LOCUS       sequence-210896-        6694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniTn7 delivery plasmid with rhaSR-PrhaBAD inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6694)
  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 6694)
  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: pJM220 (pUC18T-miniTn7T-gm-rhaSR-PrhaBAD)
FEATURES             Location/Qualifiers
     source          1..6694
                     /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)"
     mobile_element  4211..4376
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            4535..4643
                     /note="incP origin of transfer"
     rep_origin      complement(4875..5463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4955..4974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5634..6425,6426..6494))
                     /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     6257..6276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6495..6599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6667..6685
                     /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 gaattcctcg agaagcttgg gcccggtacc tcgcgaaggc
     4201 cttgcaggcc aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt
     4261 cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa
     4321 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg
     4381 ggcatttttc ttcctgttat gtttgggcgc tgcattaatg aatcggccaa cgcgcgggga
     4441 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgacccg ctgcgctcgg
     4501 tcgttcggct gcggcgagcg gtatcagagc ttatcggcca gcctcgcaga gcaggattcc
     4561 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     4621 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     4681 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     4741 ctataatgac cccgaagcag ggttatgcag cggaaagtat accttaagga atcccctgat
     4801 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4861 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4921 tcgagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4981 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5041 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5101 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5161 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5221 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5281 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5341 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5401 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5461 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5521 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5581 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5641 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5701 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5761 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5821 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5881 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5941 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6001 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6061 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6121 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6181 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6241 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6301 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6361 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6421 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6481 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6541 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6601 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6661 tataaaaata ggcgtatcac gaggcccttt cgtc
//
