LOCUS       sequence-210981-        7231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniTn7 delivery plasmid with rhaSR-PrhaBAD inducible promoter and 
            StRBS-aacC1 insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7231)
  AUTHORS   Meisner J, Goldberg JB
  TITLE     The Escherichia coli rhaSR-PrhaBAD Inducible Promoter System Allows 
            Tightly Controlled Gene Expression over a Wide Range in Pseudomonas 
            aeruginosa.
  JOURNAL   Appl Environ Microbiol. 2016 Oct 27;82(22):6715-6727. doi: 
            10.1128/AEM.02041-16. Print 2016 Nov 15.
  PUBMED    27613678
REFERENCE   2  (bases 1 to 7231)
  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"; doi: 
            "10.1128/AEM.02041-16"; journalName: "Appl Environ Microbiol"; date:
            "2016-10-27- 27"; volume: "82"; issue: "22"; pages: "6715-6727"
COMMENT     Sequence Label: pJM240
            (pUC18T-miniTn7T-gm-rhaSR-PrhaBAD-stRBS...
FEATURES             Location/Qualifiers
     source          1..7231
                     /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)."
     CDS             complement(2113..2961)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(3035..3871)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        4017..4135
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             4176..4709
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4748..4913
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            5072..5180
                     /note="incP origin of transfer"
     rep_origin      complement(5412..6000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5492..5511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6171..6962,6963..7031))
                     /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     6794..6813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7032..7136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7204..7222
                     /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 caaagagtgg aacaatgcag gacgccgtcc
     2041 ggtaacggtg ttgagcctcg gttgtgtggt gattattgtc gccgctaaca tcgtcggcat
     2101 cggcatggcg aattaatctt tctgcgaatt gagatgacgc cactggctgg gcgtcatccc
     2161 ggtttcccgg gtaaacacca ccgaaaaata gttactatct tcaaagccac attcggtcga
     2221 aatatcactg attaacaggc ggctatgctg gagaagatat tgcgcatgac acactctgac
     2281 ctgtcgcaga tattgattga tggtcattcc agtctgctgg cgaaattgct gacgcaaaac
     2341 gcgctcactg cacgatgcct catcacaaaa tttatccagc gcaaagggac ttttcaggct
     2401 agccgccagc cgggtaatca gcttatccag caacgtttcg ctggatgttg gcggcaacga
     2461 atcactggtg taacgatggc gattcagcaa catcaccaac tgcccgaaca gcaactcagc
     2521 catttcgtta gcaaacggca catgctgact actttcatgc tcaagctgac cgataacctg
     2581 ccgcgcctgc gccatcccca tgctacctaa gcgccagtgt ggttgccctg cgctggcgtt
     2641 aaatcccgga atcgccccct gccagtcaag attcagcttc agacgctccg ggcaataaat
     2701 aatattctgc aaaaccagat cgttaacgga agcgtaggag tgtttatcgt cagcatgaat
     2761 gtaaaagaga tcgccacggg taatgcgata agggcgatcg ttgagtacat gcaggccatt
     2821 accgcgccag acaatcacca gctcacaaaa atcatgtgta tgttcagcaa agacatcttg
     2881 cggataacgg tcagccacag cgactgcctg ctggtcgctg gcaaaaaaat catctttgag
     2941 aagttttaac tgatgcgcca ccgtggctac ctcggccaga gaacgaagtt gattattcgc
     3001 aatatggcgt acaaatacgt tgagaagatt cgcgttattg cagaaagcca tcccgtccct
     3061 ggcgaatatc acgcggtgac cagttaaact ctcggcgaaa aagcgtcgaa aagtggttac
     3121 tgtcgctgaa tccacagcga taggcgatgt cagtaacgct ggcctcgctg tggcgtagca
     3181 gatgtcgggc tttcatcagt cgcaggcggt tcaggtatcg ctgaggcgtc agtcccgttt
     3241 gctgcttaag ctgccgatgt agcgtacgca gtgaaagaga aaattgatcc gccacggcat
     3301 cccaattcac ctcatcggca aaatggtcct ccagccaggc cagaagcaag ttgagacgtg
     3361 atgcgctgtt ttccaggttc tcctgcaaac tgcttttacg cagcaagagc agtaattgca
     3421 taaacaagat ctcgcgactg gcggtcgagg gtaaatcatt ttccccttcc tgctgttcca
     3481 tctgtgcaac cagctgtcgc acctgctgca atacgctgtg gttaacgcgc cagtgagacg
     3541 gatactgccc atccagctct tgtggcagca actgattcag cccggcgaga aactgaaatc
     3601 gatccggcga gcgatacagc acattggtca gacacagatt atcggtatgt tcatacagat
     3661 gccgatcatg atcgcgtacg aaacagaccg tgccaccggt gatggtatag ggctgcccat
     3721 taaacacatg aatacccgtg ccatgttcga caatcacaat ttcatgaaaa tcatgatgat
     3781 gttcaggaaa atccgcctgc gggagccggg gttctatcgc cacggacgcg ttaccagacg
     3841 gaaaaaaatc cacactatgt aatacggtca tactggcctc ctgatgtcgt caacacggcg
     3901 aaatagtaat cacgaggtca ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa
     3961 acgtcgattt ttcaagatac agcgtgaatt ttcaggaaat gcggtgagca tcacatcacc
     4021 acaattcagc aaattgtgaa catcatcacg ttcatctttc cctggttgcc aatggcccat
     4081 tttcctgtca gtaacgagaa ggtcgcgaat tcaggcgctt tttagactgg tcgtaatgaa
     4141 attcaactag tgctctgcag taaggaggag caacgatgtt acgcagcagc aacgatgtta
     4201 cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca
     4261 catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc
     4321 gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga
     4381 acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg
     4441 gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct
     4501 atgatctcgc agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc
     4561 tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg
     4621 gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact
     4681 ttgatatcga cccaagtacc gccacctaaa agcttgggcc cggtacctcg cgaaggcctt
     4741 gcaggccaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     4801 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     4861 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc
     4921 atttttcttc ctgttatgtt tgggcgctgc attaatgaat cggccaacgc gcggggagag
     4981 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgacccgctg cgctcggtcg
     5041 ttcggctgcg gcgagcggta tcagagctta tcggccagcc tcgcagagca ggattcccgt
     5101 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc
     5161 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa
     5221 ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta
     5281 taatgacccc gaagcagggt tatgcagcgg aaagtatacc ttaaggaatc ccctgataac
     5341 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctcg
     5461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7201 aaaaataggc gtatcacgag gccctttcgt c
//
