LOCUS       sequence-109983-        5506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TpR mini-Tn7 vector, mobilizable, for GFP tagging bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5506)
  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 5506)
  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-gfpmut3
FEATURES             Location/Qualifiers
     source          1..5506
                     /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)."
     terminator      complement(1892..1978)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2081..2175)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2198..2911)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2238..2258)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2853..2881
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             2918..2929
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     mobile_element  3023..3188
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3347..3455
                     /note="incP origin of transfer"
     rep_origin      complement(3687..4275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3767..3786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4446..5237,5238..5306))
                     /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     5069..5088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5307..5411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5479..5497
                     /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 gcgaaggcgg gccgcaagct cctagcggcg gatttgtcct actcaggaga gcgttcaccg
     1921 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     1981 gcctcaagct agagagtcat taccccaggc gtttaagggc accaataact gccttaaaaa
     2041 aattacgccc cgccctgcca ctcatcgcag tctagcttgg attctcacca ataaaaaacg
     2101 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     2161 ctatcaacag gagtccaagc tcagctaatt aagcttattt gtatagttca tccatgccat
     2221 gtgtaatccc agcagctgtt acaaactcaa gaaggaccat gtggtctctc ttttcgttgg
     2281 gatctttcga aagggcagat tgtgtggaca ggtaatggtt gtctggtaaa aggacagggc
     2341 catcgccaat tggagtattt tgttgataat ggtctgctag ttgaacgctt ccatcttcaa
     2401 tgttgtgtct aattttgaag ttaactttga ttccattctt ttgtttgtct gccatgatgt
     2461 atacattgtg tgagttatag ttgtattcca atttgtgtcc aagaatgttt ccatcttctt
     2521 taaaatcaat accttttaac tcgattctat taacaagggt atcaccttca aacttgactt
     2581 cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat agttctttcc tgtacataac
     2641 cttcgggcat ggcactcttg aaaaagtcat gccgtttcat atgatctggg tatctcgcaa
     2701 agcattgaac accataaccg aaagtagtga caagtgttgg ccatggaaca ggtagttttc
     2761 cagtagtgca aataaattta agggtaagtt ttccgtatgt tgcatcacct tcaccctctc
     2821 cactgacaga aaatttgtgc ccattaacat caccatctaa ttcaacaaga attgggacaa
     2881 ctccagtgaa aagttcttct cctttacgca tgcttaattt ctcctcttta attctagagt
     2941 ggtgtcgcat tatagggagt tattccggcc tgactagagg aaatttaaaa taattttctg
     3001 accgcgcaac gccttgcagg ccaaccagat aagtgaaatc tagttccaaa ctattttgtc
     3061 atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc
     3121 ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc
     3181 cgcccacagc ggggcatttt tcttcctgtt atgtttgggc gctgcattaa tgaatcggcc
     3241 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgacc
     3301 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcaga gcttatcggc cagcctcgca
     3361 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
     3421 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
     3481 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
     3541 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaagt ataccttaag
     3601 gaatcccctg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3721 aaaaatcgac gctcgagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4381 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4441 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4501 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4561 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4621 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4681 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4741 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4801 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4861 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4921 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4981 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5041 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5101 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5161 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5221 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5281 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5401 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5461 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
