LOCUS       sequence-456059-        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     2255..2277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2315..2333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2401..2505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2506..2574,2575..3366)
                     /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(2724..2743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3537..4125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4026..4045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5557..5574)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5557..5573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5566..5588)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ggcgtgagag atagtaatag atttattaga atttataata aaaagcaaga acgtaaagat
       61 aatgcagatg ctgaagttat gtctgaacat ttatggcgtg tagaaatcga acttaaaaga
      121 gatatggtgg attactggaa tgattgcttt agtgatttac atatcttgca accagattgg
      181 aaaactatcc aacgcactgc ggatagagca atagttttta tgttattgag tgatgaagaa
      241 gaatggggaa agcttcacag aaattctaga acaaaatata agaatttgat aaaagaaatt
      301 tcgccagtcg atttaacgga cttaatgaaa tcgactttaa aagcgaacga aaaacaattg
      361 caaaaacaaa tcgatttttg gcaacatgaa tttaaatttt ggaaatagtg tacatattaa
      421 tattactgaa caaaaatgat atatttaaac tattctaatt taggaggatt tttttatgaa
      481 gtgtctattt aaaaatttgg ggaatttata tgaggtgaaa gaataattta cccctataaa
      541 ctttagtcac ctcaagtaaa gaggtaaaat tgtttagttt atataaaaaa tttaaaggtt
      601 tgttttatag cgttttattt tggctttgta ttctttcatt ttttagtgta ttaaatgaaa
      661 tggttttaaa tgtttcttta cctgatattg caaatcattt taatactact cctggaatta
      721 caaactgggt aaacactgca tatatgttaa ctttttcgat aggaacagca gtatatggaa
      781 aattatctga ttatataaat ataaaaaaat tgttaattat tggtattagt ttgagctgtc
      841 ttggttcatt gattgctttt attgggccca cctaggaatt gaatgagaca tgctacacct
      901 ccggataata aatatatata aacgtatata gatttcataa agtctaacac actagactta
      961 tttacttcgt aattaagtcg ttaaaccgtg tgctctacga ccaaaactat aaaaccttta
     1021 agaactttct ttttttacaa gaaaaaagaa attagataaa tctctcatat cttttattca
     1081 ataatcgcat ccgattgcag tataaattta acgatcactc atcatgttca tatttatcag
     1141 agctcgtgct ataattatac taattttata aggaggaaaa aatatgggca tttttagtat
     1201 ttttgtaatc agcacagttc attatcaacc aaacaaaaaa taagtggtta taatgaatcg
     1261 ttaataagca aaattcatat aaccaaatta aagagggtta taatgaacga gaaaaatata
     1321 aaacacagtc aaaactttat tacttcaaaa cataatatag ataaaataat gacaaatata
     1381 agattaaatg aacatgataa tatctttgaa atcggctcag gaaaaggcca ttttaccctt
     1441 gaattagtaa agaggtgtaa tttcgtaact gccattgaaa tagaccataa attatgcaaa
     1501 actacagaaa ataaacttgt tgatcacgat aatttccaag ttttaaacaa ggatatattg
     1561 cagtttaaat ttcctaaaaa ccaatcctat aaaatatatg gtaatatacc ttataacata
     1621 agtacggata taatacgcaa aattgttttt gatagtatag ctaatgagat ttatttaatc
     1681 gtggaatacg ggtttgctaa aagattatta aatacaaaac gctcattggc attactttta
     1741 atggcagaag ttgatatttc tatattaagt atggttccaa gagaatattt tcatcctaaa
     1801 cctaaagtga atagctcact tatcagatta agtagaaaaa aatcaagaat atcacacaaa
     1861 gataaacaaa agtataatta tttcgttatg aaatgggtta acaaagaata caagaaaata
     1921 tttacaaaaa atcaatttaa caattcctta aaacatgcag gaattgacga tttaaacaat
     1981 attagctttg aacaattctt atctcttttc aatagctata aattatttaa taagtaagtt
     2041 aagggatgca taaactgcat cccttaactt gtttttcgtg tgcctatttt ttgtgaatcg
     2101 attatgtctt ttgcgcagtc ggcttaaacc agttttccgc ggcctcgagc ggccgcatag
     2161 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2221 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2281 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     2341 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2401 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2461 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2521 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2581 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2641 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2701 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2761 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2821 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2881 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2941 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3001 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3061 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3121 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3181 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3241 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3301 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3421 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3481 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3541 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3601 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3661 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3721 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3781 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3841 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3901 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3961 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4021 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4081 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4141 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4201 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctggcgg ccgctgcatg
     4261 cctgcaggtc gactctagag gatccgtagc ccttgcctac ctagcttcca agaaagatat
     4321 ccttacagca caagagcgga aagatgtttt gttctacatc cagaacaacc tctgctaaaa
     4381 ttcctgaaaa attttcgaaa aagttgttga ctttatctac aaggtgtggc ataatgtgtg
     4441 tgcagaaaac aaatcgctta atctgaaatc agagcggggg acccaataga acggcttttt
     4501 gccgttgggg tgaatccttt ttaggtaggg ctaactctca tatgcccgaa tccgtcagct
     4561 aacctcgtaa gcgttcgtga gaggagatga atgaaacctg tgttcgatgt tatggcacag
     4621 gggcatccgt ttgcctctgt gttttttgtt gttcattttt gaatgcaaat cgcatacaga
     4681 gacatcatag cagaaaggac tatcatcaac atgtgagagc aagacactga tggatgtttt
     4741 attgttttaa ttgaaaaatg agaaattaaa agcttaagga ggaaagtcac attatgagca
     4801 aaggtgaaga actgttcacc ggcgttgtgc caattctggt tgagctggat ggtgacgtga
     4861 atggccacaa attttccgtg tctggtgaag gcgagtgtga tgctacttat ggcaaactga
     4921 ctctgaaact gatctgtacc accggcaaac tgcctgttcc gtggccaact ctggtcacta
     4981 ctctgggtta cggcctgatg tgttttgcgc gttacccgga tcacatgaaa cagcatgact
     5041 tcttcaaatc tgccatgccg gaaggctatg tccaagaacg tacgatcttt ttcaaggacg
     5101 acggcaacta taaaacccgt gccgaagtta aattcgaggg tgacaccctg gtcaaccgca
     5161 tcgaactgaa aggcattgac ttcaaagagg acggcaacat tctgggtcac aagctggaat
     5221 acaactacaa ctcccacaac gtttacatta ctgctgacaa gcagaaaaac ggcatcaaag
     5281 caaacttcaa gatccgtcac aacattgaag atggtggcgt acagctggca gatcactacc
     5341 agcagaacac tccaatcggt gatggcccag tactgctgcc agataaccat tacctgtcct
     5401 accagagcaa actgtctaaa gacccgaacg aaaaacgtga ccacatggta ctgctggaat
     5461 ttgttaccgc ggcaggcatt acccacggta tggacgaact gtataaataa gctagcaaaa
     5521 accccgcccc tgacagggcg gggttttttt gaattcactg gccgtcgttt tacaacgtcg
     5581 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5641 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     5701 gaatggcgaa tggcgccttt gcggaaagag ttaataagtt aacagaagat gaaccaaaac
     5761 taaatggttt agcaggaaac ttagataaaa aaatgaatcc agaattatat tcagaacagg
     5821 aacagcaaca agaacaacaa aagaatcaaa aacgagatag aggtatgcac ttatagaaca
     5881 tgcatttatg ccgagaaaac ttattggttg gaatgggcta tgtgttagct aacttgttag
     5941 cgagttggtt ggacttgaat tgggattaat cccaagaaag taccaactca acaacacata
     6001 aagccctgta ggttccgacc aataaggaaa ttggaataaa gcaataaaag gagttgaaga
     6061 aatgaaattc agagaagcct ttgagaattt tataacaagt aagtatgtac ttggtgtttt
     6121 agtagtttta actgtttacc agataataca aatgcttaaa taaaaaaaga cttgatctga
     6181 ttagaccaaa tcttttgata gtgttatatt aataacaaaa taaaaaggag tcgctcacgc
     6241 cctgaccaaa gtttgtgaac gacatcattc aaagaaaaaa acactgagtt gtttttataa
     6301 tcttgtatat ttagatatta aacgatattt aaatatacat caagatatat atttgggtga
     6361 gcgattcctt aaacgaaatt gagattaagg agtcgatttt ttatgtataa aaacaatcat
     6421 gcaaatcatt caaatcattt ggaaaatcac gatttagaca atttttctaa aaccggctac
     6481 tctaatagcc ggttggacgc acatactgtg tgcatatctg atccaaaatt aagttttgat
     6541 gcaatgacga tcgttggaaa tctcaaccga gacaacgctc aagccctttc taaatttatg
     6601 agtgtagagc cccaaataag actttgggat attcttcaaa caaagtttaa agctaaagca
     6661 cttcaagaaa aagtttatat tgaatatgac aaagtgaaag cagatagttg ggatagacgt
     6721 aatatgcgta ttgaatttaa tccaaacaaa cttacacgag atgaaatgat ttggttaaaa
     6781 caaaatataa taagctacat ggaagatgac ggttttacaa gattagattt agcctttgat
     6841 tttgaagatg atttgagtga ctactatgca atgtctgata aagcagttaa gaaaactatt
     6901 ttttatggtc gtaatggtaa gccagaaaca aaatatttt
//
