LOCUS       sequence-437178-        9303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cdGreen2 expressed from a cumate-inducible promoter in an operon 
            with mScarlet-I (downstream) as a reference FP (Internal Lab ID: 
            UJ11240).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9303)
  AUTHORS   Kaczmarczyk A, van Vliet S, Jakob RP, Teixeira RD, Scheidat I, 
            Reinders A, Klotz A, Maier T, Jenal U
  TITLE     A genetically encoded biosensor to monitor dynamic changes of 
            c-di-GMP with high temporal resolution.
  JOURNAL   Nat Commun. 2024 May 9;15(1):3920. doi: 10.1038/s41467-024-48295-0.
  PUBMED    38724508
REFERENCE   2  (bases 1 to 9303)
  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: "Nat 
            Commun."; date: "2024-05-9"; volume: "15(1):3920. doi"; pages: " 
            10.1038/s41467-024-48295-0"
COMMENT     Sequence Label: pQFmcs-2H12.D11-scarREF
FEATURES             Location/Qualifiers
     source          1..9303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1594..1613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1755..2366)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    2443..2470
                     /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    2513..2540
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     primer_bind     3068..3089
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3195..3216)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4590..4607)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4590..4606)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4599..4621)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4815..4834)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4934..4956
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4994..5012)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             5798..6997
                     /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(7228..8376)
                     /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(8648..9019)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            9052..9161
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atgttcagcg cttgaacttg gcgtagcgct cgcgggcgat ttccagggtc
     1801 gagttgcgga cgcgctcgaa gcgctccttg tccttctgcc acagcgagcg gaccgccagg
     1861 ccgcggaccg agttgaagat cagccacagg atgtcctccg cgtcgtcgcg gctcaggccg
     1921 cgcgacacca gcacgcccag ccacatatcc tcgacgacga agcggttgcg ctcgacggtg
     1981 cgctggatgc cctcgcgcag cgccgggtcg cggtcggcgg ccacgatcag gtccagcgag
     2041 atcgagaagt cgtcgtccag gaagaactcg gcggcgtcgt ccagcatctg ctggatgacg
     2101 tcgtcctcgg gcttcagctt ggccaggcgg gcgcggctgc gctcggtgat ctgctcgtac
     2161 agccactcga aggtggccag cagcagttcc agcttggtcg ggaagtggtg cgactgggcg
     2221 ccgcggctga cgccggcggc gcccgggacg tcggcgatgc ggaagccggc atagcccttc
     2281 tcgcgcagga cgcccagggc ggcggcgatc agcttgccct gggtttccat cgcgcgctcg
     2341 gcctgggtgc ggcgcttcgg cgacatgatg accattcggg gtctctccct gctagcatca
     2401 gggttattgt atcatgagcg gatacatatg tcaatgtacc ggaacaaaca gacaatctgg
     2461 tctgtttgta catggttgac ggatcgccgc gaagccgcta actgcgccgg ttaacaaaca
     2521 gacaatctgg tctgtttgta actaggttcc gaaggaggag gtgatccatg aattcagagc
     2581 cgccgccgaa gctcgtcctt gacctggagc gcctcgcgca cgtcccccag gagaaggcgg
     2641 gcccgctcca gcgctacgcg gcgacgatcc agtcgcagcg cggcgactac aacggcaagg
     2701 tgctgtcgat ccgtccggcg gacctgcgca ccctggccgt gatctacgac cagtcgccct
     2761 cggtcctcac cgagcagctg atcagctggg gcgtcctcga cgcggacgcg cgccgcgccg
     2821 tcgcccacga ggagaacccg aagaatatgc cggtcccgta taacgtcttt atcatggccg
     2881 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggcg
     2941 gcgtgcagct cgcctatcac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3001 tgcccgacaa ccactacctg ggcgtgcagt ccaaactgag caaagacccc aacgagaagc
     3061 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3121 agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag ctgttcaccg
     3181 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3241 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3301 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3361 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3421 aaggctacat tcaggagcgc accatcttct tcaaggacga cggcaactat aagacacgcg
     3481 ctgaggttaa gttcgagggc gacactctgg ttaaccgcat cgagctgaag ggcatcgact
     3541 tcaaggagga cggcaacatc ctgggccata agcttgaata taacatgttg tcggagccgg
     3601 agccgccgcc gaagctcgtc cttgacctgg agcgcctcgc gcacgtcccc caggagaagg
     3661 cgggcccgct ccagcgctac gcggcgacga tccagtcgca gcgcggcgac tacaacggca
     3721 aggtgctgtc gatccgtccg gcggacctgc gcaccctggc cgtgatctac gaccagtcgc
     3781 cctcggtcct caccgagcag ctgatcagct ggggcgtcct cgacgcggac gcgcgccgcg
     3841 ccgtcgccca cgaggagaac tgaactagta tgagcaagaa atatggtgaa gccgtgatca
     3901 aagaatttat gcgctttaaa gttcacatgg aaggtagcat gaatggccac gaatttgaaa
     3961 ttgaaggtga aggcgaaggt cgtccgtatg aaggcaccca gaccgcaaaa ctgaaagtta
     4021 ccaaaggtgg tccgctgccg tttagctggg atattctgag tccgcagttt atgtatggta
     4081 gccgtgcatt tatcaaacat ccggcagata ttccggatta ctataaacag agctttccgg
     4141 aaggttttaa atgggaacgt gtgatgaatt ttgaagatgg tggtgcagtt accgttacac
     4201 aggataccag cctggaagat ggcaccctga tctataaagt taaactgcgt ggcaccaatt
     4261 ttccgcctga tggtccggtt atgcagaaaa aaaccatggg ttgggaagca agcaccgaac
     4321 gtctgtatcc tgaagatggc gttctgaaag gtgatatcaa aatggcactg cgtctgaaag
     4381 atggcggtcg ttatctggca gatttcaaaa ccacctacaa agccaaaaaa ccggttcaga
     4441 tgcctggtgc atataatgtt gatcgcaaac tggatattac cagccacaat gaagattata
     4501 ccgtggtgga acagtatgaa cgtagcgaag gccgtcatag taccggtggt atggatgaac
     4561 tgtacaagtg agctagtgac gtcaaattca ctggccgtcg ttttacaacg tcgtgactgg
     4621 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4681 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4741 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4801 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4861 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4921 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4981 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5041 gtttcttagc accctttctc ggtccttcaa cgttcctgac aacgagcctc cttttcgcca
     5101 atccatcgac aatcaccgcg agtccctgct cgaacgctgc gtccggaccg gcttcgtcga
     5161 aggcgtctat cgcggcccgc aacagcggcg agagcggagc ctgttcaacg gtgccgccgc
     5221 gctcgccggc atcgctgtcg ccggcctgct cctcaagcac ggccccaaca gtgaagtagc
     5281 tgattgtcat cagcgcattg acggcgtccc cggccgaaaa acccgcctcg cagaggaagc
     5341 gaagctgcgc gtcggccgtt tccatctgcg gtgcgcccgg tcgcgtgccg gcatggatgc
     5401 gcgcgccatc gcggtaggcg agcagcgcct gcctgaagct gcgggcattc ccgatcagaa
     5461 atgagcgcca gtcgtcgtcg gctctcggca ccgaatgcgt atgattctcc gccagcatgg
     5521 cttcggccag tgcgtcgagc agcgcccgct tgttcctgaa gtgccagtaa agcgccggct
     5581 gctgaacccc caaccgttcc gccagtttgc gtgtcgtcag accgtctacg ccgacctcgt
     5641 tcaacaggtc cagggcggca cggatcactg tattcggctg caactttgtc atgcttgaca
     5701 ctttatcact gataaacata atatgtccac caacttatca gtgataaaga atccgcgcgt
     5761 tcaatcggac cagcggaggc tggtccggag gccagacgtg aaacccaaca tacccctgat
     5821 cgtaattctg agcactgtcg cgctcgacgc tgtcggcatc ggcctgatta tgccggtgct
     5881 gccgggcctc ctgcgcgatc tggttcactc gaacgacgtc accgcccact atggcattct
     5941 gctggcgctg tatgcgttgg tgcaatttgc ctgcgcacct gtgctgggcg cgctgtcgga
     6001 tcgtttcggg cggcggccaa tcttgctcgt ctcgctggcc ggcgccactg tcgactacgc
     6061 catcatggcg acagcgcctt tcctttgggt tctctatatc gggcggatcg tggccggcat
     6121 caccggggcg actggggcgg tagccggcgc ttatattgcc gatatcactg atggcgatga
     6181 gcgcgcgcgg cacttcggct tcatgagcgc ctgtttcggg ttcgggatgg tcgcgggacc
     6241 tgtgctcggt gggctgatgg gcggtttctc cccccacgct ccgttcttcg ccgcggcagc
     6301 cttgaacggc ctcaatttcc tgacgggctg tttccttttg ccggagtcgc acaaaggcga
     6361 acgccggccg ttacgccggg aggctctcaa cccgctcgct tcgttccggt gggcccgggg
     6421 catgaccgtc gtcgccgccc tgatggcggt cttcttcatc atgcaacttg tcggacaggt
     6481 gccggccgcg ctttgggtca ttttcggcga ggatcgcttt cactgggacg cgaccacgat
     6541 cggcatttcg cttgccgcat ttggcattct gcattcactc gcccaggcaa tgatcaccgg
     6601 ccctgtagcc gcccggctcg gcgaaaggcg ggcactcatg ctcggaatga ttgccgacgg
     6661 cacaggctac atcctgcttg ccttcgcgac acggggatgg atggcgttcc cgatcatggt
     6721 cctgcttgct tcgggtggca tcggaatgcc ggcgctgcaa gcaatgttgt ccaggcaggt
     6781 ggatgaggaa cgtcaggggc agctgcaagg ctcactggcg gcgctcacca gcctgacctc
     6841 gatcgtcgga cccctcctct tcacggcgat ctatgcggct tctataacaa cgtggaacgg
     6901 gtgggcatgg attgcaggcg ctgccctcta cttgctctgc ctgccggcgc tgcgtcgcgg
     6961 gctttggagc ggcgcagggc aacgagccga tcgctgatcg tggaaacgat aggcctatgc
     7021 catgcgggtc aaggcgactt ccggcaagct atacgcgccc tagaattgtc aattttaatc
     7081 ctctgtttat cggcagttcg tagagcgcgc cgtgcgtccc gagcgatact gagcgaagca
     7141 agtgcgtcga gcagtgcccg cttgttcctg aaatgccagt aaagcgctgg ctgctgaacc
     7201 cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac
     7261 ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc
     7321 gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt
     7381 gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga
     7441 cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac
     7501 gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg
     7561 gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat
     7621 cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat
     7681 cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg
     7741 ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag
     7801 ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg
     7861 cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc
     7921 gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat
     7981 ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct cgctgacctg
     8041 ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc
     8101 gatggtcatc gacttcgcca aacctgccgc ctcctgttcg agacgacgcg aacgctccac
     8161 ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt
     8221 ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg gcgcacgcat
     8281 gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc
     8341 ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc
     8401 gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg ccttggtgtc
     8461 cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc cgtccttctc
     8521 gtacttggta ttccgaatct tgccctgcac gaataccagc tccgcgaagt cgctcttctt
     8581 gatggagcgc atggggacgt gcttggcaat cacgcgcacc ccccggccgt tttagcggct
     8641 aaaaaagtca tggctctgcc ctcgggcgga ccacgcccat catgaccttg ccaagctcgt
     8701 cctgcttctc ttcgatcttc gccagcaggg cgaggatcgt ggcatcaccg aaccgcgccg
     8761 tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc caggcggccc aggtcgccat
     8821 tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt ttgtagccct
     8881 ggccgacggc cagcaggtag gcctacaggc tcatgccggc cgccgccgcc ttttcctcaa
     8941 tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg tgggctgccc ttcctggttg
     9001 gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc
     9061 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
     9121 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     9181 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     9241 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaagatcc
     9301 gtc
//
