LOCUS       sequence-417525-        8938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pQFT-mNG with constitutively expressed mScarlet-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8938)
  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 8938)
  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-mNG-Pcon-Scar
FEATURES             Location/Qualifiers
     source          1..8938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1083..1671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1572..1591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1733..2344)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    2421..2448
                     /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    2491..2518
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     CDS             4066..4131
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4203..4220)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4203..4219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4212..4234)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4428..4447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4547..4569
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4607..4625)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             5411..6610
                     /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(6841..7989)
                     /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(8261..8632)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            8665..8774
                     /note="incP origin of transfer"
     rep_origin      join(8927..8938,1..620)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gctggttgcc ctcgccgctg ggctggcggc cgtctatggc cctgcaaacg cgccagaaac
       61 gccgtcgaag ccgtgtgcga gacaccgcgg ccgccggcgt tgtggatacc tcgcggaaaa
      121 cttggccctc actgacagat gaggggcgga cgttgacact tgaggggccg actcacccgg
      181 cgcggcgttg acagatgagg ggcaggctcg atttcggccg gcgacgtgga gctggccagc
      241 ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc ccacagatga tgtggacaag
      301 cctggggata agtgccctgc ggtattgaca cttgaggggc gcgactactg acagatgagg
      361 ggcgcgatcc ttgacacttg aggggcagag tgctgacaga tgaggggcgc acctattgac
      421 atttgagggg ctgtccacag gcagaaaatc cagcatttgc aagggtttcc gcccgttttt
      481 cggccaccgc taacctgtct tttaacctgc ttttaaacca atatttataa accttgtttt
      541 taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg cccccccttc
      601 tcgaaccctc ccggcccgct aacgcgggcc tcccatcccc ccaggggctg cgcccctcgg
      661 ccgcgaacgg cctcacccca aaaatggcag ccaagctgac cacttctgcg ctcggccctt
      721 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      781 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      841 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      901 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      961 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1021 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1081 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1141 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1201 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1261 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1321 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1381 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1441 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1501 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1561 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1621 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1681 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttcagcgct
     1741 tgaacttggc gtagcgctcg cgggcgattt ccagggtcga gttgcggacg cgctcgaagc
     1801 gctccttgtc cttctgccac agcgagcgga ccgccaggcc gcggaccgag ttgaagatca
     1861 gccacaggat gtcctccgcg tcgtcgcggc tcaggccgcg cgacaccagc acgcccagcc
     1921 acatatcctc gacgacgaag cggttgcgct cgacggtgcg ctggatgccc tcgcgcagcg
     1981 ccgggtcgcg gtcggcggcc acgatcaggt ccagcgagat cgagaagtcg tcgtccagga
     2041 agaactcggc ggcgtcgtcc agcatctgct ggatgacgtc gtcctcgggc ttcagcttgg
     2101 ccaggcgggc gcggctgcgc tcggtgatct gctcgtacag ccactcgaag gtggccagca
     2161 gcagttccag cttggtcggg aagtggtgcg actgggcgcc gcggctgacg ccggcggcgc
     2221 ccgggacgtc ggcgatgcgg aagccggcat agcccttctc gcgcaggacg cccagggcgg
     2281 cggcgatcag cttgccctgg gtttccatcg cgcgctcggc ctgggtgcgg cgcttcggcg
     2341 acatgatgac cattcggggt ctctccctgc tagcatcagg gttattgtat catgagcgga
     2401 tacatatgtc aatgtaccgg aacaaacaga caatctggtc tgtttgtaca atattgacgc
     2461 cctgtggggg catccgtatg attgcgcctc aacaaacaga caatctggtc tgtttgtaac
     2521 tagtagagga agcttccgca tggtgagcaa aggtgaagag gataatatgg caagcctgcc
     2581 tgcaacccat gaactgcata tttttggtag cattaacggc gtggattttg atatggttgg
     2641 tcagggcacc ggtaatccga atgatggtta tgaagaactg aatctgaaaa gcaccaaagg
     2701 cgatctgcag tttagcccgt ggattctggt tccgcatatt ggttatggtt ttcatcagta
     2761 tctgccgtat ccggatggta tgagcccgtt tcaggcagca atggttgatg gtagcggtta
     2821 tcaggttcat cgtaccatgc agtttgaaga tggtgcaagc ctgaccgtta attatcgtta
     2881 tacctatgaa ggcagccaca ttaaaggcga agcacaggtt aaaggcaccg gttttccggc
     2941 agatggtccg gttatgacca atagtctgac cgcagcagat tggtgtcgta gcaaaaaaac
     3001 ctatccgaac gataaaacca tcatcagcac cttcaaatgg tcatatacca ccggcaatgg
     3061 taaacgttat cgtagcaccg cacgtaccac ctataccttt gcaaaaccga tggcagcaaa
     3121 ctatctgaaa aatcagccga tgtatgtgtt ccgtaaaacc gaactgaaac acagcaaaac
     3181 agagctgaac tttaaagaat ggcagaaagc ctttaccgat gtgatgggta tggatgagct
     3241 gtataaataa ggtaccttta tagctagctt gacatcccat agaagtgata gagatactga
     3301 gcacaaagaa acacgaggaa aactaaacta gtatgagcaa gaaatatggt gaagccgtga
     3361 tcaaagaatt tatgcgcttt aaagttcaca tggaaggtag catgaatggc cacgaatttg
     3421 aaattgaagg tgaaggcgaa ggtcgtccgt atgaaggcac ccagaccgca aaactgaaag
     3481 ttaccaaagg tggtccgctg ccgtttagct gggatattct gagtccgcag tttatgtatg
     3541 gtagccgtgc atttatcaaa catccggcag atattccgga ttactataaa cagagctttc
     3601 cggaaggttt taaatgggaa cgtgtgatga attttgaaga tggtggtgca gttaccgtta
     3661 cacaggatac cagcctggaa gatggcaccc tgatctataa agttaaactg cgtggcacca
     3721 attttccgcc tgatggtccg gttatgcaga aaaaaaccat gggttgggaa gcaagcaccg
     3781 aacgtctgta tcctgaagat ggcgttctga aaggtgatat caaaatggca ctgcgtctga
     3841 aagatggcgg tcgttatctg gcagatttca aaaccaccta caaagccaaa aaaccggttc
     3901 agatgcctgg tgcatataat gttgatcgca aactggatat taccagccac aatgaagatt
     3961 ataccgtggt ggaacagtat gaacgtagcg aaggccgtca tagtaccggt ggtatggatg
     4021 aactgtacaa gtgagctagt aaaaaagagc tccaattggg atccggacta caaggaccac
     4081 gacggcgact ataaggacca cgacatcgac tacaaggatg acgacgacaa gtgaggatcc
     4141 ggtgaagtga cccgcaaaaa aaaaggccag gggatcgctc ccccggcctt ttttttgaat
     4201 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4261 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4321 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     4381 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4441 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4501 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4561 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4621 ctatttttat aggttaatgt catgataata atggtttctt agcacccttt ctcggtcctt
     4681 caacgttcct gacaacgagc ctccttttcg ccaatccatc gacaatcacc gcgagtccct
     4741 gctcgaacgc tgcgtccgga ccggcttcgt cgaaggcgtc tatcgcggcc cgcaacagcg
     4801 gcgagagcgg agcctgttca acggtgccgc cgcgctcgcc ggcatcgctg tcgccggcct
     4861 gctcctcaag cacggcccca acagtgaagt agctgattgt catcagcgca ttgacggcgt
     4921 ccccggccga aaaacccgcc tcgcagagga agcgaagctg cgcgtcggcc gtttccatct
     4981 gcggtgcgcc cggtcgcgtg ccggcatgga tgcgcgcgcc atcgcggtag gcgagcagcg
     5041 cctgcctgaa gctgcgggca ttcccgatca gaaatgagcg ccagtcgtcg tcggctctcg
     5101 gcaccgaatg cgtatgattc tccgccagca tggcttcggc cagtgcgtcg agcagcgccc
     5161 gcttgttcct gaagtgccag taaagcgccg gctgctgaac ccccaaccgt tccgccagtt
     5221 tgcgtgtcgt cagaccgtct acgccgacct cgttcaacag gtccagggcg gcacggatca
     5281 ctgtattcgg ctgcaacttt gtcatgcttg acactttatc actgataaac ataatatgtc
     5341 caccaactta tcagtgataa agaatccgcg cgttcaatcg gaccagcgga ggctggtccg
     5401 gaggccagac gtgaaaccca acatacccct gatcgtaatt ctgagcactg tcgcgctcga
     5461 cgctgtcggc atcggcctga ttatgccggt gctgccgggc ctcctgcgcg atctggttca
     5521 ctcgaacgac gtcaccgccc actatggcat tctgctggcg ctgtatgcgt tggtgcaatt
     5581 tgcctgcgca cctgtgctgg gcgcgctgtc ggatcgtttc gggcggcggc caatcttgct
     5641 cgtctcgctg gccggcgcca ctgtcgacta cgccatcatg gcgacagcgc ctttcctttg
     5701 ggttctctat atcgggcgga tcgtggccgg catcaccggg gcgactgggg cggtagccgg
     5761 cgcttatatt gccgatatca ctgatggcga tgagcgcgcg cggcacttcg gcttcatgag
     5821 cgcctgtttc gggttcggga tggtcgcggg acctgtgctc ggtgggctga tgggcggttt
     5881 ctccccccac gctccgttct tcgccgcggc agccttgaac ggcctcaatt tcctgacggg
     5941 ctgtttcctt ttgccggagt cgcacaaagg cgaacgccgg ccgttacgcc gggaggctct
     6001 caacccgctc gcttcgttcc ggtgggcccg gggcatgacc gtcgtcgccg ccctgatggc
     6061 ggtcttcttc atcatgcaac ttgtcggaca ggtgccggcc gcgctttggg tcattttcgg
     6121 cgaggatcgc tttcactggg acgcgaccac gatcggcatt tcgcttgccg catttggcat
     6181 tctgcattca ctcgcccagg caatgatcac cggccctgta gccgcccggc tcggcgaaag
     6241 gcgggcactc atgctcggaa tgattgccga cggcacaggc tacatcctgc ttgccttcgc
     6301 gacacgggga tggatggcgt tcccgatcat ggtcctgctt gcttcgggtg gcatcggaat
     6361 gccggcgctg caagcaatgt tgtccaggca ggtggatgag gaacgtcagg ggcagctgca
     6421 aggctcactg gcggcgctca ccagcctgac ctcgatcgtc ggacccctcc tcttcacggc
     6481 gatctatgcg gcttctataa caacgtggaa cgggtgggca tggattgcag gcgctgccct
     6541 ctacttgctc tgcctgccgg cgctgcgtcg cgggctttgg agcggcgcag ggcaacgagc
     6601 cgatcgctga tcgtggaaac gataggccta tgccatgcgg gtcaaggcga cttccggcaa
     6661 gctatacgcg ccctagaatt gtcaatttta atcctctgtt tatcggcagt tcgtagagcg
     6721 cgccgtgcgt cccgagcgat actgagcgaa gcaagtgcgt cgagcagtgc ccgcttgttc
     6781 ctgaaatgcc agtaaagcgc tggctgctga acccccagcc ggaactgacc ccacaaggcc
     6841 ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca ggccgctgcc
     6901 tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc
     6961 cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc ggtgcgagct
     7021 gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct cccatatgaa
     7081 tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca ggacctggca
     7141 acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg acaccgattc
     7201 caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca
     7261 ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct ggcaaagctc
     7321 gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg
     7381 ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac
     7441 gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt
     7501 gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg
     7561 ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct tcgtgtgttt
     7621 cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc aggtcctcgc cggcggtttt
     7681 tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc
     7741 cgcctcctgt tcgagacgac gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg
     7801 gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga ccatcgagcc
     7861 gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga tggtttcggc
     7921 atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt
     7981 ccgattcatt caccctcctt gcgggattgc cccgactcac gccggggcaa tgtgccctta
     8041 ttcctgattt gacccgcctg gtgccttggt gtccagataa tccaccttat cggcaatgaa
     8101 gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg gtattccgaa tcttgccctg
     8161 cacgaatacc agctccgcga agtcgctctt cttgatggag cgcatgggga cgtgcttggc
     8221 aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct gccctcgggc
     8281 ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca
     8341 gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt
     8401 tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct
     8461 catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg taggcctaca
     8521 ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt
     8581 acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
     8641 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
     8701 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     8761 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     8821 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     8881 ccccgaagca gggttatgca gcggaaaaga tccgtcgacc ctttccgacg ctcaccgg
//
