LOCUS       sequence-408000-        5849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini-Tn7 vector for insertion of mNeonGreen (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 5849)
  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 5849)
  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-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            2052..2160
                     /note="incP origin of transfer"
     mobile_element  2319..2484
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     RBS             2741..2763
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3491..3508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3877..3924)
                     /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)."
     promoter        join(3965..3970,3971..3987,3988..3993)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4182..4715
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(4845..4892)
                     /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)."
     terminator      complement(4938..5024)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(5127..5221)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(5249..5265)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5250..5266)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5680..5699)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5799..5821
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac tcgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcagg ggattcctta aggtatactt
     1921 tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt
     1981 ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa
     2041 cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc
     2101 actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg
     2161 ataagctctg ataccgctcg ccgcagccga acgaccgagc gcagcgggtc agtgagcgag
     2221 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2281 tgcagcgccc aaacataaca ggaagaaaaa tgccccgctg tgggcggaca aaatagttgg
     2341 gaactgggag gggtggaaat ggagttttta aggattattt agggaagagt gacaaaatag
     2401 atgggaactg ggtgtagcgt cgtaagctaa tacgaaaatt aaaaatgaca aaatagtttg
     2461 gaactagatt tcacttatct ggttggcctg caaggccttc gcgagccgat acaagaacaa
     2521 cagcgcgttg agcgcctgcc ggtgggtggc cggcgccact tgcttctcgg tggcgagcat
     2581 ggtcagaaaa ccctcgactt cagcttgccc catttcgcgc ggatgtcgaa acccaccatg
     2641 gctgcgggcc gtccacaaca caaatgcctt ggcccagtag acataagcct tctcggtctg
     2701 taggctgtaa tgcaggtagc gaacccgttc tagaaataat tttgtttaac tttaagaagg
     2761 agatatacat atgaattcga aaggcgaaga agataacatg gccagtttac cggcaactca
     2821 tgaattacat atttttggaa gtattaacgg cgttgacttt gatatggttg gacagggcac
     2881 cggcaacccg aacgatggtt atgaagaatt aaacctgaaa tctacaaaag gcgacctgca
     2941 attctctccc tggattcttg ttccacacat tggttatggc ttccaccaat acttacccta
     3001 tcctgatggc atgtcccctt tccaagccgc gatggtggat ggctctggtt accaagtaca
     3061 ccgtaccatg caattcgaag atggagcatc actcacagta aactaccgct atacatatga
     3121 aggttcccac attaaaggcg aagcacaagt taaaggtact ggattccccg ctgatggacc
     3181 agttatgact aactcattga ccgcagcaga ctggtgtcgc tcaaaaaaaa cctatccgaa
     3241 cgataaaacg attattagca catttaaatg gagttatacg accggaaatg gcaaacgtta
     3301 ccgcagcaca gcacgcacaa cctatacatt tgcaaaacca atggccgcga attatttgaa
     3361 aaaccaaccg atgtatgtct ttcgtaaaac agaattaaaa cacagcaaaa cagaattaaa
     3421 ttttaaagaa tggcaaaaag catttaccga cgttatggga atggatgaat tatataaata
     3481 agcggccgca catcatcatc accatcacta agctttaagc atctcgaggc ctcggactag
     3541 cggatcagtg agggtttgca actgcgggtc aaggatctgg atttcgatca cggcacgatc
     3601 atcgtgcggg agggcaaggg ctccaaggat cgggccttga tgttacccga gagcttggca
     3661 cccagcctgc gcgagcaggg gaattgatcc ggtggatgac cttttgaatg acctttaata
     3721 gattatatta ctaattaatt ggggacccta gaggtcccct tttttatttt aaaaattttt
     3781 tcacaaaacg gtttacaagc ataaagctcc tcggactacc tggtatgagt cagcaacaga
     3841 gctcatgcat gatcgaatta gcttcaaaag cgctctgaag ttcctatact ttctagagaa
     3901 taggaacttc ggaataggaa cttcaagatc ccctgattcc ctttgtcaac agcaatggat
     3961 cgaattgaca taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg
     4021 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     4081 ggcttgttat gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     4141 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg
     4201 atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca
     4261 ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt
     4321 tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc
     4381 tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg
     4441 ttgttggcgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct
     4501 atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca
     4561 tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag
     4621 attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga
     4681 tgcactttga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct
     4741 tcccggccgc ggagttgttc ggtaaattgt cacaacgccg cggccaattc gatctagaat
     4801 tccattgagt aagtttttaa gcacatcagc ttcaaaagcg ctctgaagtt cctatacttt
     4861 ctagagaata ggaacttcgg aataggtact tcaagatccc caattcgatc gtccgggccg
     4921 caagctccta gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     4981 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct caagctagag
     5041 agtcattacc ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
     5101 ctgccactca tcgcagtcta gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     5161 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     5221 ccaagctcag ctaattaagc tagcttatcg ataccgtcga cctcgaaccc cacgcccctc
     5281 tttaatacga cgggcaattt gcacttcaga aaatgaagag tttgctttag ccataacaaa
     5341 agtccagtat gctttttcac agcataactg gactgatttc agtttacaac tattctgtct
     5401 agtttaagac tttattgtca tagtttagat ctattttgtt cagtttaaga ctttattgtc
     5461 cgcccacacc tgcttacgca gggcatccat ttattactca accgtaaccg attttgccag
     5521 gttacgcggc tggtcctcta gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5581 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     5641 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     5701 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5761 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5821 gttttcaccg tcgtcaccga aacgcgcga
//
