Difference between revisions of "Part:BBa K1983017"

Latest revision as of 21:39, 21 October 2016

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 98
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K1983017 short</partinfo>
-BBa_K1983017
+<h2>Overview</h2>
+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.
+<h2>Experiments and Results</h2>
+<h3>Cloning</h3>
+This part was made by modifying ([https://parts.igem.org/Part:BBa_K1983014 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 ([https://parts.igem.org/Part:BBa_K1983014 BBa_K1983014]), which was cut by EcoRI and XbaI to delete the previous RBS. The site on ([https://parts.igem.org/Part:BBa_K1983014 BBa_K1983014]) for XbaI was designed intentionally for editing purposes to obtain this biobrick part.
+<h2>References</h2>
+<b>1.</b> 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.
 <!-- Add more about the biology of this part here