LOCUS       sequence-458444-        6676 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 chloramphenicol resistance to S. aureus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6676)
  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 6676)
  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: pCN38-kimA-yfp
FEATURES             Location/Qualifiers
     source          1..6676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1347..1369
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1407..1425)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1493..1597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1598..1666,1667..2458)
                     /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(1816..1835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2629..3217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3118..3137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4649..4666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4649..4665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4658..4680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtgtattaaa tgaaatggtt ttaaatgttt ctttacctga tattgcaaat cattttaata
       61 ctactcctgg aattacaaac tgggtaaaca ctgcatatat gttaactttt tcgataggaa
      121 cagcagtata tggaaaatta tctgattata taaatataaa aaaattgtta attattggta
      181 ttagtttgag ctgtcttggt tcattgattg cttttattgg gcccacctag gtattatcaa
      241 gataagaaag aaaaggattt ttcgctacgc tcaaatcctt taaaaaaaca caaaagacca
      301 cattttttaa tgtggtcttt tattcttcaa ctaaagcacc cattagttca acaaacgaaa
      361 attggataaa gtgggatatt tttaaaatat atatttatgt tacagtaata ttgactttta
      421 aaaaaggatt gattctaatg aagaaagcag acaagtaagc ctcctaaatt cactttagat
      481 aaaaatttag gaggcatatc aaatgaactt taataaaatt gatttagaca attggaagag
      541 aaaagagata tttaatcatt atttgaacca acaaacgact tttagtataa ccacagaaat
      601 tgatattagt gttttatacc gaaacataaa acaagaagga tataaatttt accctgcatt
      661 tattttctta gtgacaaggg tgataaactc aaatacagct tttagaactg gttacaatag
      721 cgacggagag ttaggttatt gggataagtt agagccactt tatacaattt ttgatggtgt
      781 atctaaaaca ttctctggta tttggactcc tgtaaagaat gacttcaaag agttttatga
      841 tttatacctt tctgatgtag agaaatataa tggttcgggg aaattgtttc ccaaaacacc
      901 tatacctgaa aatgcttttt ctctttctat tattccatgg acttcattta ctgggtttaa
      961 cttaaatatc aataataata gtaattacct tctacccatt attacagcag gaaaattcat
     1021 taataaaggt aattcaatat atttaccgct atctttacag gtacatcatt ctgtttgtga
     1081 tggttatcat gcaggattgt ttatgaactc tattcaggaa ttgtcagata ggcctaatga
     1141 ctggctttta taatatgaga taatgccgac tgtacttttt acagtcggtt ttctaatgtc
     1201 actaacctgc cccgttagtt gaagaaggcc gcggcctcga gcggccgcat agttaagcca
     1261 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     1321 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     1381 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     1441 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     1501 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1561 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1621 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1681 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1741 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1801 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1861 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1921 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1981 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2041 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2101 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     2161 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2221 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2281 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2341 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2401 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2461 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2521 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2761 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3301 ttctgtggat aaccgtatta ccgcctttga gtgagctggc ggccgctgca tgcctgcagg
     3361 tcgactctag aggatccgta gcccttgcct acctagcttc caagaaagat atccttacag
     3421 cacaagagcg gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa
     3481 aaattttcga aaaagttgtt gactttatct acaaggtgtg gcataatgtg tgtgcagaaa
     3541 acaaatcgct taatctgaaa tcagagcggg ggacccaata gaacggcttt ttgccgttgg
     3601 ggtgaatcct ttttaggtag ggctaactct catatgcccg aatccgtcag ctaacctcgt
     3661 aagcgttcgt gagaggagat gaatgaaacc tgtgttcgat gttatggcac aggggcatcc
     3721 gtttgcctct gtgttttttg ttgttcattt ttgaatgcaa atcgcataca gagacatcat
     3781 agcagaaagg actatcatca acatgtgaga gcaagacact gatggatgtt ttattgtttt
     3841 aattgaaaaa tgagaaatta aaagcttaag gaggaaagtc acattatgag caaaggtgaa
     3901 gaactgttca ccggcgttgt gccaattctg gttgagctgg atggtgacgt gaatggccac
     3961 aaattttccg tgtctggtga aggcgagtgt gatgctactt atggcaaact gactctgaaa
     4021 ctgatctgta ccaccggcaa actgcctgtt ccgtggccaa ctctggtcac tactctgggt
     4081 tacggcctga tgtgttttgc gcgttacccg gatcacatga aacagcatga cttcttcaaa
     4141 tctgccatgc cggaaggcta tgtccaagaa cgtacgatct ttttcaagga cgacggcaac
     4201 tataaaaccc gtgccgaagt taaattcgag ggtgacaccc tggtcaaccg catcgaactg
     4261 aaaggcattg acttcaaaga ggacggcaac attctgggtc acaagctgga atacaactac
     4321 aactcccaca acgtttacat tactgctgac aagcagaaaa acggcatcaa agcaaacttc
     4381 aagatccgtc acaacattga agatggtggc gtacagctgg cagatcacta ccagcagaac
     4441 actccaatcg gtgatggccc agtactgctg ccagataacc attacctgtc ctaccagagc
     4501 aaactgtcta aagacccgaa cgaaaaacgt gaccacatgg tactgctgga atttgttacc
     4561 gcggcaggca ttacccacgg tatggacgaa ctgtataaat aagctagcaa aaaccccgcc
     4621 cctgacaggg cggggttttt ttgaattcac tggccgtcgt tttacaacgt cgtgactggg
     4681 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4741 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4801 aatggcgcct ttgcggaaag agttaataag ttaacagaag atgaaccaaa actaaatggt
     4861 ttagcaggaa acttagataa aaaaatgaat ccagaattat attcagaaca ggaacagcaa
     4921 caagaacaac aaaagaatca aaaacgagat agaggtatgc acttatagaa catgcattta
     4981 tgccgagaaa acttattggt tggaatgggc tatgtgttag ctaacttgtt agcgagttgg
     5041 ttggacttga attgggatta atcccaagaa agtaccaact caacaacaca taaagccctg
     5101 taggttccga ccaataagga aattggaata aagcaataaa aggagttgaa gaaatgaaat
     5161 tcagagaagc ctttgagaat tttataacaa gtaagtatgt acttggtgtt ttagtagttt
     5221 taactgttta ccagataata caaatgctta aataaaaaaa gacttgatct gattagacca
     5281 aatcttttga tagtgttata ttaataacaa aataaaaagg agtcgctcac gccctgacca
     5341 aagtttgtga acgacatcat tcaaagaaaa aaacactgag ttgtttttat aatcttgtat
     5401 atttagatat taaacgatat ttaaatatac atcaagatat atatttgggt gagcgattcc
     5461 ttaaacgaaa ttgagattaa ggagtcgatt ttttatgtat aaaaacaatc atgcaaatca
     5521 ttcaaatcat ttggaaaatc acgatttaga caatttttct aaaaccggct actctaatag
     5581 ccggttggac gcacatactg tgtgcatatc tgatccaaaa ttaagttttg atgcaatgac
     5641 gatcgttgga aatctcaacc gagacaacgc tcaagccctt tctaaattta tgagtgtaga
     5701 gccccaaata agactttggg atattcttca aacaaagttt aaagctaaag cacttcaaga
     5761 aaaagtttat attgaatatg acaaagtgaa agcagatagt tgggatagac gtaatatgcg
     5821 tattgaattt aatccaaaca aacttacacg agatgaaatg atttggttaa aacaaaatat
     5881 aataagctac atggaagatg acggttttac aagattagat ttagcctttg attttgaaga
     5941 tgatttgagt gactactatg caatgtctga taaagcagtt aagaaaacta ttttttatgg
     6001 tcgtaatggt aagccagaaa caaaatattt tggcgtgaga gatagtaata gatttattag
     6061 aatttataat aaaaagcaag aacgtaaaga taatgcagat gctgaagtta tgtctgaaca
     6121 tttatggcgt gtagaaatcg aacttaaaag agatatggtg gattactgga atgattgctt
     6181 tagtgattta catatcttgc aaccagattg gaaaactatc caacgcactg cggatagagc
     6241 aatagttttt atgttattga gtgatgaaga agaatgggga aagcttcaca gaaattctag
     6301 aacaaaatat aagaatttga taaaagaaat ttcgccagtc gatttaacgg acttaatgaa
     6361 atcgacttta aaagcgaacg aaaaacaatt gcaaaaacaa atcgattttt ggcaacatga
     6421 atttaaattt tggaaatagt gtacatatta atattactga acaaaaatga tatatttaaa
     6481 ctattctaat ttaggaggat ttttttatga agtgtctatt taaaaatttg gggaatttat
     6541 atgaggtgaa agaataattt acccctataa actttagtca cctcaagtaa agaggtaaaa
     6601 ttgtttagtt tatataaaaa atttaaaggt ttgttttata gcgttttatt ttggctttgt
     6661 attctttcat ttttta
//
