LOCUS       sequence-456015-        6806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  c-di-AMP biosensor plasmid. Encodes yfp downstream of 
            c-di-AMP-binding ktrA 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 6806)
  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 6806)
  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-ktrA-yfp
FEATURES             Location/Qualifiers
     source          1..6806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     143..165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(203..221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        289..393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(394..462,463..1254)
                     /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(612..631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1425..2013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1914..1933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3312..3329)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3312..3328)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3321..3343)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gcgcagtcgg cttaaaccag ttttccgcgg cctcgagcgg ccgcatagtt aagccagccc
       61 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
      121 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
      181 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
      241 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
      301 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      361 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      421 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      481 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      541 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      601 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      661 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      721 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      781 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      841 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      901 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      961 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     1021 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     1081 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     1141 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1201 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1261 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1321 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1381 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1441 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1501 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1561 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1621 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1681 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1741 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1801 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1861 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1921 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1981 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2041 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2101 gtggataacc gtattaccgc ctttgagtga gctggcggcc gctgcatgcc tgcaggtcga
     2161 ctctagagga tccgtagccc ttgcctacct agcttccaag aaagatatcc ttacagcaca
     2221 agagcggaaa gatgttttgt tctacatcca gaacaacctc tgctaaaatt cctgaaaaat
     2281 tttcgaaaaa gttgttgact ttatctacaa ggtgtggcat aatgtgtgtg cataatcaaa
     2341 cacgaacggg ggaaccaacg attggctgtt ttattaacag ccttggggtg aatcttacta
     2401 agtaagaggg ggtactctga atccctaatc cgacagctaa cctcgtaggc gtatacagag
     2461 aggaggtcta ttatcggcca ttctttgcaa tggtcttttt tgtttcaaaa acgttaattc
     2521 actaaagctt aaggaggaaa gtcacattat gagcaaaggt gaagaactgt tcaccggcgt
     2581 tgtgccaatt ctggttgagc tggatggtga cgtgaatggc cacaaatttt ccgtgtctgg
     2641 tgaaggcgag tgtgatgcta cttatggcaa actgactctg aaactgatct gtaccaccgg
     2701 caaactgcct gttccgtggc caactctggt cactactctg ggttacggcc tgatgtgttt
     2761 tgcgcgttac ccggatcaca tgaaacagca tgacttcttc aaatctgcca tgccggaagg
     2821 ctatgtccaa gaacgtacga tctttttcaa ggacgacggc aactataaaa cccgtgccga
     2881 agttaaattc gagggtgaca ccctggtcaa ccgcatcgaa ctgaaaggca ttgacttcaa
     2941 agaggacggc aacattctgg gtcacaagct ggaatacaac tacaactccc acaacgttta
     3001 cattactgct gacaagcaga aaaacggcat caaagcaaac ttcaagatcc gtcacaacat
     3061 tgaagatggt ggcgtacagc tggcagatca ctaccagcag aacactccaa tcggtgatgg
     3121 cccagtactg ctgccagata accattacct gtcctaccag agcaaactgt ctaaagaccc
     3181 gaacgaaaaa cgtgaccaca tggtactgct ggaatttgtt accgcggcag gcattaccca
     3241 cggtatggac gaactgtata aataagctag caaaaacccc gcccctgaca gggcggggtt
     3301 tttttgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3361 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3421 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctttgcgga
     3481 aagagttaat aagttaacag aagatgaacc aaaactaaat ggtttagcag gaaacttaga
     3541 taaaaaaatg aatccagaat tatattcaga acaggaacag caacaagaac aacaaaagaa
     3601 tcaaaaacga gatagaggta tgcacttata gaacatgcat ttatgccgag aaaacttatt
     3661 ggttggaatg ggctatgtgt tagctaactt gttagcgagt tggttggact tgaattggga
     3721 ttaatcccaa gaaagtacca actcaacaac acataaagcc ctgtaggttc cgaccaataa
     3781 ggaaattgga ataaagcaat aaaaggagtt gaagaaatga aattcagaga agcctttgag
     3841 aattttataa caagtaagta tgtacttggt gttttagtag ttttaactgt ttaccagata
     3901 atacaaatgc ttaaataaaa aaagacttga tctgattaga ccaaatcttt tgatagtgtt
     3961 atattaataa caaaataaaa aggagtcgct cacgccctga ccaaagtttg tgaacgacat
     4021 cattcaaaga aaaaaacact gagttgtttt tataatcttg tatatttaga tattaaacga
     4081 tatttaaata tacatcaaga tatatatttg ggtgagcgat tccttaaacg aaattgagat
     4141 taaggagtcg attttttatg tataaaaaca atcatgcaaa tcattcaaat catttggaaa
     4201 atcacgattt agacaatttt tctaaaaccg gctactctaa tagccggttg gacgcacata
     4261 ctgtgtgcat atctgatcca aaattaagtt ttgatgcaat gacgatcgtt ggaaatctca
     4321 accgagacaa cgctcaagcc ctttctaaat ttatgagtgt agagccccaa ataagacttt
     4381 gggatattct tcaaacaaag tttaaagcta aagcacttca agaaaaagtt tatattgaat
     4441 atgacaaagt gaaagcagat agttgggata gacgtaatat gcgtattgaa tttaatccaa
     4501 acaaacttac acgagatgaa atgatttggt taaaacaaaa tataataagc tacatggaag
     4561 atgacggttt tacaagatta gatttagcct ttgattttga agatgatttg agtgactact
     4621 atgcaatgtc tgataaagca gttaagaaaa ctatttttta tggtcgtaat ggtaagccag
     4681 aaacaaaata ttttggcgtg agagatagta atagatttat tagaatttat aataaaaagc
     4741 aagaacgtaa agataatgca gatgctgaag ttatgtctga acatttatgg cgtgtagaaa
     4801 tcgaacttaa aagagatatg gtggattact ggaatgattg ctttagtgat ttacatatct
     4861 tgcaaccaga ttggaaaact atccaacgca ctgcggatag agcaatagtt tttatgttat
     4921 tgagtgatga agaagaatgg ggaaagcttc acagaaattc tagaacaaaa tataagaatt
     4981 tgataaaaga aatttcgcca gtcgatttaa cggacttaat gaaatcgact ttaaaagcga
     5041 acgaaaaaca attgcaaaaa caaatcgatt tttggcaaca tgaatttaaa ttttggaaat
     5101 agtgtacata ttaatattac tgaacaaaaa tgatatattt aaactattct aatttaggag
     5161 gattttttta tgaagtgtct atttaaaaat ttggggaatt tatatgaggt gaaagaataa
     5221 tttaccccta taaactttag tcacctcaag taaagaggta aaattgttta gtttatataa
     5281 aaaatttaaa ggtttgtttt atagcgtttt attttggctt tgtattcttt cattttttag
     5341 tgtattaaat gaaatggttt taaatgtttc tttacctgat attgcaaatc attttaatac
     5401 tactcctgga attacaaact gggtaaacac tgcatatatg ttaacttttt cgataggaac
     5461 agcagtatat ggaaaattat ctgattatat aaatataaaa aaattgttaa ttattggtat
     5521 tagtttgagc tgtcttggtt cattgattgc ttttattggg cccacctagg aattgaatga
     5581 gacatgctac acctccggat aataaatata tataaacgta tatagatttc ataaagtcta
     5641 acacactaga cttatttact tcgtaattaa gtcgttaaac cgtgtgctct acgaccaaaa
     5701 ctataaaacc tttaagaact ttcttttttt acaagaaaaa agaaattaga taaatctctc
     5761 atatctttta ttcaataatc gcatccgatt gcagtataaa tttaacgatc actcatcatg
     5821 ttcatattta tcagagctcg tgctataatt atactaattt tataaggagg aaaaaatatg
     5881 ggcattttta gtatttttgt aatcagcaca gttcattatc aaccaaacaa aaaataagtg
     5941 gttataatga atcgttaata agcaaaattc atataaccaa attaaagagg gttataatga
     6001 acgagaaaaa tataaaacac agtcaaaact ttattacttc aaaacataat atagataaaa
     6061 taatgacaaa tataagatta aatgaacatg ataatatctt tgaaatcggc tcaggaaaag
     6121 gccattttac ccttgaatta gtaaagaggt gtaatttcgt aactgccatt gaaatagacc
     6181 ataaattatg caaaactaca gaaaataaac ttgttgatca cgataatttc caagttttaa
     6241 acaaggatat attgcagttt aaatttccta aaaaccaatc ctataaaata tatggtaata
     6301 taccttataa cataagtacg gatataatac gcaaaattgt ttttgatagt atagctaatg
     6361 agatttattt aatcgtggaa tacgggtttg ctaaaagatt attaaataca aaacgctcat
     6421 tggcattact tttaatggca gaagttgata tttctatatt aagtatggtt ccaagagaat
     6481 attttcatcc taaacctaaa gtgaatagct cacttatcag attaagtaga aaaaaatcaa
     6541 gaatatcaca caaagataaa caaaagtata attatttcgt tatgaaatgg gttaacaaag
     6601 aatacaagaa aatatttaca aaaaatcaat ttaacaattc cttaaaacat gcaggaattg
     6661 acgatttaaa caatattagc tttgaacaat tcttatctct tttcaatagc tataaattat
     6721 ttaataagta agttaaggga tgcataaact gcatccctta acttgttttt cgtgtgccta
     6781 ttttttgtga atcgattatg tctttt
//
