LOCUS       sequence-392611-        5489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent labelling of bacterial cells with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5489)
  AUTHORS   Mamat U, Hein M, Grella D, Taylor CS, Scholzen T, Alio I, Streit WR,
            Huedo P, Coves X, Conchillo-Sole O, Gomez AC, Gibert I, Yero D, 
            Schaible UE
  TITLE     Improved mini-Tn7 Delivery Plasmids for Fluorescent Labeling of 
            Stenotrophomonas maltophilia.
  JOURNAL   Appl Environ Microbiol. 2023 May 17:e0031723. doi: 
            10.1128/aem.00317-23.
  PUBMED    37195181
REFERENCE   2  (bases 1 to 5489)
  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: "Appl 
            Environ Microbiol. 2023 May 17:e0031723. doi: 10.1128/aem.00317-23."
COMMENT     Sequence Label: pUC18T-mini-Tn7T-Gm-Pc-mCherry
FEATURES             Location/Qualifiers
     source          1..5489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     584..600
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      826..912
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    958..1005
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1135..1668)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1857..1862,1863..1879,1880..1885))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1926..1973
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     2559..2577
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     mobile_element  3006..3171
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3330..3438
                     /note="incP origin of transfer"
     rep_origin      complement(3670..4258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3750..3769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4429..5220,5221..5289))
                     /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     5052..5071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5290..5394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5462..5480
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgacgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcag gtgtgggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc
      601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
      781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattgg ggatcttgaa
      961 gtacctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
     1021 ctgatgtgct taaaaactta ctcaatggaa ttctagatcg aattggccgc ggcgttgtga
     1081 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     1141 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
     1201 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
     1261 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
     1321 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacttggg
     1381 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
     1441 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
     1501 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
     1561 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
     1621 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
     1681 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     1741 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     1801 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
     1861 tacgaaccga acaggcttat gtcaattcga tccattgctg ttgacaaagg gaatcagggg
     1921 atcttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttcagagcgc
     1981 ttttgaagct aattcgatca tgcatgagct cttacttgta cagttcgtcc atgccgccgg
     2041 tcgagtggcg gccttcggcg cgttcgtact gttccacgat ggtgtagtct tcgttgtgcg
     2101 aggtgatgtc cagcttgatg ttcacgttgt aggcgcccgg cagctgcacc ggcttcttgg
     2161 ccttgtaggt ggtcttcact tcggcgtcgt agtggccgcc gtccttcagc ttcaggcgct
     2221 gcttgatttc gcccttcagg gcgccgtctt ccgggtacat gcgttccgac gaggcttccc
     2281 agcccatggt cttcttctgc atcaccgggc cgtccgacgg gaagttggtg ccgcgcagct
     2341 tcaccttgta gatgaattcg ccgtcctgca gcgacgagtc ctgggtcacg gtcaccacgc
     2401 cgccgtcttc gaagttcatc acgcgttccc acttgaagcc ttccgggaac gacagcttca
     2461 ggtagtccgg gatgtcggcc gggtgcttca cgtaggcctt cgagccgtac atgaactgcg
     2521 gcgacaggat gtcccaggcg aacggcagcg ggccgccctt ggtcaccttc agcttggcgg
     2581 tctgggtgcc ttcgtacggg cggccttcgc cttcgccttc gatttcgaat tcgtggccgt
     2641 tcaccgagcc ttccatgtgc accttgaagc gcatgaattc cttgatgatg gccatgttgt
     2701 cttcttcgcc cttcgacacc atgtagtcct cctgccaccg tttaattcgg atccacgggt
     2761 tcgctacctg cattacagcc tacagaccga gaaggcttat gtctactggg ccaaggcatt
     2821 tgtgttgtgg acggcccgca gccatggtgg gtttcgacat ccgcgcgaaa tggggcaagc
     2881 tgaagtcgag ggttttctga ccatgctcgc caccgagaag caagtggcgc cggccaccca
     2941 ccggcaggcg ctcaacgcgc tgttgttctt gtatcggcag gtacctcgcg aaggccttgc
     3001 aggccaacca gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat
     3061 tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt
     3121 aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat
     3181 ttttcttcct gttatgtttg ggcgctgcat taatgaatcg gccaacgcgc ggggagaggc
     3241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg acccgctgcg ctcggtcgtt
     3301 cggctgcggc gagcggtatc agagcttatc ggccagcctc gcagagcagg attcccgttg
     3361 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     3421 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     3481 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     3541 atgaccccga agcagggtta tgcagcggaa agtatacctt aaggaatccc ctgataacgc
     3601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcgag
     3721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4081 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4261 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4321 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4381 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4441 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4501 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4561 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4621 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4681 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4741 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4801 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4861 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4921 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4981 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5041 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5101 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5161 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5221 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5281 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5341 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5401 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5461 aaataggcgt atcacgaggc cctttcgtc
//
