LOCUS       sequence-437223-        6388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EryA sensor with increased sensitivity by mphA and high level 
            resistance by ermC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6388)
  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 6388)
  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: pCM120
FEATURES             Location/Qualifiers
     source          1..6388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..48
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      complement(49..143)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        complement(916..1020)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1133..1201,1202..1993)
                     /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(1351..1370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2164..2752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2653..2672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2841..2858)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3011..3097
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3334..4047
                     /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             4340..4348
                     /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(4955..5049)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        complement(6131..6235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6303..6321
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgat tctcaccaat
       61 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
      121 tactggatct atcaacagga gtccaagcga gctcgtaaac ttggtctgac agcccttaac
      181 ttacttatta aataatttat agctattgaa aagagataag aattgttcaa agctaatatt
      241 gtttaaatcg tcaattcctg catgttttaa ggaattgtta aattgatttt ttgtaaatat
      301 tttcttgtat tctttgttaa cccatttcat aacgaaataa ttatactttt gtttatcttt
      361 gtgtgatatt cttgattttt ttctacttaa tctgataagt gagctattca ctttaggttt
      421 aggatgaaaa tattctcttg gaaccatact taatatagaa atatcaactt ctgccattaa
      481 aagtaatgcc aatgagcgtt ttgtatttaa taatctttta gcaaacccgt attccacgat
      541 taaataaatc tcattagcta tactatcaaa aacaattttg cgtattatat ccgtacttat
      601 gttataaggt atattaccat atattttata ggattggttt ttaggaaatt taaactgcaa
      661 tatatccttg tttaaaactt ggaaattatc gtgatcaaca agtttatttt ctgtagtttt
      721 gcataattta tggtctattt caatggcagt tacgaaatta cacctcttta ctaattcaag
      781 ggtaaaatgg ccttttcctg agccgatttc aaagatatta tcatgttcat ttaatcttat
      841 atttgtcatt attttatcta tattatgttt tgaagtaata aagttttgac tgtgttttat
      901 atttttctcg ttcatactct tcctttttca atattattga agcatttatc agggttattg
      961 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1021 cacatttccc cgaaaagtgc cactgaacct aggcgagaag cacacggtca cacttccgct
     1081 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1621 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1681 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1741 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1801 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1861 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1921 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1981 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2041 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2101 cccttaacgt gggttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2161 atcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2221 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2281 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2341 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2401 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2461 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2521 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2581 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2641 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2701 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2761 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcagg ctgttttggc
     2821 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     2881 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     2941 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     3001 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     3061 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     3121 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     3181 aattaagcag aaggccatcc gtcaggatgg cctttttgcg tttctacaaa ctctgctagc
     3241 aagtaaggcc gacggattga atataaccga cgtgactgtt acatttaggt ggctaaaccc
     3301 gtcaacattt aactttaaga aggagatata catatgcgta aaggtgaaga actgttcacc
     3361 ggtgttgttc cgatcctggt tgaactggac ggtgacgtta acggtcacaa attctctgtt
     3421 cgtggtgaag gtgaaggtga cgctaccaac ggtaaactga ccctgaaatt catctgcacc
     3481 accggtaaac tgccggttcc gtggccgacc ctggttacca ccctgaccta cggtgttcag
     3541 tgcttcgctc gttacccgga ccacatgaaa cagcacgact tcttcaaatc tgctatgccg
     3601 gaaggttacg ttcaggaacg taccatctct ttcaaagacg acggtaccta caaaacccgt
     3661 gctgaagtta aattcgaagg tgacaccctg gttaaccgta tcgaactgaa aggtatcgac
     3721 ttcaaagaag acggtaacat cctgggtcac aaactggaat acaacttcaa ctctcacaac
     3781 gtttacatca ccgctgacaa acagaaaaac ggtatcaaag ctaacttcaa aatccgtcac
     3841 aacgttgaag acggttctgt tcagctggct gaccactacc agcagaacac cccgatcggt
     3901 gacggtccgg ttctgctgcc ggacaaccac tacctgtcta cccagtctgt tctgtctaaa
     3961 gacccgaacg aaaaacgtga ccacatggtt ctgctggaat tcgttaccgc tgctggtatc
     4021 acccacggta tggacgaact gtacaaagat gcatgccagt tctagcataa ccctaatgag
     4081 tgagctaact tacattaatt gcgttgcgcc ttaattaacg gcactcctca gcaaatataa
     4141 tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgttta ccacagctaa
     4201 caccacgtcg tccctatctg ctgccctagg tctatgagtg gttgctggat aactttacgg
     4261 gcatgcataa ggctcgtagg ctatattcag ggagaccaca acggtttccc tctacaaata
     4321 attttgttta actttgaaat aaggaggtaa tacaaatgcc gcgtccgaaa ctgaaatctg
     4381 acgacgaagt tctggaagcg gcgaccgttg ttctgaaacg ttgcggtccg atcgaattca
     4441 ccctgtctgg tgttgcgaaa gaagttggtc tgtctcgtgc ggcgctgatc cagcgtttca
     4501 ccaaccgtga caccctgctg gttcgtatga tggaacgtgg tgttgaacag gttcgtcact
     4561 acctgaacgc gatcccgatc ggtgcgggtc cgcagggtct gtgggaattc ctgcaggttc
     4621 tggttcgttc tatgaacacc cgtaacgact tctctgttaa ctacctgatc tcttggtacg
     4681 aactgcaggt tccggaactg cgtaccctgg cgatccagcg taaccgtgcg gttgttgaag
     4741 gtatccgtaa acgtctgccg ccgggtgcgc cggcggcggc ggaactgctg ctgcactctg
     4801 ttatcgcggg tgcgaccatg cagtgggcgg ttgacccgga cggtgaactg gcggaccacg
     4861 ttctggcgca gatcgcggcg atcctgtgcc tgatgttccc ggaacacgac gacttccagc
     4921 tgctgcagcc gcacgcgtaa taactagtgc ttggattctc accaataaaa aacgcccggc
     4981 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     5041 acaggagtcc aagcgagctc gtaaacttgg tctgacagcc cttaacttac ttattaaagg
     5101 ctatatcgac gttcgctcat tccgctgcgg cgagctgcgc cttcgccgca gcgaggtact
     5161 cttcgttacc cgagtcgagg gcgaagagtg cgtaggtgac cgccccgaac gcaaggcgct
     5221 ccgcgatgtg gtgggcgagc cgcggccaca cccggccacc ggccgcttca tacgtgagga
     5281 ggagcttcgc gagcccctct tcaccaaaga ccataaggtg cgcggccatg tcgatggcag
     5341 ggtcatcaac gcgggcctcg ctccagtcga tcatcccgct gacgcgctcc gtgttgtcga
     5401 tgagcacatg gcccacgtag agatcgccat gcaccaccac ggagaaatct ggccacgacg
     5461 aatcgtcgtc gagccagcgc tgccaccggt ggaggcgctt gtcgttcacc acgaactcgc
     5521 gtcggacgcg gtcaacgtcg tcggccacct tctgacgggc ctgcgtcggt gtacggatga
     5581 gcatccccgc atccacggcg gcggaaatgg ggacggcatg cagggcggcg agcgcggtcg
     5641 cgaagctctc cgcgaagacc tccgagtcct gcggcacgac ccagtcgggc gtggacgaac
     5701 caggctggat gaccatcgca gtcgagtctt cgagcatggg ataggcaacg agctcggcgt
     5761 tggccacgcg ccagtccggc accgcgaacg gcaggcgatt cttgagcatt gccagcaccc
     5821 gcgcctctgg ttcgaccttc gcgcttacct cggctcggcg cgggatgcgc agcacccacc
     5881 gacgtccatc gtcgacggtg gcgatcacga tcctatagtc gagcccaagc tcattgacag
     5941 tcagcgggcc atggagcttg agcccatgtc gggctgcaag tgcgtacagt tgggaggtat
     6001 cggcggtcgt gactacggtc atgattcact cctgagggct tgacgggttt agccacctaa
     6061 atgtaacagt cacgttggtt atattcaatc cggcactgtt gcgactgtgt tttatatttt
     6121 tctcgttcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6181 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6241 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6301 aaaataggcg tatcacgagg ccctttcgtc ttcactaagc gactaaaaaa ttgaatgtag
     6361 gaaaccaaca tgccagttct agcaataa
//
