LOCUS       sequence-224651-         850 bp    DNA     linear   UNA 14-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 850)
  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 850)
  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"; 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_mVenus
FEATURES             Location/Qualifiers
     source          1..850
                     /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..725)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(306..327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(567..586)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 atnntgncat taacctataa 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 tccggcgagg
      361 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
      421 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc
      481 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      541 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      601 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
      661 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
      721 ccgccgacaa gcanaagaac ggcatcaagg ccaacttcaa gatccgccac aacatnnagg
      781 acggcggcgg gcagctcgcc gaacccctac caacaaaaan ccccccnccg gnnacggccc
      841 cngggcnnnc
//
