LOCUS       sequence-437224-        5395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EryA sensor with increased sensitivity due to phosphorylation of 
            EryA by mphA, which traps EryA in cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5395)
  AUTHORS   Miller CA, Ho JM, Parks SE, Bennett MR
  TITLE     Macrolide Biosensor Optimization through Cellular Substrate 
            Sequestration.
  JOURNAL   ACS Synth Biol. 2021 Feb 19;10(2):258-264. doi: 
            10.1021/acssynbio.0c00572. Epub 2021 Feb 8.
  PUBMED    33555859
REFERENCE   2  (bases 1 to 5395)
  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"; doi: 
            "10.1021/acssynbio.0c00572"; journalName: "ACS Synth Biol"; date: 
            "2021-02-19- 19"; volume: "10"; issue: "2"; pages: "258-264"
COMMENT     Sequence Label: pCM54
FEATURES             Location/Qualifiers
     source          1..5395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(35..82)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(150..168)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        236..340
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1421..1515
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     RBS             2122..2130
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(2423..3136)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      complement(3373..3459)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3612..3629
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     rep_origin      complement(3718..4306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3798..3817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4477..5268,5269..5337))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5100..5119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agtgtgaccg tgtgcttctc gcctgaggtt tcagcaaaaa acccctcaag acccgtttag
       61 aggccccaag gggttatgct agttattgct agaactggca tgttggtttc ctacattcaa
      121 ttttttagtc gcttagtgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      181 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      241 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      301 accctgataa atgcttcaat aatattgaaa aaggaagagt atgaacagaa aaatataaaa
      361 cacagtcgca acagtgccgg attgaatata accgacgtga ctgttacatt taggtggcta
      421 aacccgtcaa gccctcagga gtgaatcatg accgtagtca cgaccgccga tacctcccaa
      481 ctgtacgcac ttgcagcccg acatgggctc aagctccatg gcccgctgac tgtcaatgag
      541 cttgggctcg actataggat cgtgatcgcc accgtcgacg atggacgtcg gtgggtgctg
      601 cgcatcccgc gccgagccga ggtaagcgcg aaggtcgaac cagaggcgcg ggtgctggca
      661 atgctcaaga atcgcctgcc gttcgcggtg ccggactggc gcgtggccaa cgccgagctc
      721 gttgcctatc ccatgctcga agactcgact gcgatggtca tccagcctgg ttcgtccacg
      781 cccgactggg tcgtgccgca ggactcggag gtcttcgcgg agagcttcgc gaccgcgctc
      841 gccgccctgc atgccgtccc catttccgcc gccgtggatg cggggatgct catccgtaca
      901 ccgacgcagg cccgtcagaa ggtggccgac gacgttgacc gcgtccgacg cgagttcgtg
      961 gtgaacgaca agcgcctcca ccggtggcag cgctggctcg acgacgattc gtcgtggcca
     1021 gatttctccg tggtggtgca tggcgatctc tacgtgggcc atgtgctcat cgacaacacg
     1081 gagcgcgtca gcgggatgat cgactggagc gaggcccgcg ttgatgaccc tgccatcgac
     1141 atggccgcgc accttatggt ctttggtgaa gaggggctcg cgaagctcct cctcacgtat
     1201 gaagcggccg gtggccgggt gtggccgcgg ctcgcccacc acatcgcgga gcgccttgcg
     1261 ttcggggcgg tcacctacgc actcttcgcc ctcgactcgg gtaacgaaga gtacctcgct
     1321 gcggcgaagg cgcagctcgc cgcagcggaa tgagcgaacg tcgatatagc ctttaataag
     1381 taagttaagg gctgtcagac caagtttacg agctcgcttg gactcctgtt gatagatcca
     1441 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     1501 tttattggtg agaatccaag cactagttat tacgcgtgcg gctgcagcag ctggaagtcg
     1561 tcgtgttccg ggaacatcag gcacaggatc gccgcgatct gcgccagaac gtggtccgcc
     1621 agttcaccgt ccgggtcaac cgcccactgc atggtcgcac ccgcgataac agagtgcagc
     1681 agcagttccg ccgccgccgg cgcacccggc ggcagacgtt tacggatacc ttcaacaacc
     1741 gcacggttac gctggatcgc cagggtacgc agttccggaa cctgcagttc gtaccaagag
     1801 atcaggtagt taacagagaa gtcgttacgg gtgttcatag aacgaaccag aacctgcagg
     1861 aattcccaca gaccctgcgg acccgcaccg atcgggatcg cgttcaggta gtgacgaacc
     1921 tgttcaacac cacgttccat catacgaacc agcagggtgt cacggttggt gaaacgctgg
     1981 atcagcgccg cacgagacag accaacttct ttcgcaacac cagacagggt gaattcgatc
     2041 ggaccgcaac gtttcagaac aacggtcgcc gcttccagaa cttcgtcgtc agatttcagt
     2101 ttcggacgcg gcatttgtat tacctcctta tttcaaagtt aaacaaaatt atttgtagag
     2161 ggaaaccgtt gtggtctccc tgaatatagc ctacgagcct tatgcatgcc cgtaaagtta
     2221 tccagcaacc actcatagac ctagggcagc agatagggac gacgtggtgt tagctgtggt
     2281 aaacgccatg ggcattaaaa aatgccctct tgggttatca agagggtcat tatatttgct
     2341 gaggagtgcc gttaattaag gcgcaacgca attaatgtaa gttagctcac tcattagggt
     2401 tatgctagaa ctggcatgca tctttgtaca gttcgtccat accgtgggtg ataccagcag
     2461 cggtaacgaa ttccagcaga accatgtggt cacgtttttc gttcgggtct ttagacagaa
     2521 cagactgggt agacaggtag tggttgtccg gcagcagaac cggaccgtca ccgatcgggg
     2581 tgttctgctg gtagtggtca gccagctgaa cagaaccgtc ttcaacgttg tgacggattt
     2641 tgaagttagc tttgataccg tttttctgtt tgtcagcggt gatgtaaacg ttgtgagagt
     2701 tgaagttgta ttccagtttg tgacccagga tgttaccgtc ttctttgaag tcgatacctt
     2761 tcagttcgat acggttaacc agggtgtcac cttcgaattt aacttcagca cgggttttgt
     2821 aggtaccgtc gtctttgaaa gagatggtac gttcctgaac gtaaccttcc ggcatagcag
     2881 atttgaagaa gtcgtgctgt ttcatgtggt ccgggtaacg agcgaagcac tgaacaccgt
     2941 aggtcagggt ggtaaccagg gtcggccacg gaaccggcag tttaccggtg gtgcagatga
     3001 atttcagggt cagtttaccg ttggtagcgt caccttcacc ttcaccacga acagagaatt
     3061 tgtgaccgtt aacgtcaccg tccagttcaa ccaggatcgg aacaacaccg gtgaacagtt
     3121 cttcaccttt acgcatatgt atatctcctt cttaaagtta aatgttgacg ggtttagcca
     3181 cctaaatgta acagtcacgt cggttatatt caatccgtcg gccttacttg ctagcagagt
     3241 ttgtagaaac gcaaaaaggc catcctgacg gatggccttc tgcttaattt gatgcctggc
     3301 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc
     3361 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     3421 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3481 tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     3541 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc
     3601 ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc
     3661 ctgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3721 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3781 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3841 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3901 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3961 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4021 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4081 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4141 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4201 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4261 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagatg atcctttgat
     4321 cttttctacg gggtctgacg ctcagtggaa cgaaaaccca cgttaaggga ttttggtcat
     4381 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4441 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4501 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4561 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4621 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4681 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4741 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4801 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4861 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4921 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4981 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5041 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5101 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5161 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5221 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5281 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5341 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcgga
//
