LOCUS       sequence-437190-        6389 bp    DNA     circular SYN 14-APR-2025
DEFINITION  EryA sensor with increased sensitivity (mphA and mphR (T17R) 
            mutation) and high level resistance (ermC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6389)
  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 6389)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pCM121
FEATURES             Location/Qualifiers
     source          1..6389
                     /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(1134..1202,1203..1994)
                     /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(1352..1371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2165..2753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2654..2673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2842..2859)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3012..3098
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3335..4048
                     /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             4341..4349
                     /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(4956..5050)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        complement(6132..6236)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6304..6322
                     /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 caggcgagaa gcacacggtc acacttccgc
     1081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1141 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1201 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1261 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1321 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1381 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1441 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1501 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1561 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1621 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1681 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1741 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1801 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1861 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1921 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1981 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2101 tcccttaacg tgggttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2161 catcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2761 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcag gctgttttgg
     2821 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     2881 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     2941 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     3001 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     3061 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     3121 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     3181 aaattaagca gaaggccatc cgtcaggatg gcctttttgc gtttctacaa actctgctag
     3241 caagtaaggc cgacggattg aatataaccg acgtgactgt tacatttagg tggctaaacc
     3301 cgtcaacatt taactttaag aaggagatat acatatgcgt aaaggtgaag aactgttcac
     3361 cggtgttgtt ccgatcctgg ttgaactgga cggtgacgtt aacggtcaca aattctctgt
     3421 tcgtggtgaa ggtgaaggtg acgctaccaa cggtaaactg accctgaaat tcatctgcac
     3481 caccggtaaa ctgccggttc cgtggccgac cctggttacc accctgacct acggtgttca
     3541 gtgcttcgct cgttacccgg accacatgaa acagcacgac ttcttcaaat ctgctatgcc
     3601 ggaaggttac gttcaggaac gtaccatctc tttcaaagac gacggtacct acaaaacccg
     3661 tgctgaagtt aaattcgaag gtgacaccct ggttaaccgt atcgaactga aaggtatcga
     3721 cttcaaagaa gacggtaaca tcctgggtca caaactggaa tacaacttca actctcacaa
     3781 cgtttacatc accgctgaca aacagaaaaa cggtatcaaa gctaacttca aaatccgtca
     3841 caacgttgaa gacggttctg ttcagctggc tgaccactac cagcagaaca ccccgatcgg
     3901 tgacggtccg gttctgctgc cggacaacca ctacctgtct acccagtctg ttctgtctaa
     3961 agacccgaac gaaaaacgtg accacatggt tctgctggaa ttcgttaccg ctgctggtat
     4021 cacccacggt atggacgaac tgtacaaaga tgcatgccag ttctagcata accctaatga
     4081 gtgagctaac ttacattaat tgcgttgcgc cttaattaac ggcactcctc agcaaatata
     4141 atgaccctct tgataaccca agagggcatt ttttaatgcc catggcgttt accacagcta
     4201 acaccacgtc gtccctatct gctgccctag gtctatgagt ggttgctgga taactttacg
     4261 ggcatgcata aggctcgtag gctatattca gggagaccac aacggtttcc ctctacaaat
     4321 aattttgttt aactttgaaa taaggaggta atacaaatgc cgcgtccgaa actgaaatct
     4381 gacgacgaag ttctggaagc ggcgagggtt gttctgaaac gttgcggtcc gatcgaattc
     4441 accctgtctg gtgttgcgaa agaagttggt ctgtctcgtg cggcgctgat ccagcgtttc
     4501 accaaccgtg acaccctgct ggttcgtatg atggaacgtg gtgttgaaca ggttcgtcac
     4561 tacctgaacg cgatcccgat cggtgcgggt ccgcagggtc tgtgggaatt cctgcaggtt
     4621 ctggttcgtt ctatgaacac ccgtaacgac ttctctgtta actacctgat ctcttggtac
     4681 gaactgcagg ttccggaact gcgtaccctg gcgatccagc gtaaccgtgc ggttgttgaa
     4741 ggtatccgta aacgtctgcc gccgggtgcg ccggcggcgg cggaactgct gctgcactct
     4801 gttatcgcgg gtgcgaccat gcagtgggcg gttgacccgg acggtgaact ggcggaccac
     4861 gttctggcgc agatcgcggc gatcctgtgc ctgatgttcc cggaacacga cgacttccag
     4921 ctgctgcagc cgcacgcgta ataactagtg cttggattct caccaataaa aaacgcccgg
     4981 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
     5041 aacaggagtc caagcgagct cgtaaacttg gtctgacagc ccttaactta cttattaaag
     5101 gctatatcga cgttcgctca ttccgctgcg gcgagctgcg ccttcgccgc agcgaggtac
     5161 tcttcgttac ccgagtcgag ggcgaagagt gcgtaggtga ccgccccgaa cgcaaggcgc
     5221 tccgcgatgt ggtgggcgag ccgcggccac acccggccac cggccgcttc atacgtgagg
     5281 aggagcttcg cgagcccctc ttcaccaaag accataaggt gcgcggccat gtcgatggca
     5341 gggtcatcaa cgcgggcctc gctccagtcg atcatcccgc tgacgcgctc cgtgttgtcg
     5401 atgagcacat ggcccacgta gagatcgcca tgcaccacca cggagaaatc tggccacgac
     5461 gaatcgtcgt cgagccagcg ctgccaccgg tggaggcgct tgtcgttcac cacgaactcg
     5521 cgtcggacgc ggtcaacgtc gtcggccacc ttctgacggg cctgcgtcgg tgtacggatg
     5581 agcatccccg catccacggc ggcggaaatg gggacggcat gcagggcggc gagcgcggtc
     5641 gcgaagctct ccgcgaagac ctccgagtcc tgcggcacga cccagtcggg cgtggacgaa
     5701 ccaggctgga tgaccatcgc agtcgagtct tcgagcatgg gataggcaac gagctcggcg
     5761 ttggccacgc gccagtccgg caccgcgaac ggcaggcgat tcttgagcat tgccagcacc
     5821 cgcgcctctg gttcgacctt cgcgcttacc tcggctcggc gcgggatgcg cagcacccac
     5881 cgacgtccat cgtcgacggt ggcgatcacg atcctatagt cgagcccaag ctcattgaca
     5941 gtcagcgggc catggagctt gagcccatgt cgggctgcaa gtgcgtacag ttgggaggta
     6001 tcggcggtcg tgactacggt catgattcac tcctgagggc ttgacgggtt tagccaccta
     6061 aatgtaacag tcacgttggt tatattcaat ccggcactgt tgcgactgtg ttttatattt
     6121 ttctcgttca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6181 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6241 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6301 aaaaataggc gtatcacgag gccctttcgt cttcactaag cgactaaaaa attgaatgta
     6361 ggaaaccaac atgccagttc tagcaataa
//
