Composite

Part:BBa_K1983018

Designed by: Vykintas Jauniškis   Group: iGEM16_Vilnius-Lithuania   (2016-10-13)
Revision as of 21:36, 21 October 2016 by Vykintas (Talk | contribs)


PheP under constitutive promoter, low 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, low 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/Low 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.


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]


[edit]
Categories
Parameters
None