LOCUS       sequence-456014-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  c-di-AMP biosensor plasmid. Encodes yfp downstream of 
            c-di-AMP-binding kimA riboswitch. Plasmid confers ampicillin 
            resistance to E. coli and erythromycin resistance to S. aureus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  AUTHORS   Kviatkovski I, Zhong Q, Vaidya S, Grundling A
  TITLE     Identification of novel genetic factors that regulate c-di-AMP 
            production in Staphylococcus aureus using a riboswitch-based 
            biosensor.
  JOURNAL   mSphere. 2024 Sep 17:e0032124. doi: 10.1128/msphere.00321-24.
  PUBMED    39287429
REFERENCE   2  (bases 1 to 6939)
  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: "mSphere. 
            2024 Sep 17:e0032124. doi: 10.1128/msphere.00321-24."
COMMENT     Sequence Label: pCN34e-kimA-yfp
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3404..3426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3418..3435
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3419..3435
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4867..5455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4947..4966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5626..6417,6418..6486))
                     /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     6249..6268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6487..6591)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6659..6677
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6715..6737)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tatgcatccc ttaacttact tattaaataa tttatagcta ttgaaaagag ataagaattg
       61 ttcaaagcta atattgttta aatcgtcaat tcctgcatgt tttaaggaat tgttaaattg
      121 attttttgta aatattttct tgtattcttt gttaacccat ttcataacga aataattata
      181 cttttgttta tctttgtgtg atattcttga tttttttcta cttaatctga taagtgagct
      241 attcacttta ggtttaggat gaaaatattc tcttggaacc atacttaata tagaaatatc
      301 aacttctgcc attaaaagta atgccaatga gcgttttgta tttaataatc ttttagcaaa
      361 cccgtattcc acgattaaat aaatctcatt agctatacta tcaaaaacaa ttttgcgtat
      421 tatatccgta cttatgttat aaggtatatt accatatatt ttataggatt ggtttttagg
      481 aaatttaaac tgcaatatat ccttgtttaa aacttggaaa ttatcgtgat caacaagttt
      541 attttctgta gttttgcata atttatggtc tatttcaatg gcagttacga aattacacct
      601 ctttactaat tcaagggtaa aatggccttt tcctgagccg atttcaaaga tattatcatg
      661 ttcatttaat cttatatttg tcattatttt atctatatta tgttttgaag taataaagtt
      721 ttgactgtgt tttatatttt tctcgttcat tataaccctc tttaatttgg ttatatgaat
      781 tttgcttatt aacgattcat tataaccact tattttttgt ttggttgata atgaactgtg
      841 ctgattacaa aaatactaaa aatgcccata ttttttcctc cttataaaat tagtataatt
      901 atagcacgag ctctgataaa tatgaacatg atgagtgatc gttaaattta tactgcaatc
      961 ggatgcgatt attgaataaa agatatgaga gatttatcta atttcttttt tcttgtaaaa
     1021 aaagaaagtt cttaaaggtt ttatagtttt ggtcgtagag cacacggttt aacgacttaa
     1081 ttacgaagta aataagtcta gtgtgttaga ctttatgaaa tctatatacg tttatatata
     1141 tttattatcc ggaggtgtag catgtctcat tcaattccta ggtgggccca ataaaagcaa
     1201 tcaatgaacc aagacagctc aaactaatac caataattaa caattttttt atatttatat
     1261 aatcagataa ttttccatat actgctgttc ctatcgaaaa agttaacata tatgcagtgt
     1321 ttacccagtt tgtaattcca ggagtagtat taaaatgatt tgcaatatca ggtaaagaaa
     1381 catttaaaac catttcattt aatacactaa aaaatgaaag aatacaaagc caaaataaaa
     1441 cgctataaaa caaaccttta aattttttat ataaactaaa caattttacc tctttacttg
     1501 aggtgactaa agtttatagg ggtaaattat tctttcacct catataaatt ccccaaattt
     1561 ttaaatagac acttcataaa aaaatcctcc taaattagaa tagtttaaat atatcatttt
     1621 tgttcagtaa tattaatatg tacactattt ccaaaattta aattcatgtt gccaaaaatc
     1681 gatttgtttt tgcaattgtt tttcgttcgc ttttaaagtc gatttcatta agtccgttaa
     1741 atcgactggc gaaatttctt ttatcaaatt cttatatttt gttctagaat ttctgtgaag
     1801 ctttccccat tcttcttcat cactcaataa cataaaaact attgctctat ccgcagtgcg
     1861 ttggatagtt ttccaatctg gttgcaagat atgtaaatca ctaaagcaat cattccagta
     1921 atccaccata tctcttttaa gttcgatttc tacacgccat aaatgttcag acataacttc
     1981 agcatctgca ttatctttac gttcttgctt tttattataa attctaataa atctattact
     2041 atctctcacg ccaaaatatt ttgtttctgg cttaccatta cgaccataaa aaatagtttt
     2101 cttaactgct ttatcagaca ttgcatagta gtcactcaaa tcatcttcaa aatcaaaggc
     2161 taaatctaat cttgtaaaac cgtcatcttc catgtagctt attatatttt gttttaacca
     2221 aatcatttca tctcgtgtaa gtttgtttgg attaaattca atacgcatat tacgtctatc
     2281 ccaactatct gctttcactt tgtcatattc aatataaact ttttcttgaa gtgctttagc
     2341 tttaaacttt gtttgaagaa tatcccaaag tcttatttgg ggctctacac tcataaattt
     2401 agaaagggct tgagcgttgt ctcggttgag atttccaacg atcgtcattg catcaaaact
     2461 taattttgga tcagatatgc acacagtatg tgcgtccaac cggctattag agtagccggt
     2521 tttagaaaaa ttgtctaaat cgtgattttc caaatgattt gaatgatttg catgattgtt
     2581 tttatacata aaaaatcgac tccttaatct caatttcgtt taaggaatcg ctcacccaaa
     2641 tatatatctt gatgtatatt taaatatcgt ttaatatcta aatatacaag attataaaaa
     2701 caactcagtg tttttttctt tgaatgatgt cgttcacaaa ctttggtcag ggcgtgagcg
     2761 actccttttt attttgttat taatataaca ctatcaaaag atttggtcta atcagatcaa
     2821 gtcttttttt atttaagcat ttgtattatc tggtaaacag ttaaaactac taaaacacca
     2881 agtacatact tacttgttat aaaattctca aaggcttctc tgaatttcat ttcttcaact
     2941 ccttttattg ctttattcca atttccttat tggtcggaac ctacagggct ttatgtgttg
     3001 ttgagttggt actttcttgg gattaatccc aattcaagtc caaccaactc gctaacaagt
     3061 tagctaacac atagcccatt ccaaccaata agttttctcg gcataaatgc atgttctata
     3121 agtgcatacc tctatctcgt ttttgattct tttgttgttc ttgttgctgt tcctgttctg
     3181 aatataattc tggattcatt tttttatcta agtttcctgc taaaccattt agttttggtt
     3241 catcttctgt taacttatta actctttccg caaaggcgcc attcgccatt caggctgcgc
     3301 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     3361 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     3421 aaaacgacgg ccagtgaatt caaaaaaacc ccgccctgtc aggggcgggg tttttgctag
     3481 cttatttata cagttcgtcc ataccgtggg taatgcctgc cgcggtaaca aattccagca
     3541 gtaccatgtg gtcacgtttt tcgttcgggt ctttagacag tttgctctgg taggacaggt
     3601 aatggttatc tggcagcagt actgggccat caccgattgg agtgttctgc tggtagtgat
     3661 ctgccagctg tacgccacca tcttcaatgt tgtgacggat cttgaagttt gctttgatgc
     3721 cgtttttctg cttgtcagca gtaatgtaaa cgttgtggga gttgtagttg tattccagct
     3781 tgtgacccag aatgttgccg tcctctttga agtcaatgcc tttcagttcg atgcggttga
     3841 ccagggtgtc accctcgaat ttaacttcgg cacgggtttt atagttgccg tcgtccttga
     3901 aaaagatcgt acgttcttgg acatagcctt ccggcatggc agatttgaag aagtcatgct
     3961 gtttcatgtg atccgggtaa cgcgcaaaac acatcaggcc gtaacccaga gtagtgacca
     4021 gagttggcca cggaacaggc agtttgccgg tggtacagat cagtttcaga gtcagtttgc
     4081 cataagtagc atcacactcg ccttcaccag acacggaaaa tttgtggcca ttcacgtcac
     4141 catccagctc aaccagaatt ggcacaacgc cggtgaacag ttcttcacct ttgctcataa
     4201 tgtgactttc ctccttaagc ttttaatttc tcatttttca attaaaacaa taaaacatcc
     4261 atcagtgtct tgctctcaca tgttgatgat agtcctttct gctatgatgt ctctgtatgc
     4321 gatttgcatt caaaaatgaa caacaaaaaa cacagaggca aacggatgcc cctgtgccat
     4381 aacatcgaac acaggtttca ttcatctcct ctcacgaacg cttacgaggt tagctgacgg
     4441 attcgggcat atgagagtta gccctaccta aaaaggattc accccaacgg caaaaagccg
     4501 ttctattggg tcccccgctc tgatttcaga ttaagcgatt tgttttctgc acacacatta
     4561 tgccacacct tgtagataaa gtcaacaact ttttcgaaaa tttttcagga attttagcag
     4621 aggttgttct ggatgtagaa caaaacatct ttccgctctt gtgctgtaag gatatctttc
     4681 ttggaagcta ggtaggcaag ggctacggat cctctagagt cgacctgcag gcatgcagcg
     4741 gccgccagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4801 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4861 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4921 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4981 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5041 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5101 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5161 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5221 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5281 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5341 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5401 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5461 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5521 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5581 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5641 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5701 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5761 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5821 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5881 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5941 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6001 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6061 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6121 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6181 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6241 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6301 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6361 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6421 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6481 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6541 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6601 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6661 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6721 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6781 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6841 cgctcgaggc cgcggaaaac tggtttaagc cgactgcgca aaagacataa tcgattcaca
     6901 aaaaataggc acacgaaaaa caagttaagg gatgcagtt
//
