LOCUS       sequence-110014-         875 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Broad host range plasmid for constitutive expression of the eGFP. 
            Developed for imaging Pseudomonas aeruginosa in biofilms..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 875)
  AUTHORS   Nivens DE, Ohman DE, Williams J, Franklin MJ
  TITLE     Role of alginate and its O acetylation in formation of Pseudomonas 
            aeruginosa microcolonies and biofilms.
  JOURNAL   J Bacteriol. 2001 Feb;183(3):1047-57.
  PUBMED    11208804
REFERENCE   2  (bases 1 to 875)
  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: "J 
            Bacteriol."; date: "2001-02"; volume: "183(3)"; pages: "1047-57"
COMMENT     Sequence Label: pMF230
FEATURES             Location/Qualifiers
     source          1..875
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             36..749
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(66..94)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     689..709
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(789..806)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 atggctgcag tctagagtcg acaggagaag aaaaaatgag taaaggagaa gaacttttca
       61 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      121 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
      181 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcgcg tatggtcttc
      241 aatgctttgc gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc
      301 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
      361 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
      421 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aactcacaca
      481 atgtatacat catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac
      541 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
      601 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
      661 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
      721 ttacacatgg catggatgaa ctatacaaat aagcttggct gttttggcgg atgagagaag
      781 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
      841 cctggcggca gtagcgcggt ggtcccacct gaccc
//
