (1837) BspDI * - ClaI * (1844) BspHI (1880) pTandem™-1 3813 bp
Bpu10I  (3785)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
SspI  (3752)
1 site
A A T A T T T T A T A A
ScaI  (3428)
1 site
A G T A C T T C A T G A
PvuI  (3318)
1 site
C G A T C G G C T A G C
FspI  (3170)
1 site
T G C G C A A C G C G T
NmeAIII  (3096)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BpmI  (3018)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AhdI  (2948)
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.
PspFI  (2521)
1 site
C C C A G C G G G T C G
BseYI  (2517)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
DrdI  (2321)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BstAPI  (2041)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BamHI  (3806)
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.
BtgZI  (329)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
SnaBI  (335)
1 site
T A C G T A A T G C A T
BsgI  (669)
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).
SacII  (726)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
RsrII  (901)
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.
NcoI  (991)
1 site
C C A T G G G G T A C C
XcmI  (1017)
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.
TspMI  (1093)
1 site
C C C G G G G G G C C C
XmaI  (1093)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (1095)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PshAI  (1135)
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.
MfeI  (1140)
1 site
C A A T T G G T T A A C
EcoRV  (1147)
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  (1152)
1 site
G G C G C G C C C C G C G C G G
BssHII  (1152)
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.
NsiI  (1163)
1 site
A T G C A T T A C G T A
PacI  (1168)
1 site
T T A A T T A A A A T T A A T T
SwaI  (1174)
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.
BsiWI  (1183)
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.
SfiI  (1186)
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.
BstEII  (1192)
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.
AgeI  (1196)
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.
NotI  (1204)
1 site
G C G G C C G C C G C C G G C G
PvuII  (1214)
1 site
C A G C T G G T C G A C
AccI  (1218)
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.
BstZ17I  (1219)
1 site
G T A T A C C A T A T G
PaeR7I  (1266)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1266)
1 site
C T C G A G G A G C T C
Bsu36I  (1308)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
SpeI  (1314)
1 site
A C T A G T T G A T C A
AvrII  (1397)
1 site
C C T A G G G G A T C C
HindIII  (1470)
1 site
A A G C T T T T C G A A
AarI  (1585)
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 AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
Acc65I  (1687)
1 site
G G T A C C C C A T G G
KpnI  (1691)
1 site
G G T A C C C C A T G G
BmgBI  (1788)
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.
PmeI  (1809)
1 site
G T T T A A A C C A A A T T T G
NheI  (1814)
1 site
G C T A G C C G A T C G
BmtI  (1818)
1 site
G C T A G C C G A T C G
AfeI  (1824)
1 site
A G C G C T T C G C G A
BstXI  (1826)
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.
HpaI  (1832)
1 site
G T T A A C C A A T T G
BlpI  (1837)
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.
BspDI  (1844)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1844)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BspHI  (1880)
1 site
T C A T G A A G T A C T
bla(M)
2875 .. 3663  =  789 bp
262 amino acids  =  28.8 kDa
Product: β-lactamase lacking the signal sequence
allows cytosolic expression of β-lactamase
bla(M)
2875 .. 3663  =  789 bp
262 amino acids  =  28.8 kDa
Product: β-lactamase lacking the signal sequence
allows cytosolic expression of β-lactamase
ori
2275 .. 2862  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2275 .. 2862  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
1321 .. 1805  =  485 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
1321 .. 1805  =  485 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
CMV enhancer
56 .. 359  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
56 .. 359  =  304 bp
human cytomegalovirus immediate early enhancer
MCS 1
991 .. 1271  =  281 bp
multiple cloning site 1
MCS 1
991 .. 1271  =  281 bp
multiple cloning site 1
β-globin terminator
1846 .. 2055  =  210 bp
rabbit β-globin terminator
β-globin terminator
1846 .. 2055  =  210 bp
rabbit β-globin terminator
CMV promoter
361 .. 560  =  200 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
361 .. 560  =  200 bp
human cytomegalovirus (CMV) immediate early
promoter
intron
679 .. 842  =  164 bp
intron
679 .. 842  =  164 bp
AmpR promoter
3736 .. 3807  =  72 bp
AmpR promoter
3736 .. 3807  =  72 bp
8xHis
1272 .. 1295  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
1272 .. 1295  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
T7 promoter
911 .. 929  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
911 .. 929  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
977 .. 982  =  6 bp
ribosome binding site
RBS
977 .. 982  =  6 bp
ribosome binding site
β-globin poly(A) signal
1913 .. 1968  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
1913 .. 1968  =  56 bp
rabbit β-globin polyadenylation signal
S-Tag
1026 .. 1070  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
S-Tag
1026 .. 1070  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
MCS 2
1806 .. 1848  =  43 bp
multiple cloning site 2
MCS 2
1806 .. 1848  =  43 bp
multiple cloning site 2
HSV tag
1233 .. 1265  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
1233 .. 1265  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
6xHis
999 .. 1016  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
999 .. 1016  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
thrombin site
1086 .. 1103  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
1086 .. 1103  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
enterokinase site
1122 .. 1136  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
1122 .. 1136  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
ATG
993 .. 995  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
993 .. 995  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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