LOCUS       sequence-411068-        3607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fluorescent protein GFPmut2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3607)
  AUTHORS   Tchagang CF, Mah TF, Campbell-Valois FX
  TITLE     Anaerobic fluorescent reporters for live imaging of Pseudomonas 
            aeruginosa.
  JOURNAL   Front Microbiol. 2023 Oct 20;14:1245755. doi: 
            10.3389/fmicb.2023.1245755. eCollection 2023.
  PUBMED    37928662
REFERENCE   2  (bases 1 to 3607)
  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: "Front 
            Microbiol."; date: "2023-10-20"; volume: "14:1245755. doi"; pages: "
            10.3389/fmicb.2023.1245755. eCollection 2023"
COMMENT     Sequence Label: pCFT9.2
FEATURES             Location/Qualifiers
     source          1..3607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..772
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             773..1435
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     1436..1452
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1436..1452
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1563..1581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1593..1609
                     /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)."
     CDS             1674..2387
                     /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(1704..1732)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2327..2347
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2533..2550)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2533..2549)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2549..2620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2621..2689,2690..3481)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2839..2858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        3482..3500
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(join(3589..3607,1..2))
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3589..3607)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg
       61 gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt
      121 cgaccacata cccaccggct ccaactgcgc ggcctgtggc cttgccccat caattttttt
      181 aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg ccggttggac
      241 accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg gccttgacgc
      301 gcctggaacg acccaagcct atgcgagtgg gggcagtcga agggcgaagc ccgcccgcct
      361 gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc caccttgggc
      421 aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt tgccggaggg
      481 ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg gcaccaaaga
      541 actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa aggggggtac
      601 gcaacagctc attgcggcac cccccgcaat agctcattgc gtaggttaaa gaaaatctgt
      661 aattgactgc cacttttacg caacgcataa ttgttgtcgc gctgccgaaa agttgcagct
      721 gattgcgcat ggtgccgcaa ccgtgcggca cccctaccgc atggagataa gcatggccac
      781 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
      841 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
      901 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
      961 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     1021 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     1081 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     1141 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     1201 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     1261 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     1321 ggaggaatgg gaacgacgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     1381 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcagga
     1441 aacagctatg accatgcccg ggtaagtgac tagcggaaac acagaaaaaa gcccgcacct
     1501 gacagtgcgg gctttttttt tcgaccaaag gctccgaaga gccgagtgtc ttcggtgaat
     1561 tgtaatacga ctcactatag ggcgaattga cattgtgagc ggataacaat ataatgtgtg
     1621 gaattcgagc tcggtacccg gggatcctct agatttaaga aggagatata catatgagta
     1681 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     1741 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     1801 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
     1861 ctttcgcgta tggtcttcaa tgctttgcga gatacccaga tcatatgaaa cagcatgact
     1921 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
     1981 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
     2041 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat
     2101 acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag
     2161 ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
     2221 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca
     2281 cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     2341 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaataa atgtccagac
     2401 ctgcagcaat ggccgacacg gtctctgaga gaagacacgt cgctcttcga cgtaagtaac
     2461 tgactcggta ccaaattcca gaaaagaggc ctcccgaaag gggggccttt tttcgttttg
     2521 gtcctaagct taactggccg tcgttttaca ttcaaatatg tatccgctca tgagacaata
     2581 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2641 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2701 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2761 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2821 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2881 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2941 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3001 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3061 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3121 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3181 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3241 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3301 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3361 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3421 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3481 ataatacgac tcactatagg gtaactaatt gaggcgcgcc taccaccgtc aaaaaaaacg
     3541 gcgcttttta gcgccgtttt tatttttcaa ccttcctagg taagtgaccc tttagtgagg
     3601 gttaatt
//
