Need eukaryote expression help - two promoters and just one terminator ok??

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Mega [Andreas Stuermer]

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Apr 27, 2014, 6:03:52 PM4/27/14
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Hi everyone! 

I want to do this: 


Promoter CMV ---- GFP ---- Promoter EF1A ---- Neomycin Resistance ---- Terminator SV40


Is that ok with eukaryotic cells? 

The CMV transcript will be Kozak -- GFP-- junk promoter ef1a --  junk neoR   --     (and properly terminated)

The EF1a transcript will be Kozak -- NeoResistance --(and properly terminated)


It was said here that weak transcription termination may lead to silencing or other unwanted effects.... 

Mega [Andreas Stuermer]

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Apr 27, 2014, 6:07:13 PM4/27/14
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Btw, 

There is no cheap way around it. Would add like 2000$ to change the vector, as it already contains the GFP.

I want to see GFP and NEO highly expressed
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