Difference between revisions of "Part:BBa K1983015"
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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 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]. | ||
| − | pheU gene from Escherichia coli encodes a tRNA transporting L-phenylalanine and is under an upstream constitutive promoter and a terminator found after the tRNA. | + | pheU gene from Escherichia coli encodes a tRNA transporting L-phenylalanine and is under an upstream constitutive promoter and a terminator found after the tRNA gene. |
| + | <h2>Experiments and results</h2> | ||
| + | |||
| + | <h1>Cloning</h1> | ||
| + | |||
| + | This part was created by fusing a fragment received from IDT that contains a part ([https://parts.igem.org/Part:BBa_K1983014 BBa_K1983014]) with a biobrick part ([https://parts.igem.org/Part:BBa_K1983011 BBa_K1983011]) inside a pSB1C3 backbone. The received sequences were amplified using Uni-FW/RV primers and digested with EcoRI and SpeI. The fragments were cloned into pSB1C3 vector containing ([https://parts.igem.org/Part:BBa_K1983011 BBa_K1983011]) digested with EcoRI and XbaI. | ||
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Revision as of 12:37, 22 October 2016
PheP and tRNA-Phe under constitutive promoters
Overview
This biobrick part contains two coding elements: (BBa_K1983014): PheP, which is labeled with N-6XHis-Tag and put under a constitutive promoter (BBa_K823005), high strength RBS 9BBa_J61132) and terminator (BBa_B0017). A basic biobrick part (BBa_K1983011): tRNA-Phe (pheU) under constitutive promoter and terminatorPheP (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].
pheU gene from Escherichia coli encodes a tRNA transporting L-phenylalanine and is under an upstream constitutive promoter and a terminator found after the tRNA gene.
Experiments and results
Cloning
This part was created by fusing a fragment received from IDT that contains a part (BBa_K1983014) with a biobrick part (BBa_K1983011) inside a pSB1C3 backbone. The received sequences were amplified using Uni-FW/RV primers and digested with EcoRI and SpeI. The fragments were cloned into pSB1C3 vector containing (BBa_K1983011) digested with EcoRI and XbaI.
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
Illegal NheI site found at 1669
Illegal NheI site found at 1692 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]