LOCUS       sequence-109992-        4362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TpR mini-Tn7 vector, mobilizable.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4362)
  AUTHORS   Choi KH, Schweizer HP
  TITLE     mini-Tn7 insertion in bacteria with single attTn7 sites: example 
            Pseudomonas aeruginosa.
  JOURNAL   Nat Protoc. 2006;1(1):153-61.
  PUBMED    17406227
REFERENCE   2  (bases 1 to 4362)
  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 
            Protoc."; date: "2006"; volume: "1(1)"; pages: "153-61"
COMMENT     Sequence Label: pUC18T-mini-Tn7T-Tp
FEATURES             Location/Qualifiers
     source          1..4362
                     /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    963..1010
                     /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(1072..1308)
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     promoter        complement(join(1633..1638,1639..1655,1656..1661))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1713..1760
                     /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)."
     mobile_element  1879..2044
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            2203..2311
                     /note="incP origin of transfer"
     rep_origin      complement(2543..3131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2623..2642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3302..4093,4094..4162))
                     /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     3925..3944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4163..4267)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4335..4353
                     /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 atgagctcga attggggatc
      961 ttgaagtacc tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt
     1021 tgaagctgat gtgcttaaaa acttactcaa tggaataatt ctagataatt cttaggccac
     1081 acgttcaagt gcagccacag gataaatttg cactgagcct gggtgggatt cggactcgac
     1141 cgcatagcct tcaggagtga gttttgtgca ataccaaccg acgacttgac cctgccaagc
     1201 ggcaccagat ttcttgcgta cgcgatcccc taagccaaag gtggcactca ggggaagcgc
     1261 aaactgccct gcaacgggag cgttggcttc atcgctactt tgacccatgt cgaatccttc
     1321 ttgtgaatct attatggcga caagcaaatc gagctctgac tgcctacccc acaacaacta
     1381 tcagaaagca ccagcacaac ggctgcctaa ctttgtttta gggcgactgc cctgctgcgt
     1441 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
     1501 ggatgcccga ggctagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact
     1561 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
     1621 tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggttcg tgaattatcc
     1681 attgctgttg acaaagggaa tcaggggatc ttgaagttcc tattccgaag ttcctattct
     1741 ctagaaagta taggaacttc agagcgcttt tgaagctaat tcgagctcga tcatgcatga
     1801 gctcactagt ggatcccccg ggctgcagga attcctcgag aagcttgggc ccggtacctc
     1861 gcgaaggcct tgcaggccaa ccagataagt gaaatctagt tccaaactat tttgtcattt
     1921 ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc
     1981 ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc
     2041 cacagcgggg catttttctt cctgttatgt ttgggcgctg cattaatgaa tcggccaacg
     2101 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgacccgct
     2161 gcgctcggtc gttcggctgc ggcgagcggt atcagagctt atcggccagc ctcgcagagc
     2221 aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct
     2281 cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa
     2341 gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga
     2401 aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaagtatac cttaaggaat
     2461 cccctgataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2581 atcgacgctc gagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2701 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2941 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3061 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3121 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3181 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3241 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3301 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3361 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3421 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3481 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3541 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3601 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3661 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3721 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3781 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3841 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3901 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3961 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4021 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4081 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4141 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4201 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4261 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4321 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
