LOCUS       sequence-215252-        5778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini-Tn7 vector for insertion of DsRed-Express2 at bacterial attTn7 
            site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5778)
  AUTHORS   Wilton R, Ahrendt AJ, Shinde S, Sholto-Douglas DJ, Johnson JL, 
            Brennan MB, Kemner KM
  TITLE     A New Suite of Plasmid Vectors for Fluorescence-Based Imaging of 
            Root Colonizing Pseudomonads.
  JOURNAL   Front Plant Sci. 2018 Feb 1;8:2242. doi: 10.3389/fpls.2017.02242. 
            eCollection 2017.
  PUBMED    29449848
REFERENCE   2  (bases 1 to 5778)
  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 Plant
            Sci."; date: "2018-02-1"; volume: "8:2242. doi"; pages: " 
            10.3389/fpls.2017.02242. eCollection 2017"
COMMENT     Sequence Label: pUC18T-mini-Tn7T-Gm-Pc-DsRed-Express2
FEATURES             Location/Qualifiers
     source          1..5778
                     /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)."
     CDS             complement(2331..3008)
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MNSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(2393..2416)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2591..2610)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2808..2828
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     RBS             3016..3038
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     mobile_element  3295..3460
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3619..3727
                     /note="incP origin of transfer"
     rep_origin      complement(3959..4547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4039..4058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4718..5509,5510..5578))
                     /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     5341..5360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5579..5683)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5751..5769
                     /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 ctgttgctga ctcataccag gtagtccgag
     2041 gagctttatg cttgtaaacc gttttgtgaa aaaattttta aaataaaaaa ggggacctct
     2101 agggtcccca attaattagt aatataatct attaaaggtc attcaaaagg tcatccaccg
     2161 gatcaattcc cctgctcgcg caggctgggt gccaagctct cgggtaacat caaggcccga
     2221 tccttggagc ccttgccctc ccgcacgatg atcgtgccgt gatcgaaatc cagatccttg
     2281 acccgcagtt gcaaaccctc actgatccgc tagtccgagg cctcgagtta ctactggaac
     2341 aggtggtggc gggcctcggc gcgctcgtac tgctccacca cggtgtagtc ctcgttgtgg
     2401 gaggtgatgt ccagcttgga gtccacgtag tagtagccgg gcagcttcac gggcttcttg
     2461 gccatgtaga ttgacttgaa ctccaccagg tagtggccgc cgcccttcag cttcagcgcc
     2521 ttgtggatct cgcccttcag cacgccgtcg cgggggtaca ggcgctcggt ggagggctcc
     2581 cagcccagag tcttcttctg cattacgggg ccgtcggagg ggaagttcac gccgatgaac
     2641 ttcacgtggt agatgaaggt gccgtcctgc agggaggagt cctgggtcac ggtcaccacg
     2701 ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa ggacagcttc
     2761 ttgtagtcgg ggatgtcggc ggggtgcttc acgtacacct tggagccgta ctggaactgg
     2821 ggggacagga tgtcccaggc gaagggcagg gggccgccct tggtcacctg cagcttggcg
     2881 gtctgggtgc cctcgtaggg cttgccctcg ccctcgccct cgatctcgaa ctcgtggccg
     2941 ttcacggagc cctccatgtg caccttgaag cgcatgaagg gcttgatgac gttctcagtg
     3001 gaattcatat gtatatctcc ttcttaaagt taaacaaaat tatttcttga acgggttcgc
     3061 tacctgcatt acagcctaca gaccgagaag gcttatgtct actgggccaa ggcatttgtg
     3121 ttgtggacgg cccgcagcct tggtgggttt cgacatccgc gcgaaatggg gcaagctgaa
     3181 gtcgagggtt ttctgaccat gctcgccacc gagaagcaag tggcgccggc cacccaccgg
     3241 caggcgctca acgcgctgtt gttcttgtat cggctcgcga aggccttgca ggccaaccag
     3301 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
     3361 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
     3421 ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg
     3481 ttatgtttgg gcgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3541 tgggcgctct tccgcttcct cgctcactga cccgctgcgc tcggtcgttc ggctgcggcg
     3601 agcggtatca gagcttatcg gccagcctcg cagagcagga ttcccgttga gcaccgccag
     3661 gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat
     3721 cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa
     3781 tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa
     3841 gcagggttat gcagcggaaa gtatacctta aggaatcccc tgataacgca ggaaagaaca
     3901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcgagt cagaggtggc
     4021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4381 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4621 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4681 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4801 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4861 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4921 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5161 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5221 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5401 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5581 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5641 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5701 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5761 tcacgaggcc ctttcgtc
//
