LOCUS       sequence-224642-        1211 bp    DNA     linear   UNA 15-DEC-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 1211)
  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 1211)
  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"; 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-mTFP1
FEATURES             Location/Qualifiers
     source          1..1211
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     22..40
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(90..104,105..110,111..127,128..134)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    110..126
                     /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    142..158
                     /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             177..188
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             201..221
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    234..258
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(261..263,264..266,267..977)
                     /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(306..327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 atnntgacnt tnacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag
       61 cttaattaac aacaccattt gtcgagaaat cataaaaaat ttatttgctt tgtgagcgga
      121 taacaattat aatagattca attgtgagcg gataacaatt tcacacagaa ttcattaaag
      181 aggagaaatt aactatgaaa catcaccatc accatcacca tggtgctgca ggtacaagtt
      241 tgtacaaaaa agcaggcttc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      301 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg aggggcgagg
      361 gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      421 tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca cggcgtgcag tgcttcgccc
      481 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      541 tccaggagcg caccatcttc ttcaaggacg acggcaccta caagacccgc gccgaggtga
      601 agttcgaggg cgacacccta gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg
      661 acggcaacat cctggggcac aagctggagt acaacttcaa cagccacaac atctatatca
      721 tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacgtggagg
      781 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      841 tgctgctgcc cgacagccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg
      901 agaagcgcga tcacatggtc ctgctggagt tccgcaccgc cgccgggatc actctcggca
      961 tggacgagct gtacaagacc ggtgcccgga agtgggggcc taggaatggt gagcaagggc
     1021 gaggggaccn caatgggcgt aatcaagccc gacatgaaga tcaagctgaa nnatggaggg
     1081 caacgtgaat ggccacgcct tcnngatnna gggcaaggnc aagggcaagc cctacgacgg
     1141 cnccaanncn ttcaacnnng gaggnnaagg aggnnncccn nnnccttnnn ctnnnaatnn
     1201 ngaacccnng t
//
