LOCUS       sequence-209135-        5859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini-Tn7 vector for insertion of mTurquoise2 (codon optimized for P.
            fluorescens) at bacterial attTn7 site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5859)
  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 5859)
  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-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..5859
                     /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
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     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(2343..2360)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3097..3119
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     mobile_element  3376..3541
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3700..3808
                     /note="incP origin of transfer"
     rep_origin      complement(4040..4628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4120..4139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4799..5590,5591..5659))
                     /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     5422..5441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5660..5764)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5832..5850
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac 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 gcgtaagcgg 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 gttcctattc 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 caaacctggg
     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 gctagtccga
     2041 ggagctttat gcttgtaaac cgttttgtga aaaaattttt aaaataaaaa aggggacctc
     2101 tagggtcccc aattaattag taatataatc tattaaaggt cattcaaaag gtcatccacc
     2161 ggatcaattc ccctgctcgc gcaggctggg tgccaagctc tcgggtaaca tcaaggcccg
     2221 atccttggag cccttgccct cccgcacgat gatcgtgccg tgatcgaaat ccagatcctt
     2281 gacccgcagt tgcaaaccct cactgatccg ctagtccgag gcctcgagat gcttaaagct
     2341 tagtgatggt gatgatgatg tgcggccgct tacttgtaca gctcgtccat gcccagggtg
     2401 atgccggccg cggtcacgaa ctccagcaga accatgtggt cgcgtttttc gttaggatct
     2461 ttactgagtt tactttgggt ggagaggtaa tggttgtcag gcagcagaac ggggccgtcg
     2521 ccgatggggg tattttgttg ataatggtct gcgagttgaa caccgccgtc ttcgatatta
     2581 tggcggattt taaaattagc tttgatgccg tttttctgtt tgtcagcggt gatgtaaacg
     2641 ttgtcggaga aatagttgta ttcgagtttg tggcccagga tgttaccgtc ttccttaaaa
     2701 tcaatgccct tcagttcgat gcggttgacc agggtatcgc cttcgaattt cacttcggcc
     2761 cgagttttgt agttgccgtc gtctttgaag aaaatggtgc gttcttgaac atagccttct
     2821 ggcatggcgc ttttgaagaa atcgtgttgt ttcatatggt cggggtaacg ggcaaagcat
     2881 tggacgcccc agctcaaggt ggtcacgagg gtaggccaag gaacagggag tttgccggta
     2941 gtgcagataa atttcagggt cagtttgccg taagttgcat cgccctcgcc ttcgccgctc
     3001 acggaaaatt tatggccatt cacgtcgcca tcgagttcca cgaggattgg cacaacgccg
     3061 gtaaagagct cttcgccctt ggaattcata tgtatatctc cttcttaaag ttaaacaaaa
     3121 ttatttcttg aacgggttcg ctacctgcat tacagcctac agaccgagaa ggcttatgtc
     3181 tactgggcca aggcatttgt gttgtggacg gcccgcagcc ttggtgggtt tcgacatccg
     3241 cgcgaaatgg ggcaagctga agtcgagggt tttctgacca tgctcgccac cgagaagcaa
     3301 gtggcgccgg ccacccaccg gcaggcgctc aacgcgctgt tgttcttgta tcggctcgcg
     3361 aaggccttgc aggccaacca gataagtgaa atctagttcc aaactatttt gtcattttta
     3421 attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta
     3481 aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac
     3541 agcggggcat ttttcttcct gttatgtttg ggcgctgcat taatgaatcg gccaacgcgc
     3601 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg acccgctgcg
     3661 ctcggtcgtt cggctgcggc gagcggtatc agagcttatc ggccagcctc gcagagcagg
     3721 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
     3781 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
     3841 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
     3901 aatcgctata atgaccccga agcagggtta tgcagcggaa agtatacctt aaggaatccc
     3961 ctgataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4021 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4081 gacgctcgag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4141 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4201 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4261 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4321 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4381 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4441 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4501 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4561 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4621 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4681 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4741 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4801 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4861 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4921 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4981 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5041 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5101 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5161 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5221 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5281 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5341 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5401 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5461 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5521 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5581 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5641 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5701 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5761 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     5821 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
