LOCUS       sequence-121755-        4967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini-Tn7 vector with oriR6K, for attTn7 site determination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4967)
  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 4967)
  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: pUC18T-mini-Tn7T-Gm-REP
FEATURES             Location/Qualifiers
     source          1..4967
                     /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)."
     rep_origin      2070..2426
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     mobile_element  2484..2649
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            2808..2916
                     /note="incP origin of transfer"
     rep_origin      complement(3148..3736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3228..3247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3907..4698,4699..4767))
                     /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     4530..4549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4768..4872)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4940..4958
                     /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 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 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 cactagtgga tccactagta acggccgcca
     2041 gtgtgctgga attcggctta ttaaccggtt gtcagccgtt aagtgttcct gtgtcactca
     2101 aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc ttgttgtcca
     2161 caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc ttataaaact
     2221 taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt caaacatgag
     2281 agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta gtacgttagc
     2341 catgagggtt tagttcgtta aacatgagag cttagtacgt taaacttgag agcttagtac
     2401 gtgaaacatg agagcttagt acgtactatc aacaggttga actgccgcgg taataagccg
     2461 aattctgcag atccttgcag gccaaccaga taagtgaaat ctagttccaa actattttgt
     2521 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     2581 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     2641 ccgcccacag cggggcattt ttcttcctgt tatgtttggg cgctgcatta atgaatcggc
     2701 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2761 ccgctgcgct cggtcgttcg gctgcggcga gcggtatcag agcttatcgg ccagcctcgc
     2821 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
     2881 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
     2941 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
     3001 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaag tataccttaa
     3061 ggaatcccct gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3721 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3781 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3841 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3901 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3961 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4021 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4081 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4141 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4201 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4261 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4321 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4381 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4441 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4501 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4561 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4621 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4681 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4741 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4801 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4861 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4921 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
