LOCUS       sequence-418766-        5723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GmR mini-Tn7 vector, mobilizable, for RFP tagging bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5723)
  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 5723)
  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-Gm-dsRedExpress
FEATURES             Location/Qualifiers
     source          1..5723
                     /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
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     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)."
     terminator      complement(2099..2185)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2288..2382)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2437..3114)
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2499..2522)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     2914..2934
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     RBS             3121..3132
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3151..3167
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    3184..3200
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     mobile_element  3240..3405
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3564..3672
                     /note="incP origin of transfer"
     rep_origin      complement(3904..4492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3984..4003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4663..5454,5455..5523))
                     /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     5286..5305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5524..5628)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5696..5714
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgacgac 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 caaacttggg
     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 tcccccgggc tgcaggaatt
     2041 cctcgagaag cttgggcccg gtacctcgcg aaggcgggcc gcaagctcct agcggcggat
     2101 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2161 tcgactgagc ctttcgtttt atttgatgcc tcaagctaga gagtcattac cccaggcgtt
     2221 taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtct
     2281 agcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     2341 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagctca gctaattaag
     2401 cttggtaccg gccgccagtg tgctggaatt cggcttctac aggaacaggt ggtggcggcc
     2461 ctcggtgcgc tcgtactgct ccacgatggt gtagtcctcg ttgtgggagg tgatgtccag
     2521 cttggcgtcc acgtagtagt agccgggcag ctgcacgggc ttcttggcca tgtagatgga
     2581 cttgaactcc accaggtagt ggccgccgtc cttcagcttc agggccttgt gggtctcgcc
     2641 cttcagcacg ccgtcgcggg ggtacaggcg ctcggtggag gcctcccagc ccatggtctt
     2701 cttctgcatc acggggccgt cggaggggaa gttcacgccg atgaacttca ccttgtagat
     2761 gaagcagccg tcctgcaggg aggagtcctg ggtcacggtc gccacgccgc cgtcctcgaa
     2821 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcttgt agtcggggat
     2881 gtcggcgggg tgcttcacgt acaccttgga gccgtactgg aactgggggg acaggatgtc
     2941 ccaggcgaag ggcagggggc cgcccttggt caccttcagc ttcacggtgt tgtggccctc
     3001 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggtgccctc
     3061 catgcgcacc ttgaagcgca tgaactcggt gatgacgttc tcggaggagg ccatgcttaa
     3121 tttctcctct ttaattctag atgtgtgaaa ttgttatccg ctcacaattg aatctaagta
     3181 tcattgttat ccgctcacaa gtcaacactc tttttgataa attttctccc ttgcaggcca
     3241 accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt
     3301 acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac
     3361 tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct
     3421 tcctgttatg tttgggcgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3481 cgtattgggc gctcttccgc ttcctcgctc actgacccgc tgcgctcggt cgttcggctg
     3541 cggcgagcgg tatcagagct tatcggccag cctcgcagag caggattccc gttgagcacc
     3601 gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca
     3661 cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg
     3721 caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc
     3781 ccgaagcagg gttatgcagc ggaaagtata ccttaaggaa tcccctgata acgcaggaaa
     3841 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3901 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct cgagtcagag
     3961 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4021 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4081 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4141 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4201 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4261 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4321 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4381 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4441 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4501 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4561 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4621 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4681 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4741 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4801 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4861 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4921 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4981 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5041 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5101 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5161 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5221 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5281 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5341 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5401 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5461 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5581 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5641 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5701 gcgtatcacg aggccctttc gtc
//
