LOCUS       sequence-307207-        1015 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Constitutive expression (sigma 70 promoter) of GCaMP6f tethered to 
            mScarlet fluorophores in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1015)
  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 1015)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pKL09-GCaMP6f-mScarlet bb118
FEATURES             Location/Qualifiers
     source          1..1015
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(27..70)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    73..94
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        95..129
                     /label=BBa_J23118
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.56"
     CDS             169..186
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             190..222
                     /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(226..234,235..249)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      256..265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             277..333
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     546..567
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(673..694)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 atnaantagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
       61 ggatgatttc tggaattcgc ggccgcttct agagttgacg gctagctcag tcctaggtat
      121 tgtgctagca ttaactttaa gaaggagata tacataccca tgggttctca tcatcatcat
      181 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      241 gacgataagg atctcgccac catggtcgac tcatcacgtc gtaagtggaa taagacaggt
      301 cacgcagtca gagctatagg tcggctgagc tcactcgaga acgtctatat caaggccgac
      361 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcggc
      421 gtgcagctcg cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      481 cccgacaacc actacctgag cgtgcagtcc aaactttcga aagaccccaa cgagnagcgc
      541 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      601 ctgtacaagg gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg
      661 gtggtgcccn tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      721 ggcgagggtg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      781 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      841 ttcagcngct accccgacnn nntgaagcag cacgacttct tcagtccgnc atgcccgaan
      901 gctacntcca gnncgccacc atcttcntca aggacnangg cnactacana nngcnnnagg
      961 tganttcgnn ggnnnnnnnc ctgnngaacc gcatcganct gaaggnatcg acttc
//
