LOCUS       sequence-445574-        8595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cdGreen2 expressed from a cumate-inducible promoter (Internal Lab 
            ID: UJ11216).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8595)
  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 8595)
  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
FEATURES             Location/Qualifiers
     source          1..8595
                     /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(3882..3899)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3882..3898)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3891..3913)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4107..4126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4226..4248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4286..4304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             5090..6289
                     /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(6520..7668)
                     /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(7940..8311)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            8344..8453
                     /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 tgaactagtg acgtcaaatt cactggccgt cgttttacaa
     3901 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     3961 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4021 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4081 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4141 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4201 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4261 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4321 atgataataa tggtttctta gcaccctttc tcggtccttc aacgttcctg acaacgagcc
     4381 tccttttcgc caatccatcg acaatcaccg cgagtccctg ctcgaacgct gcgtccggac
     4441 cggcttcgtc gaaggcgtct atcgcggccc gcaacagcgg cgagagcgga gcctgttcaa
     4501 cggtgccgcc gcgctcgccg gcatcgctgt cgccggcctg ctcctcaagc acggccccaa
     4561 cagtgaagta gctgattgtc atcagcgcat tgacggcgtc cccggccgaa aaacccgcct
     4621 cgcagaggaa gcgaagctgc gcgtcggccg tttccatctg cggtgcgccc ggtcgcgtgc
     4681 cggcatggat gcgcgcgcca tcgcggtagg cgagcagcgc ctgcctgaag ctgcgggcat
     4741 tcccgatcag aaatgagcgc cagtcgtcgt cggctctcgg caccgaatgc gtatgattct
     4801 ccgccagcat ggcttcggcc agtgcgtcga gcagcgcccg cttgttcctg aagtgccagt
     4861 aaagcgccgg ctgctgaacc cccaaccgtt ccgccagttt gcgtgtcgtc agaccgtcta
     4921 cgccgacctc gttcaacagg tccagggcgg cacggatcac tgtattcggc tgcaactttg
     4981 tcatgcttga cactttatca ctgataaaca taatatgtcc accaacttat cagtgataaa
     5041 gaatccgcgc gttcaatcgg accagcggag gctggtccgg aggccagacg tgaaacccaa
     5101 catacccctg atcgtaattc tgagcactgt cgcgctcgac gctgtcggca tcggcctgat
     5161 tatgccggtg ctgccgggcc tcctgcgcga tctggttcac tcgaacgacg tcaccgccca
     5221 ctatggcatt ctgctggcgc tgtatgcgtt ggtgcaattt gcctgcgcac ctgtgctggg
     5281 cgcgctgtcg gatcgtttcg ggcggcggcc aatcttgctc gtctcgctgg ccggcgccac
     5341 tgtcgactac gccatcatgg cgacagcgcc tttcctttgg gttctctata tcgggcggat
     5401 cgtggccggc atcaccgggg cgactggggc ggtagccggc gcttatattg ccgatatcac
     5461 tgatggcgat gagcgcgcgc ggcacttcgg cttcatgagc gcctgtttcg ggttcgggat
     5521 ggtcgcggga cctgtgctcg gtgggctgat gggcggtttc tccccccacg ctccgttctt
     5581 cgccgcggca gccttgaacg gcctcaattt cctgacgggc tgtttccttt tgccggagtc
     5641 gcacaaaggc gaacgccggc cgttacgccg ggaggctctc aacccgctcg cttcgttccg
     5701 gtgggcccgg ggcatgaccg tcgtcgccgc cctgatggcg gtcttcttca tcatgcaact
     5761 tgtcggacag gtgccggccg cgctttgggt cattttcggc gaggatcgct ttcactggga
     5821 cgcgaccacg atcggcattt cgcttgccgc atttggcatt ctgcattcac tcgcccaggc
     5881 aatgatcacc ggccctgtag ccgcccggct cggcgaaagg cgggcactca tgctcggaat
     5941 gattgccgac ggcacaggct acatcctgct tgccttcgcg acacggggat ggatggcgtt
     6001 cccgatcatg gtcctgcttg cttcgggtgg catcggaatg ccggcgctgc aagcaatgtt
     6061 gtccaggcag gtggatgagg aacgtcaggg gcagctgcaa ggctcactgg cggcgctcac
     6121 cagcctgacc tcgatcgtcg gacccctcct cttcacggcg atctatgcgg cttctataac
     6181 aacgtggaac gggtgggcat ggattgcagg cgctgccctc tacttgctct gcctgccggc
     6241 gctgcgtcgc gggctttgga gcggcgcagg gcaacgagcc gatcgctgat cgtggaaacg
     6301 ataggcctat gccatgcggg tcaaggcgac ttccggcaag ctatacgcgc cctagaattg
     6361 tcaattttaa tcctctgttt atcggcagtt cgtagagcgc gccgtgcgtc ccgagcgata
     6421 ctgagcgaag caagtgcgtc gagcagtgcc cgcttgttcc tgaaatgcca gtaaagcgct
     6481 ggctgctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag
     6541 gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc
     6601 gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa
     6661 ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg
     6721 ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc
     6781 aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg
     6841 gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga
     6901 cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata
     6961 ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg
     7021 ctcgccgata ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc
     7081 gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat
     7141 gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga
     7201 gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa
     7261 ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc
     7321 ctcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt
     7381 tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagacgacg
     7441 cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc
     7501 gcgctcgatc ttggccgtag cttgctggac catcgagccg acggactgga aggtttcgcg
     7561 gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc
     7621 gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg
     7681 cgggattgcc ccgactcacg ccggggcaat gtgcccttat tcctgatttg acccgcctgg
     7741 tgccttggtg tccagataat ccaccttatc ggcaatgaag tcggtcccgt agaccgtctg
     7801 gccgtccttc tcgtacttgg tattccgaat cttgccctgc acgaatacca gctccgcgaa
     7861 gtcgctcttc ttgatggagc gcatggggac gtgcttggca atcacgcgca ccccccggcc
     7921 gttttagcgg ctaaaaaagt catggctctg ccctcgggcg gaccacgccc atcatgacct
     7981 tgccaagctc gtcctgcttc tcttcgatct tcgccagcag ggcgaggatc gtggcatcac
     8041 cgaaccgcgc cgtgcgcggg tcgtcggtga gccagagttt cagcaggccg cccaggcggc
     8101 ccaggtcgcc attgatgcgg gccagctcgc ggacgtgctc atagtccacg acgcccgtga
     8161 ttttgtagcc ctggccgacg gccagcaggt aggcctacag gctcatgccg gccgccgccg
     8221 ccttttcctc aatcgctctt cgttcgtctg gaaggcagta caccttgata ggtgggctgc
     8281 ccttcctggt tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg
     8341 tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga
     8401 cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac
     8461 gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg
     8521 cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag
     8581 cggaaaagat ccgtc
//
