LOCUS       sequence-417531-        7535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pQF derivative with promoter PQ5 exchanged for promoter PQJ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7535)
  AUTHORS   Klotz A, Kaczmarczyk A, Jenal U
  TITLE     A Synthetic Cumate-Inducible Promoter for Graded and Homogenous Gene
            Expression in Pseudomonas aeruginosa.
  JOURNAL   Appl Environ Microbiol. 2023 May 18:e0021123. doi: 
            10.1128/aem.00211-23.
  PUBMED    37199671
REFERENCE   2  (bases 1 to 7535)
  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: "Appl 
            Environ Microbiol. 2023 May 18:e0021123. doi: 10.1128/aem.00211-23."
COMMENT     Sequence Label: pQFT
FEATURES             Location/Qualifiers
     source          1..7535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      24..655
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1118..1706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1607..1626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1768..2379)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    2456..2483
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    2526..2553
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     CDS             2578..2643
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2698..2763
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2835..2852)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2835..2851)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2844..2866)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3060..3079)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3179..3201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3239..3257)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             4043..5242
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(5473..6621)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(6893..7264)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7297..7406
                     /note="incP origin of transfer"
ORIGIN
        1 gaaaagatcc gtcgaccctt tccgacgctc accgggctgg ttgccctcgc cgctgggctg
       61 gcggccgtct atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac
      121 cgcggccgcc ggcgttgtgg atacctcgcg gaaaacttgg ccctcactga cagatgaggg
      181 gcggacgttg acacttgagg ggccgactca cccggcgcgg cgttgacaga tgaggggcag
      241 gctcgatttc ggccggcgac gtggagctgg ccagcctcgc aaatcggcga aaacgcctga
      301 ttttacgcga gtttcccaca gatgatgtgg acaagcctgg ggataagtgc cctgcggtat
      361 tgacacttga ggggcgcgac tactgacaga tgaggggcgc gatccttgac acttgagggg
      421 cagagtgctg acagatgagg ggcgcaccta ttgacatttg aggggctgtc cacaggcaga
      481 aaatccagca tttgcaaggg tttccgcccg tttttcggcc accgctaacc tgtcttttaa
      541 cctgctttta aaccaatatt tataaacctt gtttttaacc agggctgcgc cctgtgcgcg
      601 tgaccgcgca cgccgaaggg gggtgccccc ccttctcgaa ccctcccggc ccgctaacgc
      661 gggcctccca tccccccagg ggctgcgccc ctcggccgcg aacggcctca ccccaaaaat
      721 ggcagccaag ctgaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
      781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
      841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
      901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
      961 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1081 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1201 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1261 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1321 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1381 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1441 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1501 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1561 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1621 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1681 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1741 ccttttgctg gccttttgct cacatgttca gcgcttgaac ttggcgtagc gctcgcgggc
     1801 gatttccagg gtcgagttgc ggacgcgctc gaagcgctcc ttgtccttct gccacagcga
     1861 gcggaccgcc aggccgcgga ccgagttgaa gatcagccac aggatgtcct ccgcgtcgtc
     1921 gcggctcagg ccgcgcgaca ccagcacgcc cagccacata tcctcgacga cgaagcggtt
     1981 gcgctcgacg gtgcgctgga tgccctcgcg cagcgccggg tcgcggtcgg cggccacgat
     2041 caggtccagc gagatcgaga agtcgtcgtc caggaagaac tcggcggcgt cgtccagcat
     2101 ctgctggatg acgtcgtcct cgggcttcag cttggccagg cgggcgcggc tgcgctcggt
     2161 gatctgctcg tacagccact cgaaggtggc cagcagcagt tccagcttgg tcgggaagtg
     2221 gtgcgactgg gcgccgcggc tgacgccggc ggcgcccggg acgtcggcga tgcggaagcc
     2281 ggcatagccc ttctcgcgca ggacgcccag ggcggcggcg atcagcttgc cctgggtttc
     2341 catcgcgcgc tcggcctggg tgcggcgctt cggcgacatg atgaccattc ggggtctctc
     2401 cctgctagca tcagggttat tgtatcatga gcggatacat atgtcaatgt accggaacaa
     2461 acagacaatc tggtctgttt gtacaatatt gacgccctgt gggggcatcc gtatgattgc
     2521 gcctcaacaa acagacaatc tggtctgttt gtaactagta gaggaagctt ccgcatggac
     2581 tacaaggacc acgacggcga ctataaggac cacgacatcg actacaagga tgacgacgac
     2641 aagcttatta atatgactct agaagatctc gaggtaccga gctccaattg ggatccggac
     2701 tacaaggacc acgacggcga ctataaggac cacgacatcg actacaagga tgacgacgac
     2761 aagtgaggat ccggtgaagt gacccgcaaa aaaaaaggcc aggggatcgc tcccccggcc
     2821 ttttttttga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2881 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2941 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     3001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     3061 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3121 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3181 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3241 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagcaccct
     3301 ttctcggtcc ttcaacgttc ctgacaacga gcctcctttt cgccaatcca tcgacaatca
     3361 ccgcgagtcc ctgctcgaac gctgcgtccg gaccggcttc gtcgaaggcg tctatcgcgg
     3421 cccgcaacag cggcgagagc ggagcctgtt caacggtgcc gccgcgctcg ccggcatcgc
     3481 tgtcgccggc ctgctcctca agcacggccc caacagtgaa gtagctgatt gtcatcagcg
     3541 cattgacggc gtccccggcc gaaaaacccg cctcgcagag gaagcgaagc tgcgcgtcgg
     3601 ccgtttccat ctgcggtgcg cccggtcgcg tgccggcatg gatgcgcgcg ccatcgcggt
     3661 aggcgagcag cgcctgcctg aagctgcggg cattcccgat cagaaatgag cgccagtcgt
     3721 cgtcggctct cggcaccgaa tgcgtatgat tctccgccag catggcttcg gccagtgcgt
     3781 cgagcagcgc ccgcttgttc ctgaagtgcc agtaaagcgc cggctgctga acccccaacc
     3841 gttccgccag tttgcgtgtc gtcagaccgt ctacgccgac ctcgttcaac aggtccaggg
     3901 cggcacggat cactgtattc ggctgcaact ttgtcatgct tgacacttta tcactgataa
     3961 acataatatg tccaccaact tatcagtgat aaagaatccg cgcgttcaat cggaccagcg
     4021 gaggctggtc cggaggccag acgtgaaacc caacataccc ctgatcgtaa ttctgagcac
     4081 tgtcgcgctc gacgctgtcg gcatcggcct gattatgccg gtgctgccgg gcctcctgcg
     4141 cgatctggtt cactcgaacg acgtcaccgc ccactatggc attctgctgg cgctgtatgc
     4201 gttggtgcaa tttgcctgcg cacctgtgct gggcgcgctg tcggatcgtt tcgggcggcg
     4261 gccaatcttg ctcgtctcgc tggccggcgc cactgtcgac tacgccatca tggcgacagc
     4321 gcctttcctt tgggttctct atatcgggcg gatcgtggcc ggcatcaccg gggcgactgg
     4381 ggcggtagcc ggcgcttata ttgccgatat cactgatggc gatgagcgcg cgcggcactt
     4441 cggcttcatg agcgcctgtt tcgggttcgg gatggtcgcg ggacctgtgc tcggtgggct
     4501 gatgggcggt ttctcccccc acgctccgtt cttcgccgcg gcagccttga acggcctcaa
     4561 tttcctgacg ggctgtttcc ttttgccgga gtcgcacaaa ggcgaacgcc ggccgttacg
     4621 ccgggaggct ctcaacccgc tcgcttcgtt ccggtgggcc cggggcatga ccgtcgtcgc
     4681 cgccctgatg gcggtcttct tcatcatgca acttgtcgga caggtgccgg ccgcgctttg
     4741 ggtcattttc ggcgaggatc gctttcactg ggacgcgacc acgatcggca tttcgcttgc
     4801 cgcatttggc attctgcatt cactcgccca ggcaatgatc accggccctg tagccgcccg
     4861 gctcggcgaa aggcgggcac tcatgctcgg aatgattgcc gacggcacag gctacatcct
     4921 gcttgccttc gcgacacggg gatggatggc gttcccgatc atggtcctgc ttgcttcggg
     4981 tggcatcgga atgccggcgc tgcaagcaat gttgtccagg caggtggatg aggaacgtca
     5041 ggggcagctg caaggctcac tggcggcgct caccagcctg acctcgatcg tcggacccct
     5101 cctcttcacg gcgatctatg cggcttctat aacaacgtgg aacgggtggg catggattgc
     5161 aggcgctgcc ctctacttgc tctgcctgcc ggcgctgcgt cgcgggcttt ggagcggcgc
     5221 agggcaacga gccgatcgct gatcgtggaa acgataggcc tatgccatgc gggtcaaggc
     5281 gacttccggc aagctatacg cgccctagaa ttgtcaattt taatcctctg tttatcggca
     5341 gttcgtagag cgcgccgtgc gtcccgagcg atactgagcg aagcaagtgc gtcgagcagt
     5401 gcccgcttgt tcctgaaatg ccagtaaagc gctggctgct gaacccccag ccggaactga
     5461 ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac
     5521 caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac
     5581 gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc
     5641 ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt
     5701 ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat
     5761 caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag
     5821 cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag
     5881 gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc
     5941 ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta
     6001 ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta
     6061 ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg
     6121 gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg
     6181 catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg
     6241 cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg ccaggtcctc
     6301 gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt
     6361 cgccaaacct gccgcctcct gttcgagacg acgcgaacgc tccacggcgg ccgatggcgc
     6421 gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg
     6481 gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc
     6541 gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt
     6601 ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc acgccggggc
     6661 aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat aatccacctt
     6721 atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact tggtattccg
     6781 aatcttgccc tgcacgaata ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg
     6841 gacgtgcttg gcaatcacgc gcaccccccg gccgttttag cggctaaaaa agtcatggct
     6901 ctgccctcgg gcggaccacg cccatcatga ccttgccaag ctcgtcctgc ttctcttcga
     6961 tcttcgccag cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg
     7021 tgagccagag tttcagcagg ccgcccaggc ggcccaggtc gccattgatg cgggccagct
     7081 cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca
     7141 ggtaggccta caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt
     7201 ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag
     7261 ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat
     7321 tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg
     7381 gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta
     7441 cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa
     7501 tcgctataat gaccccgaag cagggttatg cagcg
//
