LOCUS       sequence-224640-        1172 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  fluorescent protein expression with an N-terminal His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1172)
  AUTHORS   Holzapfel HY, Stern AD, Bouhaddou M, Anglin CM, Putur D, Comer S, 
            Birtwistle MR
  TITLE     Fluorescence Multiplexing with Spectral Imaging and Combinatorics.
  JOURNAL   ACS Comb Sci. 2018 Nov 12;20(11):653-659. doi: 
            10.1021/acscombsci.8b00101. Epub 2018 Oct 30.
  PUBMED    30339749
REFERENCE   2  (bases 1 to 1172)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acscombsci.8b00101"; journalName: "ACS Comb Sci"; date: 
            "2018-11-12- 12"; volume: "20"; issue: "11"; pages: "653-659"
COMMENT     Sequence Label: pQLinkHD_EBFP2-ECFP
FEATURES             Location/Qualifiers
     source          1..1172
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     21..39
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(89..103,104..109,110..126,127..133)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    109..125
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    141..157
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             176..187
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             200..220
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    233..257
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(260..262,263..265,266..976)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(305..326)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 tnntgacntt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgagc
       61 ttaattaaca acaccatttg tcgagaaatc ataaaaaatt tatttgcttt gtgagcggat
      121 aacaattata atagattcaa ttgtgagcgg ataacaattt cacacagaat tcattaaaga
      181 ggagaaatta actatgaaac atcaccatca ccatcaccat ggtgctgcag gtacaagttt
      241 gtacaaaaaa gcaggcttca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      301 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtga ggggcgaggg
      361 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      421 gcccgtgccc tggcccaccc tcgtgaccac cctgagccac ggcgtgcagt gcttcgcccg
      481 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      541 ccaggagcgc accatcttct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa
      601 gttcgagggc gacaccctag tgaaccgcat cgagctgaag ggcgtcgact tcaaggagga
      661 cggcaacatc ctggggcaca agctggagta caacttcaac agccacaaca tctatatcat
      721 ggccgtcaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acgtggagga
      781 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
      841 gctgctgccc gacagccact acctgagcac ccagtccgtg ctgagcaaag accccaacga
      901 gaagcgcgat cacatggtcc tgctggagtt ccgcaccgcc gccgggatca ctctcggcat
      961 ggacgagctg tacaagaccg gtgccggaan nngggggcct angaatggtg nncaanggcc
     1021 aagggcgatg cnacctacgg caagctgacc ctgaagttca tntgnnccac cggcaagctg
     1081 cccgtgccnt ggccnaccct cnggaccncc ctgaactggg ggcgtgcagt gcttcaccnn
     1141 nnacccnaac nnanggaann naccaanntt tt
//
