LOCUS       sequence-307213-        1038 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Constitutive expression (sigma 70 promoter) of GCaMP6f tethered to 
            pHuji fluorophores in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1038)
  AUTHORS   Bruni GN, Kralj JM
  TITLE     Membrane voltage dysregulation driven by metabolic dysfunction 
            underlies bactericidal activity of aminoglycosides.
  JOURNAL   eLife. 2020 Aug 4;9. pii: 58706. doi: 10.7554/eLife.58706.
  PUBMED    32748785
REFERENCE   2  (bases 1 to 1038)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife."; 
            date: "2020-08-4"; volume: "9. pii: 58706. doi"; pages: " 
            10.7554/eLife.58706"
COMMENT     Sequence Label: pKL12-GCaMP6f-pHuji bb118
FEATURES             Location/Qualifiers
     source          1..1038
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(34..77)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    80..101
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        102..136
                     /label=BBa_J23118
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.56"
     CDS             176..193
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             197..229
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(233..241,242..256)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      263..272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             284..340
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     553..574
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(680..701)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ttnncctata aantaggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
       61 tcgctaagga tgatttctgg aattcgcggc cgcttctaga gttgacggct agctcagtcc
      121 taggtattgt gctagcatta actttaagaa ggagatatac atacccatgg gttctcatca
      181 tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta
      241 cgacgatgac gataaggatc tcgccaccat ggtcgactca tcacgtcgta agtggaataa
      301 gacaggtcac gcagtcagag ctataggtcg gctgagctca ctcgagaacg tctatatcaa
      361 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
      421 cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt
      481 gctgctgccc gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga
      541 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
      601 ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt
      661 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      721 cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      781 caccaccggc aagctgcccg tgccctnncc caccctcgtg accaccctga cctacggcgt
      841 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      901 gcccgannnt acatccagga gcgcaccatc ttnntcannc gacggnanta canacccgcg
      961 ccgagtgagt ngaggngana ccctnnnann catcgagctg nnnnatcnac ttcnnngnng
     1021 cannntnctg gggcnnna
//
