LOCUS       sequence-437182-        4598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cdGreen expressed in operon with mScarlet-I from an aTc-inducible 
            promoter (Internal Lab ID: UJ11206).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4598)
  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 4598)
  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: p2H12ref
FEATURES             Location/Qualifiers
     source          1..4598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(25..43,44..49,50..66,67..72,73..98)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    25..43
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    50..68
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             124..747
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(197..216)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     1278..1299
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1405..1426)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        2819..2923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2924..2992,2993..3784)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3142..3161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3955..4543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4444..4463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acatgttttc gtcttcacct cgagtcccta tcagtgatag agattgacat ccctatcagt
       61 gatagagata ctgagcacat cagcaggacg cactgaccga atttattaag gaggaggtga
      121 tccatgtcta gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
      181 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
      241 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
      301 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
      361 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
      421 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
      481 ttatgccaac aaggtttttc actagagaat gcattatatg cgctcagcgc tgtggggcat
      541 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
      601 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
      661 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
      721 caacttaaat gtgaaagtgg gtcttaataa taataattcc gaaggaggag gtgatccatg
      781 aattcagagc cgccgccgaa gctcgtcctt gacctggagc gcctcgcgca cgtcccccag
      841 gagaaggcgg gcccgctcca gcgctacgcg gcgacgatcc agtcgcagcg cggcgactac
      901 aacggcaagg tgctgtcgat ccgtcaggac gacctgcgca ccctggccgt gatctacgac
      961 cagtcgccct cggtcctcac cgagcagctg atcagctggg gcgtcctcga cgcggacgcg
     1021 cgccgcgccg tcgcccacga ggagaacccg aagaatatgc cggtcccgta taacgtcttt
     1081 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     1141 gaggacggcg gcgtgcagct cgcctatcac taccagcaga acacccccat cggcgacggc
     1201 cccgtgctgc tgcccgacaa ccactacctg ggcgtgcagt ccaaactgag caaagacccc
     1261 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1321 ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag
     1381 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1441 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1501 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1561 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1621 gccatgcccg aaggctacat tcaggagcgc accatcttct tcaaggacga cggcaactat
     1681 aagacacgcg ctgaggttaa gttcgagggc gacactctgg ttaaccgcat cgagctgaag
     1741 ggcatcgact tcaaggagga cggcaacatc ctgggccata agcttgaata taacatgttg
     1801 tcggagccgg agccgccgcc gaagctcgtc cttgacctgg agcgcctcgc gcacgtcccc
     1861 caggagaagg cgggcccgct ccagcgctac gcggcgacga tccagtcgca gcgcggcgac
     1921 tacaacggca aggtgctgtc gatccgtcag gacgacctgc gcaccctggc cgtgatctac
     1981 gaccagtcgc cctcggtcct caccgagcag ctgatcagct ggggcgtcct cgacgcggac
     2041 gcgcgccgcg ccgtcgccca cgaggagaac tgaactagta tgagcaagaa atatggtgaa
     2101 gccgtgatca aagaatttat gcgctttaaa gttcacatgg aaggtagcat gaatggccac
     2161 gaatttgaaa ttgaaggtga aggcgaaggt cgtccgtatg aaggcaccca gaccgcaaaa
     2221 ctgaaagtta ccaaaggtgg tccgctgccg tttagctggg atattctgag tccgcagttt
     2281 atgtatggta gccgtgcatt tatcaaacat ccggcagata ttccggatta ctataaacag
     2341 agctttccgg aaggttttaa atgggaacgt gtgatgaatt ttgaagatgg tggtgcagtt
     2401 accgttacac aggataccag cctggaagat ggcaccctga tctataaagt taaactgcgt
     2461 ggcaccaatt ttccgcctga tggtccggtt atgcagaaaa aaaccatggg ttgggaagca
     2521 agcaccgaac gtctgtatcc tgaagatggc gttctgaaag gtgatatcaa aatggcactg
     2581 cgtctgaaag atggcggtcg ttatctggca gatttcaaaa ccacctacaa agccaaaaaa
     2641 ccggttcaga tgcctggtgc atataatgtt gatcgcaaac tggatattac cagccacaat
     2701 gaagattata ccgtggtgga acagtatgaa cgtagcgaag gccgtcatag taccggtggt
     2761 atggatgaac tgtacaagtg agctagcgac gtcaggtggc acttttcggg gaaatgtgcg
     2821 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2881 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2941 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3001 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3061 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3121 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3181 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3241 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3301 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3361 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3421 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     3481 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3541 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3601 ctggtttatt gctgataaat ctggagccgg tgagcgtggc tctcgcggta tcattgcagc
     3661 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3721 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3781 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3841 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3901 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4081 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4561 ctttttacgg ttcctggcct tttgctggcc ttttgctc
//
