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URL: http://www.addgene.org/240643

Proper Citation: RRID:Addgene_240643

Insert Name: AIPGENE865 promoter

Organism: Exaiptasia diaphana

Bacterial Resistance: Ampicillin

Vector Backbone Description: Backbone Marker:Tim Howes; Backbone Size:3854; Vector Backbone:pT2HE; Vector Types:Gene expression and genomic integration in fish; Bacterial Resistance:Ampicillin

Comments: The whole intergenic region between AIPGENE865 (KXJ12416.1; Elongation Factor 1alpha) and AIPGENE899 was cloned as promoter fragment. The original gene annotation for AIPGENE865 is incorrect, the automatic NCBI annotation is correct in regards of exons, introns and translation start site. We found that Aiptasia strain CC7 was actually heterozygous for that locus: one allele was mostly in agreement with the genome sources but the other one had a couple of SNPs and a large, 749-bp insert in the intergenic region. The longer version with insert was used as promoter for this plasmid. The Aiptasia MEK2 locus was found to be erroneously split into AIPGENES3280 & 23794 on two different scaffolds due to an assembly gap in the Baumgarten et al., 2015 genome. Correct CDS was identified from sequencing based on cDNA of strain CC7. AmNeonGreen is a version of mNeonGreen codon-optimized for Aiptasia (based on the transcriptome of Lehnert et al., 2014) with the identical protein sequence. Baumgarten S, Simakov O, Esherick LY, Liew YJ, Lehnert EM, Michell CT, Li Y, Hambleton EA, Guse A, Oates ME, et al. 2015. The genome of Aiptasia, a sea anemone model for coral symbiosis. Proc Natl Acad Sci. 112(38):11893–11898. doi:10.1073/pnas.1513318112. Lehnert EM, Mouchka ME, Burriesci MS, Gallo ND, Schwarz JA, Pringle JR. 2014. Extensive differences in gene expression between symbiotic and aposymbiotic cnidarians. G3 Bethesda Md. 4(2):277–295. doi:10.1534/g3.113.009084.

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