pTandem-1

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

Sequence Author: MilliporeSigma (Novagen)

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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) BspHI (1880) BamHI (3806) BtgZI (329) SnaBI (335) BsgI (669) SacII (726) RsrII (901) RBS NcoI (991) ATG 6xHis XcmI (1017) TspMI - XmaI (1093) SmaI (1095) enterokinase site 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) PaqCI (1585) Acc65I (1687) KpnI (1691) BmgBI (1788) PmeI (1809) NheI (1814) BmtI (1818) AfeI (1824) BstXI (1826) HpaI (1832) BlpI (1837) BspDI * - ClaI * (1844) pTandemâ„¢-1 3813 bp
Bpu10I  (3785)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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