LOCUS       sequence-120478-         826 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of Venus-LUMP fusion with thrombin cleavage 
            site, a FRET-based thrombin sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 826)
  AUTHORS   Hoepker AC, Wang A, Le Marois A, Suhling K, Yan Y, Marriott G
  TITLE     Genetically encoded sensors of protein hydrodynamics and molecular 
            proximity.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Apr 30. pii: 201424021.
  PUBMED    25931526
REFERENCE   2  (bases 1 to 826)
  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: "Proc Natl 
            Acad Sci U S A. 2015 Apr 30. pii: 201424021."
COMMENT     Sequence Label: 274_pSKB3-Venus-LUMP
FEATURES             Location/Qualifiers
     source          1..826
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             48..65
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             87..107
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(111..113,114..116,117..824)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(156..177)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(417..436)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     764..785
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc
       61 atcacgatta cgatatccca acgaccgaaa acctttactt ccagggccat atggtgagca
      121 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      181 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      241 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      301 ccttcggcta cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact
      361 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      421 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      481 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      541 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
      601 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
      661 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
      721 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      781 tcgtgaccgc cgccgggatc actctcggca tggacgagct atacaa
//
