LOCUS       sequence-121765-        5519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TpR mini-Tn7 vector, mobilizable, for RFP tagging bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5519)
  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 5519)
  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-dsRedExpress
FEATURES             Location/Qualifiers
     source          1..5519
                     /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(1634..1639,1640..1656,1657..1662))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1714..1761
                     /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(1893..1979)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2082..2176)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2231..2908)
                     /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(2293..2316)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     2708..2728
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     RBS             2915..2926
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2945..2961
                     /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    2978..2994
                     /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  3036..3201
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            3360..3468
                     /note="incP origin of transfer"
     rep_origin      complement(3700..4288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3780..3799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4459..5250,5251..5319))
                     /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     5082..5101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5320..5424)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5492..5510
                     /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 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 ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac
     1561 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     1621 ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgaattatc
     1681 cattgctgtt gacaaaggga atcaggggat cttgaagttc ctattccgaa gttcctattc
     1741 tctagaaagt ataggaactt cagagcgctt ttgaagctaa ttcgagctcg atcatgcatg
     1801 agctcactag tggatccccc gggctgcagg aattcctcga gaagcttggg cccggtacct
     1861 cgcgaaggcg ggccgcaagc tcctagcggc ggatttgtcc tactcaggag agcgttcacc
     1921 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
     1981 tgcctcaagc tagagagtca ttaccccagg cgtttaaggg caccaataac tgccttaaaa
     2041 aaattacgcc ccgccctgcc actcatcgca gtctagcttg gattctcacc aataaaaaac
     2101 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
     2161 tctatcaaca ggagtccaag ctcagctaat taagcttggt accggccgcc agtgtgctgg
     2221 aattcggctt ctacaggaac aggtggtggc ggccctcggt gcgctcgtac tgctccacga
     2281 tggtgtagtc ctcgttgtgg gaggtgatgt ccagcttggc gtccacgtag tagtagccgg
     2341 gcagctgcac gggcttcttg gccatgtaga tggacttgaa ctccaccagg tagtggccgc
     2401 cgtccttcag cttcagggcc ttgtgggtct cgcccttcag cacgccgtcg cgggggtaca
     2461 ggcgctcggt ggaggcctcc cagcccatgg tcttcttctg catcacgggg ccgtcggagg
     2521 ggaagttcac gccgatgaac ttcaccttgt agatgaagca gccgtcctgc agggaggagt
     2581 cctgggtcac ggtcgccacg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc
     2641 cctcggggaa ggacagcttc ttgtagtcgg ggatgtcggc ggggtgcttc acgtacacct
     2701 tggagccgta ctggaactgg ggggacagga tgtcccaggc gaagggcagg gggccgccct
     2761 tggtcacctt cagcttcacg gtgttgtggc cctcgtaggg gcggccctcg ccctcgccct
     2821 cgatctcgaa ctcgtggccg ttcacggtgc cctccatgcg caccttgaag cgcatgaact
     2881 cggtgatgac gttctcggag gaggccatgc ttaatttctc ctctttaatt ctagatgtgt
     2941 gaaattgtta tccgctcaca attgaatcta agtatcattg ttatccgctc acaagtcaac
     3001 actctttttg ataaattttc tcgaccttgc aggccaacca gataagtgaa atctagttcc
     3061 aaactatttt gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc
     3121 tattttgtca ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc
     3181 caactatttt gtccgcccac agcggggcat ttttcttcct gttatgtttg ggcgctgcat
     3241 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3301 tcgctcactg acccgctgcg ctcggtcgtt cggctgcggc gagcggtatc agagcttatc
     3361 ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg
     3421 aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt
     3481 tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa
     3541 aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa
     3601 agtatacctt aaggaatccc ctgataacgc aggaaagaac atgtgagcaa aaggccagca
     3661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3721 tgacgagcat cacaaaaatc gacgctcgag tcagaggtgg cgaaacccga caggactata
     3781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4141 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
