BBa_K2022012
1
BBa_K2022012
INPNC-GFP, i.e. membrane-anchored GFP
2016-10-11T11:00:00Z
2016-10-12T03:35:40Z
To make this part, we combined BBa_K811005 (INPNC) with BBa_E0040 (GFP).
First, we digested BBa_K811005 with BamHI and PstI.
Then, we PCR amplified BBa_E0040 using Suffix-R2, and a forward oligonucleotides that replaced the prefix with a BamHI site (aggcggatccggtatgcgtaaaggagaagaacttttcactg). This PCR product was then also digested with BamHI and PstI.
These two products were then ligated together, and transformed into E. coli host (DH5-alpha derivative)
A commonly used reporter is Green Fluorescent Protein (GFP) (BBa_E0040). Here, we have fused it to the C-terminus of the Ice Nucleation Protein, N- and C-termini (BBa_K811005), which is a well-characterised membrane anchor. As such, this part is intended to function like GFP, but simply localised to the membrane of cells
false
false
_2489_
31054
31054
9
false
Pennsylvania (2012) designed their INPNC part with the BamHI site, to make it readily fusible to other proteins; we thank them for their contribution in making the assembly and design processes as simple as possible.
As such, the only design consideration was to use the oligonucleotide aggcggatccggtatgcgtaaaggagaagaacttttcactg to add a BamHI site upstream of the GFP open reading frame
false
Jessica Mazalo, Daniel Winter
annotation2493573
1
GFP
range2493573
1
961
1680
annotation2493572
1
INPNC
range2493572
1
1
957
annotation2493571
1
double stop
range2493571
1
1675
1680
annotation2493566
1
start
range2493566
1
1
3
BBa_K2022012_sequence
1
atgaccctggacaaagctctggttctgcgtacctgcgctaacaacatggctgaccactgcggtctgatctggccggcttctggtaccgttgaatctcgttactggcagtctacccgtcgtcacgaaaacggtctggttggtctgctgtggggtgctggtacctctgctttcctgtctgttcacgctgacgctcgttggatcgtttgcgaagttgctgttgctgacatcatctctctggaagaaccgggtatggttaaattcccgcgtgctgaagttgttcacgttggtgaccgtatctctgcttctcacttcatctctgctcgtcaggctgacccggcttctacctctacctctacctctacctctaccctgaccccgatgccgaccgctatcccgaccccgatgccggctgttgcttctgttaccctgccggttgctgaacaggctcgtcacgaagttttcgacgttgcttctgtttctgctgctgctgctccggttaacaccctgccggttaccaccccgcagaacctgcaaaccgctacctacggttctaccctgtctggtgacaaccactctcgtctgatcgctggttacggttctaacgaaaccgctggtaaccactctgacctgatcggtggtcacgactgcaccctgatggctggtgaccagtctcgtctgaccgctggtaaaaactctgttctgaccgctggtgctcgttctaaactgatcggttctgaaggttctaccctgtctgctggtgaagactctaccctgatcttccgtctgtgggacggtaaacgttaccgtcagctggttgctcgtaccggtgaaaacggtgttgaagctgacatcccgtactacgttaacgaagacgacgacatcgttgacaaaccggacgaagacgacgactggatcgaagttaaaggtggcggtagtggcggtgcaggaggcggatccggtatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z