Experiment / E7AGYGAOOOther

Enhancer orientation tests in dual-luciferase reporters

Influence of genetic polymorphism on transcriptional enhancer activity in the malaria vector Anopheles coluzzii

Selected KLF, AP, and LRIM1 enhancer alleles were recloned in both arbitrary reporter orientations and assayed in A. coluzzii 4a3A cells. The source reports no detectable orientation effect for KLF/AP and a weak LRIM1 Fd05_#1 difference.

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Perturbation & assay details

Basal / Untreated; 24 h after reporter transfection

All conditions were run at n = 12. KLF and AP showed no detectable orientation effect; the paper reports p = 0.042 for the LRIM1 Fd05_#1 comparison but characterizes it as weak.

Episomal dual-luciferase orientation assay using pGL-Gateway-DSCP firefly reporter plus pRL-ubi-63E Renilla control; orientation A/B is a construct label, with normalized RLU output rather than barcode counts.

Processed data

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Column dictionary · 23 definitions
element_id
Enhancer locus tested.
proximal_gene
Nearest annotated gene used to name the locus.
gene_id
Gene identifier from the paper.
chromosome
Chromosome arm in PEST AgamP4 coordinates.
coordinate_start
1-based inclusive candidate interval start.
coordinate_end
1-based inclusive candidate interval end.
interval_length_bp
Inclusive candidate interval length.
allele_label
Allele label shown in Figure 5; AP Ng labels are preserved even where S1 uses an alternate clone header.
source_colony
Laboratory colony source of the enhancer haplotype.
colony_origin
Colony initiation population/year reported in Methods.
construct_type
Reporter construct design tested in this panel.
orientation
Arbitrary reporter-insert orientation A or B; these labels are not genomic plus/minus strands.
source_fasta_file
Supplementary File S1 FASTA filename retained in raw_data.
source_fasta_record
Exact S1 record used for the sequence column.
sequence_5to3
Ungapped nucleotide sequence reconstructed from the aligned FASTA record.
sequence_length_bp
Length of the ungapped sequence.
sequence_md5
MD5 checksum of the ungapped sequence.
relative_luciferase_activity_median_approx
Approximate Figure 5 median of DLX-normalized luciferase activity; digitized from the plotted median bar.
n_measurements
Figure 5 reports n = 12 wells for every orientation condition.
statistical_group
Compact-letter group printed above the Figure 5 distribution.
p_value_for_orientation_comparison
Only the paper-reported LRIM1 Fd05_#1 orientation comparison p-value is populated; blank means no pairwise p-value was stated.
source_figure
Publication figure supporting the activity value.
qc_pass
All source-tested orientations retained with n = 12; no paper-reported technical exclusions.

Quality control

Retained all 12 source-tested orientation conditions with n = 12. The reported weak p = 0.042 comparison is retained as a result, not used as an exclusion criterion.

Curation notes

Figure 5 uses AP labels Ng_#2 and Ng_#4, while Supplementary File S1 contains corresponding APT2_Ng1/Apt2_ECO1_Ng1 and Ng4 records; this source-label mismatch is preserved in allele_label/source_fasta_record rather than silently renamed.

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