LOCUS       sequence-417527-        8148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pQFT with mNeonGreen under control of PQJ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8148)
  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 8148)
  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
FEATURES             Location/Qualifiers
     source          1..8148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            123..232
                     /note="incP origin of transfer"
     rep_origin      385..1016
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1479..2067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1968..1987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2129..2740)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    2817..2844
                     /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    2887..2914
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     CDS             3672..3737
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3809..3826)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3809..3825)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3818..3840)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4034..4053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4153..4175
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4213..4231)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             5017..6216
                     /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(6447..7595)
                     /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(join(7867..8148,1..90))
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 cttttcctca atcgctcttc gttcgtctgg aaggcagtac accttgatag gtgggctgcc
       61 cttcctggtt ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt
      121 agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac
      181 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
      241 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
      301 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
      361 ggaaaagatc cgtcgaccct ttccgacgct caccgggctg gttgccctcg ccgctgggct
      421 ggcggccgtc tatggccctg caaacgcgcc agaaacgccg tcgaagccgt gtgcgagaca
      481 ccgcggccgc cggcgttgtg gatacctcgc ggaaaacttg gccctcactg acagatgagg
      541 ggcggacgtt gacacttgag gggccgactc acccggcgcg gcgttgacag atgaggggca
      601 ggctcgattt cggccggcga cgtggagctg gccagcctcg caaatcggcg aaaacgcctg
      661 attttacgcg agtttcccac agatgatgtg gacaagcctg gggataagtg ccctgcggta
      721 ttgacacttg aggggcgcga ctactgacag atgaggggcg cgatccttga cacttgaggg
      781 gcagagtgct gacagatgag gggcgcacct attgacattt gaggggctgt ccacaggcag
      841 aaaatccagc atttgcaagg gtttccgccc gtttttcggc caccgctaac ctgtctttta
      901 acctgctttt aaaccaatat ttataaacct tgtttttaac cagggctgcg ccctgtgcgc
      961 gtgaccgcgc acgccgaagg ggggtgcccc cccttctcga accctcccgg cccgctaacg
     1021 cgggcctccc atccccccag gggctgcgcc cctcggccgc gaacggcctc accccaaaaa
     1081 tggcagccaa gctgaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1141 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1201 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1261 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1321 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1381 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1501 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1561 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1621 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1681 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1741 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1801 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1861 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1921 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1981 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2041 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2101 gccttttgct ggccttttgc tcacatgttc agcgcttgaa cttggcgtag cgctcgcggg
     2161 cgatttccag ggtcgagttg cggacgcgct cgaagcgctc cttgtccttc tgccacagcg
     2221 agcggaccgc caggccgcgg accgagttga agatcagcca caggatgtcc tccgcgtcgt
     2281 cgcggctcag gccgcgcgac accagcacgc ccagccacat atcctcgacg acgaagcggt
     2341 tgcgctcgac ggtgcgctgg atgccctcgc gcagcgccgg gtcgcggtcg gcggccacga
     2401 tcaggtccag cgagatcgag aagtcgtcgt ccaggaagaa ctcggcggcg tcgtccagca
     2461 tctgctggat gacgtcgtcc tcgggcttca gcttggccag gcgggcgcgg ctgcgctcgg
     2521 tgatctgctc gtacagccac tcgaaggtgg ccagcagcag ttccagcttg gtcgggaagt
     2581 ggtgcgactg ggcgccgcgg ctgacgccgg cggcgcccgg gacgtcggcg atgcggaagc
     2641 cggcatagcc cttctcgcgc aggacgccca gggcggcggc gatcagcttg ccctgggttt
     2701 ccatcgcgcg ctcggcctgg gtgcggcgct tcggcgacat gatgaccatt cggggtctct
     2761 ccctgctagc atcagggtta ttgtatcatg agcggataca tatgtcaatg taccggaaca
     2821 aacagacaat ctggtctgtt tgtacaatat tgacgccctg tgggggcatc cgtatgattg
     2881 cgcctcaaca aacagacaat ctggtctgtt tgtaactagt agaggaagct tccgcatggt
     2941 gagcaaaggt gaagaggata atatggcaag cctgcctgca acccatgaac tgcatatttt
     3001 tggtagcatt aacggcgtgg attttgatat ggttggtcag ggcaccggta atccgaatga
     3061 tggttatgaa gaactgaatc tgaaaagcac caaaggcgat ctgcagttta gcccgtggat
     3121 tctggttccg catattggtt atggttttca tcagtatctg ccgtatccgg atggtatgag
     3181 cccgtttcag gcagcaatgg ttgatggtag cggttatcag gttcatcgta ccatgcagtt
     3241 tgaagatggt gcaagcctga ccgttaatta tcgttatacc tatgaaggca gccacattaa
     3301 aggcgaagca caggttaaag gcaccggttt tccggcagat ggtccggtta tgaccaatag
     3361 tctgaccgca gcagattggt gtcgtagcaa aaaaacctat ccgaacgata aaaccatcat
     3421 cagcaccttc aaatggtcat ataccaccgg caatggtaaa cgttatcgta gcaccgcacg
     3481 taccacctat acctttgcaa aaccgatggc agcaaactat ctgaaaaatc agccgatgta
     3541 tgtgttccgt aaaaccgaac tgaaacacag caaaacagag ctgaacttta aagaatggca
     3601 gaaagccttt accgatgtga tgggtatgga tgagctgtat aaataaggta ccgagctcca
     3661 attgggatcc ggactacaag gaccacgacg gcgactataa ggaccacgac atcgactaca
     3721 aggatgacga cgacaagtga ggatccggtg aagtgacccg caaaaaaaaa ggccagggga
     3781 tcgctccccc ggcctttttt ttgaattcac tggccgtcgt tttacaacgt cgtgactggg
     3841 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3901 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3961 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4021 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4081 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4141 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4201 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4261 tttcttagca ccctttctcg gtccttcaac gttcctgaca acgagcctcc ttttcgccaa
     4321 tccatcgaca atcaccgcga gtccctgctc gaacgctgcg tccggaccgg cttcgtcgaa
     4381 ggcgtctatc gcggcccgca acagcggcga gagcggagcc tgttcaacgg tgccgccgcg
     4441 ctcgccggca tcgctgtcgc cggcctgctc ctcaagcacg gccccaacag tgaagtagct
     4501 gattgtcatc agcgcattga cggcgtcccc ggccgaaaaa cccgcctcgc agaggaagcg
     4561 aagctgcgcg tcggccgttt ccatctgcgg tgcgcccggt cgcgtgccgg catggatgcg
     4621 cgcgccatcg cggtaggcga gcagcgcctg cctgaagctg cgggcattcc cgatcagaaa
     4681 tgagcgccag tcgtcgtcgg ctctcggcac cgaatgcgta tgattctccg ccagcatggc
     4741 ttcggccagt gcgtcgagca gcgcccgctt gttcctgaag tgccagtaaa gcgccggctg
     4801 ctgaaccccc aaccgttccg ccagtttgcg tgtcgtcaga ccgtctacgc cgacctcgtt
     4861 caacaggtcc agggcggcac ggatcactgt attcggctgc aactttgtca tgcttgacac
     4921 tttatcactg ataaacataa tatgtccacc aacttatcag tgataaagaa tccgcgcgtt
     4981 caatcggacc agcggaggct ggtccggagg ccagacgtga aacccaacat acccctgatc
     5041 gtaattctga gcactgtcgc gctcgacgct gtcggcatcg gcctgattat gccggtgctg
     5101 ccgggcctcc tgcgcgatct ggttcactcg aacgacgtca ccgcccacta tggcattctg
     5161 ctggcgctgt atgcgttggt gcaatttgcc tgcgcacctg tgctgggcgc gctgtcggat
     5221 cgtttcgggc ggcggccaat cttgctcgtc tcgctggccg gcgccactgt cgactacgcc
     5281 atcatggcga cagcgccttt cctttgggtt ctctatatcg ggcggatcgt ggccggcatc
     5341 accggggcga ctggggcggt agccggcgct tatattgccg atatcactga tggcgatgag
     5401 cgcgcgcggc acttcggctt catgagcgcc tgtttcgggt tcgggatggt cgcgggacct
     5461 gtgctcggtg ggctgatggg cggtttctcc ccccacgctc cgttcttcgc cgcggcagcc
     5521 ttgaacggcc tcaatttcct gacgggctgt ttccttttgc cggagtcgca caaaggcgaa
     5581 cgccggccgt tacgccggga ggctctcaac ccgctcgctt cgttccggtg ggcccggggc
     5641 atgaccgtcg tcgccgccct gatggcggtc ttcttcatca tgcaacttgt cggacaggtg
     5701 ccggccgcgc tttgggtcat tttcggcgag gatcgctttc actgggacgc gaccacgatc
     5761 ggcatttcgc ttgccgcatt tggcattctg cattcactcg cccaggcaat gatcaccggc
     5821 cctgtagccg cccggctcgg cgaaaggcgg gcactcatgc tcggaatgat tgccgacggc
     5881 acaggctaca tcctgcttgc cttcgcgaca cggggatgga tggcgttccc gatcatggtc
     5941 ctgcttgctt cgggtggcat cggaatgccg gcgctgcaag caatgttgtc caggcaggtg
     6001 gatgaggaac gtcaggggca gctgcaaggc tcactggcgg cgctcaccag cctgacctcg
     6061 atcgtcggac ccctcctctt cacggcgatc tatgcggctt ctataacaac gtggaacggg
     6121 tgggcatgga ttgcaggcgc tgccctctac ttgctctgcc tgccggcgct gcgtcgcggg
     6181 ctttggagcg gcgcagggca acgagccgat cgctgatcgt ggaaacgata ggcctatgcc
     6241 atgcgggtca aggcgacttc cggcaagcta tacgcgccct agaattgtca attttaatcc
     6301 tctgtttatc ggcagttcgt agagcgcgcc gtgcgtcccg agcgatactg agcgaagcaa
     6361 gtgcgtcgag cagtgcccgc ttgttcctga aatgccagta aagcgctggc tgctgaaccc
     6421 ccagccggaa ctgaccccac aaggccctag cgtttgcaat gcaccaggtc atcattgacc
     6481 caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc gacctgctcg
     6541 cgccacttct tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt ttccagcttg
     6601 agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc cgtcggcgac
     6661 agcttgcggt acttctccca tatgaatttc gtgtagtggt cgccagcaaa cagcacgacg
     6721 atttcctcgt cgatcaggac ctggcaacgg gacgttttct tgccacggtc caggacgcgg
     6781 aagcggtgca gcagcgacac cgattccagg tgcccaacgc ggtcggacgt gaagcccatc
     6841 gccgtcgcct gtaggcgcga caggcattcc tcggccttcg tgtaataccg gccattgatc
     6901 gaccagccca ggtcctggca aagctcgtag aacgtgaagg tgatcggctc gccgataggg
     6961 gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc ggcccgcagc
     7021 tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac cttgttttgc
     7081 agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg ggccgtgtcg
     7141 tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga aagctgcatt
     7201 tccttgatct gctgcttcgt gtgtttcagc aacgcggcct gcttggcctc gctgacctgt
     7261 tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc tcgcgtgtcg
     7321 atggtcatcg acttcgccaa acctgccgcc tcctgttcga gacgacgcga acgctccacg
     7381 gcggccgatg gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg ctcgatcttg
     7441 gccgtagctt gctggaccat cgagccgacg gactggaagg tttcgcgggg cgcacgcatg
     7501 acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc gatcagttct
     7561 tgcctgtatg ccttccggtc aaacgtccga ttcattcacc ctccttgcgg gattgccccg
     7621 actcacgccg gggcaatgtg cccttattcc tgatttgacc cgcctggtgc cttggtgtcc
     7681 agataatcca ccttatcggc aatgaagtcg gtcccgtaga ccgtctggcc gtccttctcg
     7741 tacttggtat tccgaatctt gccctgcacg aataccagct ccgcgaagtc gctcttcttg
     7801 atggagcgca tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta
     7861 aaaaagtcat ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc
     7921 ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt
     7981 gcgcgggtcg tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt
     8041 gatgcgggcc agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg
     8101 gccgacggcc agcaggtagg cctacaggct catgccggcc gccgccgc
//
