Part:BBa_K1983017
PheP under constitutive promoter, medium strength RBS and terminator
Overview
PheP (Phenylalanine-specific permease) is a natural phenylalanine membrane transporter of the bacterium Escherichia coli. PheP is a single integral membrane transporter which selectively transports L-phenylalanine and L-tyrosine as an antiporter using proton motive force. The activity of this transporter under natural expression is known to be 9 and 17,5 nmol/mgDW(cells) for L-phenylalanine and L-tyrosine respectively [1]. PheP gene is labeled with N-6XHis-Tag and is under a constitutive promoter, medium strength RBS and terminator.
Experiments and Results
Cloning
This part was made by modifying (BBa_K1983014). First, a fragment containing medium strength RBS was synthesized via PCR using RBS-FW/Med primers annealing to each other. The fragment was cut by EcoRI and SpeI and cloned into pSB1C3 vector containing (BBa_K1983014), which was cut by EcoRI and XbaI to delete the previous RBS. The site on (BBa_K1983014) for XbaI was designed intentionally for editing purposes to obtain this biobrick part.
References
1. Cosgriff, A. J., G. Brasier, et al. (2000). "A study of AroP-PheP chimeric proteins and identification of a residue involved in tryptophan transport." J Bacteriol 182(8): 2207-2217.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 98 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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