LOCUS       sequence-437185-        5396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EryA sensor with further increased sensitivity by mutation of mphR 
            (T17R), in addition to mphA phosphorylation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5396)
  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 5396)
  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: pCM98
FEATURES             Location/Qualifiers
     source          1..5396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(59..127,128..919)
                     /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     complement(277..296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1090..1678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1579..1598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1767..1784)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1937..2023
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             2260..2973
                     /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"
     RBS             3266..3274
                     /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)"
     terminator      complement(3881..3975)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        complement(5057..5161)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5229..5247
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      5315..5362
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
       61 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1021 caaaatccct taacgtgggt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1081 aggatcatct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1201 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcaggctgt
     1741 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1801 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1861 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1921 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1981 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     2041 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     2101 gcatcaaatt aagcagaagg ccatccgtca ggatggcctt tttgcgtttc tacaaactct
     2161 gctagcaagt aaggccgacg gattgaatat aaccgacgtg actgttacat ttaggtggct
     2221 aaacccgtca acatttaact ttaagaagga gatatacata tgcgtaaagg tgaagaactg
     2281 ttcaccggtg ttgttccgat cctggttgaa ctggacggtg acgttaacgg tcacaaattc
     2341 tctgttcgtg gtgaaggtga aggtgacgct accaacggta aactgaccct gaaattcatc
     2401 tgcaccaccg gtaaactgcc ggttccgtgg ccgaccctgg ttaccaccct gacctacggt
     2461 gttcagtgct tcgctcgtta cccggaccac atgaaacagc acgacttctt caaatctgct
     2521 atgccggaag gttacgttca ggaacgtacc atctctttca aagacgacgg tacctacaaa
     2581 acccgtgctg aagttaaatt cgaaggtgac accctggtta accgtatcga actgaaaggt
     2641 atcgacttca aagaagacgg taacatcctg ggtcacaaac tggaatacaa cttcaactct
     2701 cacaacgttt acatcaccgc tgacaaacag aaaaacggta tcaaagctaa cttcaaaatc
     2761 cgtcacaacg ttgaagacgg ttctgttcag ctggctgacc actaccagca gaacaccccg
     2821 atcggtgacg gtccggttct gctgccggac aaccactacc tgtctaccca gtctgttctg
     2881 tctaaagacc cgaacgaaaa acgtgaccac atggttctgc tggaattcgt taccgctgct
     2941 ggtatcaccc acggtatgga cgaactgtac aaagatgcat gccagttcta gcataaccct
     3001 aatgagtgag ctaacttaca ttaattgcgt tgcgccttaa ttaacggcac tcctcagcaa
     3061 atataatgac cctcttgata acccaagagg gcatttttta atgcccatgg cgtttaccac
     3121 agctaacacc acgtcgtccc tatctgctgc cctaggtcta tgagtggttg ctggataact
     3181 ttacgggcat gcataaggct cgtaggctat attcagggag accacaacgg tttccctcta
     3241 caaataattt tgtttaactt tgaaataagg aggtaataca aatgccgcgt ccgaaactga
     3301 aatctgacga cgaagttctg gaagcggcga gggttgttct gaaacgttgc ggtccgatcg
     3361 aattcaccct gtctggtgtt gcgaaagaag ttggtctgtc tcgtgcggcg ctgatccagc
     3421 gtttcaccaa ccgtgacacc ctgctggttc gtatgatgga acgtggtgtt gaacaggttc
     3481 gtcactacct gaacgcgatc ccgatcggtg cgggtccgca gggtctgtgg gaattcctgc
     3541 aggttctggt tcgttctatg aacacccgta acgacttctc tgttaactac ctgatctctt
     3601 ggtacgaact gcaggttccg gaactgcgta ccctggcgat ccagcgtaac cgtgcggttg
     3661 ttgaaggtat ccgtaaacgt ctgccgccgg gtgcgccggc ggcggcggaa ctgctgctgc
     3721 actctgttat cgcgggtgcg accatgcagt gggcggttga cccggacggt gaactggcgg
     3781 accacgttct ggcgcagatc gcggcgatcc tgtgcctgat gttcccggaa cacgacgact
     3841 tccagctgct gcagccgcac gcgtaataac tagtgcttgg attctcacca ataaaaaacg
     3901 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     3961 ctatcaacag gagtccaagc gagctcgtaa acttggtctg acagccctta acttacttat
     4021 taaaggctat atcgacgttc gctcattccg ctgcggcgag ctgcgccttc gccgcagcga
     4081 ggtactcttc gttacccgag tcgagggcga agagtgcgta ggtgaccgcc ccgaacgcaa
     4141 ggcgctccgc gatgtggtgg gcgagccgcg gccacacccg gccaccggcc gcttcatacg
     4201 tgaggaggag cttcgcgagc ccctcttcac caaagaccat aaggtgcgcg gccatgtcga
     4261 tggcagggtc atcaacgcgg gcctcgctcc agtcgatcat cccgctgacg cgctccgtgt
     4321 tgtcgatgag cacatggccc acgtagagat cgccatgcac caccacggag aaatctggcc
     4381 acgacgaatc gtcgtcgagc cagcgctgcc accggtggag gcgcttgtcg ttcaccacga
     4441 actcgcgtcg gacgcggtca acgtcgtcgg ccaccttctg acgggcctgc gtcggtgtac
     4501 ggatgagcat ccccgcatcc acggcggcgg aaatggggac ggcatgcagg gcggcgagcg
     4561 cggtcgcgaa gctctccgcg aagacctccg agtcctgcgg cacgacccag tcgggcgtgg
     4621 acgaaccagg ctggatgacc atcgcagtcg agtcttcgag catgggatag gcaacgagct
     4681 cggcgttggc cacgcgccag tccggcaccg cgaacggcag gcgattcttg agcattgcca
     4741 gcacccgcgc ctctggttcg accttcgcgc ttacctcggc tcggcgcggg atgcgcagca
     4801 cccaccgacg tccatcgtcg acggtggcga tcacgatcct atagtcgagc ccaagctcat
     4861 tgacagtcag cgggccatgg agcttgagcc catgtcgggc tgcaagtgcg tacagttggg
     4921 aggtatcggc ggtcgtgact acggtcatga ttcactcctg agggcttgac gggtttagcc
     4981 acctaaatgt aacagtcacg ttggttatat tcaatccggc actgttgcga ctgtgtttta
     5041 tatttttctc gttcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5101 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5161 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5221 cctataaaaa taggcgtatc acgaggccct ttcgtcttca ctaagcgact aaaaaattga
     5281 atgtaggaaa ccaacatgcc agttctagca ataactagca taaccccttg gggcctctaa
     5341 acgggtcttg aggggttttt tgctgaaacc tcaggcgaga agcacacggt cacact
//
