LOCUS       sequence-410862-        3272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fluorescent protein FAST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3272)
  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 3272)
  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: pCFT7.2
FEATURES             Location/Qualifiers
     source          1..3272
                     /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..2048
                     /codon_start=1
                     /product="small protein tag, derived from photoactive 
                     yellow protein, that allows in vivo fluorescent labeling by
                     addition of a cell-permeant fluorogen (Plamont et al., 
                     2016)"
                     /label=Y-FAST
                     /translation="MEHVAFGSEDIENTLAKMDDGQLDGLAFGAIQLDGDGNILQYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDSPEFYGKFKEGVASGNLNTMFEWMIPTSRGPTK
                     VKVHMKKALSGDSYWVFVKRV"
     primer_bind     complement(2198..2215)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2198..2214)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2214..2285
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2286..2354,2355..3146)
                     /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(2504..2523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        3147..3165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(join(3254..3272,1..2))
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3254..3272)
                     /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 ggcctgcggc 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 catatggaac
     1681 atgtggcgtt cgggtccgag gatatcgaga atacgctggc caagatggat gatggccagc
     1741 tggatggcct ggcgttcggc gccatccaac tcgacggcga cgggaatatc ctccagtata
     1801 atgcggcgga aggggatatc acgggccgcg atcccaaaca ggtgatcggc aagaatttct
     1861 tcaaggacgt cgcccccggg accgattcgc cggagttcta tggcaagttc aaggaagggg
     1921 tggcctcggg gaatctcaat acgatgttcg aatggatgat cccgacgtcg cgcggcccca
     1981 cgaaagtgaa ggtccatatg aaaaaggccc tgtccgggga ttcgtattgg gtcttcgtga
     2041 agcgcgtgta actgaatgtc cagacctgca gcaatggccg acacggtctc tgagagaaga
     2101 cacgtcgctc ttcgacgtaa gtaactgact cggtaccaaa ttccagaaaa gaggcctccc
     2161 gaaagggggg ccttttttcg ttttggtcct aagcttaact ggccgtcgtt ttacattcaa
     2221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2281 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2341 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2401 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2461 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2521 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2581 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2641 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2701 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2761 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2821 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2881 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2941 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3001 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3061 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3121 gtgcctcact gattaagcat tggtaataat acgactcact atagggtaac taattgaggc
     3181 gcgcctacca ccgtcaaaaa aaacggcgct ttttagcgcc gtttttattt ttcaaccttc
     3241 ctaggtaagt gaccctttag tgagggttaa tt
//
