I designed a plasmid to express one specific gene and I inserted the T7 terminator after stop codon of gene sequence… and when I checked it was reversed. I am wondering if it will work or not, and does it affect the protein expression if terminator is missing ?
The original sequence of t7 terminator =>
5’-ctagcataaccccttggggcctctaaacgggtcttgaggggttttttg-3’
What I did=>
5’-(gene sequence) caaaaaacccctcaagacccgtttagaggccccaaggggttatgctag-3’
I’d like to thank you for your advice and comment in advance Thank you!!
Normally terminators are directional, so it will most likely not work if oriented in reverse. However it may or may not be needed, depending upon your construct and application. The presence of the terminator is helpful to stabilize the plasmid so it's probably better to have it present, but in most cases it is not essential.
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