Part:BBa_C0053
c2 repressor from Salmonella phage P22 (+LVA)
The P22 c2 repressor protein coding sequence is a 720 base-pair sequence with the standard RBS-compatible BioBrick prefix and the standard BioBrick suffix sections on its ends. It binds to the P22 c2 regulatory sequence, BBa_R0053. The sequence contains a LVA tag for faster degradation.
Usage and Biology
P22 c2 is a member of the lamboid cI protein family.
The coding sequence has been modified from wild type c2 to contain an LVA tag,
and two TAA stop codons.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 166
Functional Parameters
direction | Forward |
function | Rep |
ligands | ? |
protein | C2 P22 |
swisspro | P69202 |
tag | LVA |
Functional Parameters: Austin_UTexas
Burden Imposed by this Part:
Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This part exhibited a significant burden. Users should be aware that BioBricks with a burden of >20-30% may be susceptible to mutating to become less functional or nonfunctional as an evolutionary consequence of this fitness cost. This risk increases as they used for more bacterial cell divisions or in larger cultures. Users should be especially careful when combining multiple burdensome parts, as plasmids with a total burden of >40% are expected to mutate so quickly that they become unclonable. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.
This functional parameter was added by the 2020 Austin_UTexas team.
Modification in Sequence : iGEM21_IISER-Tirupati_India
P22 c2 repressor DNA binding site.
Part Name | Part number | Sequence | Rel KD | KD (in M) |
---|---|---|---|---|
P22 binding site A | BBa_K3889080 | ATTTAAGATATCTTAAAT | 1 | 1.6 × 10-8 |
P22 binding site B | BBa_K3889081 | AATTAAGATATCTTAATT | 1.8 | 2.88 × 10-8 |
P22 binding site C | BBa_K3889082 | ATTTAAGAATTCTTAAAT | 2 | 3.2 × 10-8 |
P22 binding site D | BBa_K3889083 | AGTTAAGATATCTTAACT | 2.6 | 4.16 × 10-8 |
P22 binding site E | BBa_K3889084 | ATTAAAGATATCTTTAAT | 3.8 | 6.08 × 10-8 |
P22 binding site F | BBa_K3889085 | ACTTAAGATATCTTAAGT | 4.3 | 6.88 × 10-8 |
P22 binding site G | BBa_K3889086 | ATTCAAGATATCTTGAAT | 5 | 8.0 × 10-8 |
P22 binding site H | BBa_K3889087 | ATTGAAGATATCTTCAAT | 7.6 | 1.216 × 10-7 |
P22 binding site I | BBa_K3889088 | ATTTAAGAGCTCTTAAAT | 10 | 1.6 × 10-7 |
P22 binding site J | BBa_K3889089 | ATTTAAGACGTCTTAAAT | 10 | 1.6 × 10-7 |
P22 binding site K | BBa_K3889090 | ATTTACGATATCGTAAAT | 30 | 4.8 × 10-7 |
P22 binding site L | BBa_K3889091 | ATTTAAAATATTTTAAAT | 55 | 8.8 × 10-7 |
The alignment of these sequences are as follows:
Fig source: Geneious version 2021.2 created by Biomatters. Available from https://www.geneious.com
Usage and Biology
Different binding affinities of a repressor provides a variable system that can be used for different types of systems. As KD increases the dissociation of repressor from the DNA sequence also increases providing with lower repression. P22 repressor binds to this sequence as a dimer. This inhibits the enzymes from transcription the genes on whose promoter this operator site is present. Hence this could be fused with any promoter in order to form a repressible system as done in BBa_K3889102.
Another Contribution: Modification in sequence
BBa_K3889020 is a modified version specific to our project's assembly. The following modifications were made:
- Spa1 site Removed
- LVA tag Removed
- barcodes Removed
References
1. Watkins, D., Hsiao, C., Woods, K. K., Koudelka, G. B., & Williams, L. D. (2008). P22 c2 Repressor−Operator Complex: Mechanisms of Direct and Indirect Readout. Biochemistry, 47(8), 2325–2338. https://doi.org/10.1021/bi701826f
//cds/transcriptionalregulator/repressor
//function/regulation/transcriptional
direction | Forward |
function | Rep |
ligands | ? |
protein | C2 P22 |
swisspro | P69202 |
tag | LVA |