LOCUS       sequence-392605-        6209 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent labelling of bacterial cells with tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6209)
  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 6209)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-tdTomato
FEATURES             Location/Qualifiers
     source          1..6209
                     /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)."
     mobile_element  3726..3891
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            4050..4158
                     /note="incP origin of transfer"
     rep_origin      complement(4390..4978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4470..4489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5149..5940,5941..6009))
                     /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     5772..5791
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6010..6114)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6182..6200
                     /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 atgccgtaca
     2041 ggaacaggtg gtggcggcct tccgagcgtt cgtactgttc cacgatggtg tagtcttcgt
     2101 tgtgcgaggt gatgtccagc ttggtgtcca cgtagtagta gcccggcagc tgcaccggct
     2161 tcttggccat gtagatggtc ttgaattcca ccaggtagtg gccgccgtcc ttcagcttca
     2221 gggcctggtg gatttcgccc ttcagcacgc cgtcgcgcgg gtacaggcgt tcggtcgagg
     2281 cttcccagcc catggtcttc ttctgcatca ccgggccgtc cggcgggaag ttggtgccgc
     2341 gcatcttcac cttgtagatc agggtgccgt cctgcagcga cgagtcctgg gtcacggtca
     2401 ccaggccgcc gtcttcgaag ttcatcacgc gttcccactt gaagccttcc gggaacgaca
     2461 gcttcttgta gtccgggatg tcggccgggt gcttcacgta ggccttcgag ccgtacatga
     2521 actgcggcga caggatgtcc caggcgaacg gcagcgggcc gcccttggtc accttcagct
     2581 tggcggtctg ggtgccttcg tacgggcggc cttcgccttc gccttcgatt tcgaattcgt
     2641 ggccgttcat cgagccttcc atgcgcacct tgaagcgcat gaattccttg atcacggcca
     2701 tgttgttgtc ttccgacgag gcggtgcccg acgagcccga gccggtcgag ccggtgccgt
     2761 ggcccaggaa caggtggtgg cggccttccg agcgttcgta ctgttccacg atggtgtagt
     2821 cttcgttgtg cgaggtgatg tccagcttgg tgtccacgta gtagtagccc ggcagctgca
     2881 ccggcttctt ggccatgtag atggtcttga attccaccag gtagtggccg ccgtccttca
     2941 gcttcagggc ctggtggatt tcgcccttca gcacgccgtc gcgcgggtac aggcgttcgg
     3001 tcgaggcttc ccagcccatg gtcttcttct gcatcaccgg gccgtccggc gggaagttgg
     3061 tgccgcgcat cttcaccttg tagatcaggg tgccgtcctg cagcgacgag tcctgggtca
     3121 cggtcaccag gccgccgtct tcgaagttca tcacgcgttc ccacttgaag ccttccggga
     3181 acgacagctt cttgtagtcc gggatgtcgg ccgggtgctt cacgtaggcc ttcgagccgt
     3241 acatgaactg cggcgacagg atgtcccagg cgaacggcag cgggccgccc ttggtcacct
     3301 tcagcttggc ggtctgggtg ccttcgtacg ggcggccttc gccttcgcct tcgatttcga
     3361 attcgtggcc gttcatcgag ccttccatgc gcaccttgaa gcgcatgaat tccttgatca
     3421 cttcttcgcc cttcgacacc atgtagtcct cctgccaccg tttaattcgg atccacgggt
     3481 tcgctacctg cattacagcc tacagaccga gaaggcttat gtctactggg ccaaggcatt
     3541 tgtgttgtgg acggcccgca gccatggtgg gtttcgacat ccgcgcgaaa tggggcaagc
     3601 tgaagtcgag ggttttctga ccatgctcgc caccgagaag caagtggcgc cggccaccca
     3661 ccggcaggcg ctcaacgcgc tgttgttctt gtatcggcag gtacctcgcg aaggccttgc
     3721 aggccaacca gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat
     3781 tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt
     3841 aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat
     3901 ttttcttcct gttatgtttg ggcgctgcat taatgaatcg gccaacgcgc ggggagaggc
     3961 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg acccgctgcg ctcggtcgtt
     4021 cggctgcggc gagcggtatc agagcttatc ggccagcctc gcagagcagg attcccgttg
     4081 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     4141 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     4201 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     4261 atgaccccga agcagggtta tgcagcggaa agtatacctt aaggaatccc ctgataacgc
     4321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcgag
     4441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4561 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4801 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5161 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5221 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5281 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5341 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5401 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5461 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5521 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5581 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5641 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5701 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5761 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5821 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5881 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5941 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6001 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6061 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6121 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6181 aaataggcgt atcacgaggc cctttcgtc
//
