LOCUS       sequence-435259-        4911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini-Tn7 base vector with transcriptional terminator and oriT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4911)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer 
            RR, Schweizer HP
  TITLE     A Tn7-based broad-range bacterial cloning and expression system.
  JOURNAL   Nat Methods. 2005 Jun;2(6):443-8.
  PUBMED    15908923
REFERENCE   2  (bases 1 to 4911)
  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: "Nat 
            Methods."; date: "2005-06"; volume: "2(6)"; pages: "443-8"
COMMENT     Sequence Label: pUC18R6KT-mini-Tn7T-Km
FEATURES             Location/Qualifiers
     source          1..4911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  369..534
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     protein_bind    complement(649..696)
                     /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             1083..1877
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1137..1156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1747..1766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(1985..2032)
                     /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(2080..2166)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2269..2363)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2391..2407)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2392..2408)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2822..2841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2941..2963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3001..3019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3087..3191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3192..3260,3261..4052)
                     /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(3410..3429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4461..4817
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            join(4868..4911,1..65)
                     /note="incP origin of transfer"
ORIGIN
        1 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
       61 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
      121 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
      181 agggttatgc agcggaaagt ataccttaag gaatcccctg ataccgctcg ccgcagccga
      241 acgaccgagc gcagcgggtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
      301 cctctccccg cgcgttggcc gattcattaa tgcagcgccc aaacataaca ggaagaaaaa
      361 tgccccgctg tgggcggaca aaatagttgg gaactgggag gggtggaaat ggagttttta
      421 aggattattt agggaagagt gacaaaatag atgggaactg ggtgtagcgt cgtaagctaa
      481 tacgaaaatt aaaaatgaca aaatagtttg gaactagatt tcacttatct ggttggcctg
      541 caaggccttc gcgaggtacc gggcccaagc ttctcgagga attcctgcag cccgggggat
      601 ccactagtga gctcatgcat gatctcgaat tagcttcaaa agcgctctga agttcctata
      661 ctttctagag aataggaact tcggaatagg aacttcaaga tcccctgatt ccctttgtca
      721 acagcaatgg ataattcgat ttaacaaatg catggcgcaa gggctgctaa aggaagcgga
      781 acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
      841 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
      901 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
      961 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     1021 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc
     1081 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     1141 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     1201 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     1261 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     1321 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     1381 aggatctcct gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     1441 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     1501 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     1561 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     1621 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     1681 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     1741 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     1801 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     1861 ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagg atccaacatt tccaatcact
     1921 agtgaattat ctagaattat tccattgagt aagtttttaa gcacatcagc ttcaaaagcg
     1981 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggtact tcaagatccc
     2041 caattcgaga tcgtccgggc cgcaagctcc tagcggcgga tttgtcctac tcaggagagc
     2101 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
     2161 tatttgatgc ctcaagctag agagtcatta ccccaggcgt ttaagggcac caataactgc
     2221 cttaaaaaaa ttacgccccg ccctgccact catcgcagtc tagcttggat tctcaccaat
     2281 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     2341 tactggatct atcaacagga gtccaagctc agctaattaa gctagcttat cgataccgtc
     2401 gacctcgaac cccacgcccc tctttaatac gacgggcaat ttgcacttca gaaaatgaag
     2461 agtttgcttt agccataaca aaagtccagt atgctttttc acagcataac tggactgatt
     2521 tcagtttaca actattctgt ctagtttaag actttattgt catagtttag atctattttg
     2581 ttcagtttaa gactttattg tccgcctaca cccgcttacg cagggcatcc atttattact
     2641 caaccgtaac cgattttgcc aggttacgcg gctggtcctc tagctggcgt aatagcgaag
     2701 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     2761 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     2821 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     2881 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2941 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3001 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3061 caggtggcac ttttcgggga aatgtacgcg gaacccctat ttgtttattt ttctaaatac
     3121 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3181 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3241 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3301 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3361 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3421 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3481 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3541 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3601 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3661 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3721 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3781 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3841 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3901 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3961 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4021 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4081 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4441 taatcctgtt accagccggt tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt
     4501 tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta
     4561 aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt
     4621 agaggctatt taagttgctg atttatatta attttattgt tcaaacatga gagcttagta
     4681 cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt
     4741 ttagttcgtt aaacatgaga gcttagtacg ttaaacttga gagcttagta cgtgaaacat
     4801 gagagcttag tacgtactat caacaggttg aactgcccat gttctttcct gcgttatcag
     4861 agcttatcgg ccagcctcgc agagcaggat tcccgttgag caccgccagg t
//
