LOCUS       sequence-456013-        6541 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 chloramphenicol resistance to S. aureus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6541)
  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 6541)
  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-ktrA-yfp
FEATURES             Location/Qualifiers
     source          1..6541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3077..3099
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3091..3108
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3092..3108
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4407..4995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4487..4506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5166..5957,5958..6026))
                     /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     5789..5808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6027..6131)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6199..6217
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6255..6277)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 catcacaaac agaatgatgt acctgtaaag atagcggtaa atatattgaa ttacctttat
       61 taatgaattt tcctgctgta ataatgggta gaaggtaatt actattatta ttgatattta
      121 agttaaaccc agtaaatgaa gtccatggaa taatagaaag agaaaaagca ttttcaggta
      181 taggtgtttt gggaaacaat ttccccgaac cattatattt ctctacatca gaaaggtata
      241 aatcataaaa ctctttgaag tcattcttta caggagtcca aataccagag aatgttttag
      301 atacaccatc aaaaattgta taaagtggct ctaacttatc ccaataacct aactctccgt
      361 cgctattgta accagttcta aaagctgtat ttgagtttat cacccttgtc actaagaaaa
      421 taaatgcagg gtaaaattta tatccttctt gttttatgtt tcggtataaa acactaatat
      481 caatttctgt ggttatacta aaagtcgttt gttggttcaa ataatgatta aatatctctt
      541 ttctcttcca attgtctaaa tcaattttat taaagttcat ttgatatgcc tcctaaattt
      601 ttatctaaag tgaatttagg aggcttactt gtctgctttc ttcattagaa tcaatccttt
      661 tttaaaagtc aatattactg taacataaat atatatttta aaaatatccc actttatcca
      721 attttcgttt gttgaactaa tgggtgcttt agttgaagaa taaaagacca cattaaaaaa
      781 tgtggtcttt tgtgtttttt taaaggattt gagcgtagcg aaaaatcctt ttctttctta
      841 tcttgataat acctaggtgg gcccaataaa agcaatcaat gaaccaagac agctcaaact
      901 aataccaata attaacaatt tttttatatt tatataatca gataattttc catatactgc
      961 tgttcctatc gaaaaagtta acatatatgc agtgtttacc cagtttgtaa ttccaggagt
     1021 agtattaaaa tgatttgcaa tatcaggtaa agaaacattt aaaaccattt catttaatac
     1081 actaaaaaat gaaagaatac aaagccaaaa taaaacgcta taaaacaaac ctttaaattt
     1141 tttatataaa ctaaacaatt ttacctcttt acttgaggtg actaaagttt ataggggtaa
     1201 attattcttt cacctcatat aaattcccca aatttttaaa tagacacttc ataaaaaaat
     1261 cctcctaaat tagaatagtt taaatatatc atttttgttc agtaatatta atatgtacac
     1321 tatttccaaa atttaaattc atgttgccaa aaatcgattt gtttttgcaa ttgtttttcg
     1381 ttcgctttta aagtcgattt cattaagtcc gttaaatcga ctggcgaaat ttcttttatc
     1441 aaattcttat attttgttct agaatttctg tgaagctttc cccattcttc ttcatcactc
     1501 aataacataa aaactattgc tctatccgca gtgcgttgga tagttttcca atctggttgc
     1561 aagatatgta aatcactaaa gcaatcattc cagtaatcca ccatatctct tttaagttcg
     1621 atttctacac gccataaatg ttcagacata acttcagcat ctgcattatc tttacgttct
     1681 tgctttttat tataaattct aataaatcta ttactatctc tcacgccaaa atattttgtt
     1741 tctggcttac cattacgacc ataaaaaata gttttcttaa ctgctttatc agacattgca
     1801 tagtagtcac tcaaatcatc ttcaaaatca aaggctaaat ctaatcttgt aaaaccgtca
     1861 tcttccatgt agcttattat attttgtttt aaccaaatca tttcatctcg tgtaagtttg
     1921 tttggattaa attcaatacg catattacgt ctatcccaac tatctgcttt cactttgtca
     1981 tattcaatat aaactttttc ttgaagtgct ttagctttaa actttgtttg aagaatatcc
     2041 caaagtctta tttggggctc tacactcata aatttagaaa gggcttgagc gttgtctcgg
     2101 ttgagatttc caacgatcgt cattgcatca aaacttaatt ttggatcaga tatgcacaca
     2161 gtatgtgcgt ccaaccggct attagtagcc ggttttagaa aaattgtcta aatcgtgatt
     2221 ttccaaatga tttgaatgat ttgcatgatt gtttttatac ataaaaaatc gactccttaa
     2281 tctcaatttc gtttaaggaa tcgctcaccc aaatatatat cttgatgtat atttaaatat
     2341 cgtttaatat ctaaatatac aagattataa aaacaactca gtgttttttt ctttgaatga
     2401 tgtcgttcac aaactttggt cagggcgtga gcgactcctt tttattttgt tattaatata
     2461 acactatcaa aagatttggt ctaatcagat caagtctttt tttatttaag catttgtatt
     2521 atctggtaaa cagttaaaac tactaaaaca ccaagtacat acttacttgt tataaaattc
     2581 tcaaaggctt ctctgaattt catttcttca actcctttta ttgctttatt ccaatttcct
     2641 tattggtcgg aacctacagg gctttatgtg ttgttgagtt ggtactttct tgggattaat
     2701 cccaattcaa gtccaaccaa ctcgctaaca agttagctaa cacatagccc attccaacca
     2761 ataagttttc tcggcataaa tgcatgttct ataagtgcat acctctatct cgtttttgat
     2821 tcttttgttg ttcttgttgc tgttcctgtt ctgaatataa ttctggattc atttttttat
     2881 ctaagtttcc tgctaaacca tttagttttg gttcatcttc tgttaactta ttaactcttt
     2941 ccgcaaaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     3001 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     3061 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcaaaaaa
     3121 accccgccct gtcaggggcg gggtttttgc tagcttattt atacagttcg tccataccgt
     3181 gggtaatgcc tgccgcggta acaaattcca gcagtaccat gtggtcacgt ttttcgttcg
     3241 ggtctttaga cagtttgctc tggtaggaca ggtaatggtt atctggcagc agtactgggc
     3301 catcaccgat tggagtgttc tgctggtagt gatctgccag ctgtacgcca ccatcttcaa
     3361 tgttgtgacg gatcttgaag tttgctttga tgccgttttt ctgcttgtca gcagtaatgt
     3421 aaacgttgtg ggagttgtag ttgtattcca gcttgtgacc cagaatgttg ccgtcctctt
     3481 tgaagtcaat gcctttcagt tcgatgcggt tgaccagggt gtcaccctcg aatttaactt
     3541 cggcacgggt tttatagttg ccgtcgtcct tgaaaaagat cgtacgttct tggacatagc
     3601 cttccggcat ggcagatttg aagaagtcat gctgtttcat gtgatccggg taacgcgcaa
     3661 aacacatcag gccgtaaccc agagtagtga ccagagttgg ccacggaaca ggcagtttgc
     3721 cggtggtaca gatcagtttc agagtcagtt tgccataagt agcatcacac tcgccttcac
     3781 cagacacgga aaatttgtgg ccattcacgt caccatccag ctcaaccaga attggcacaa
     3841 cgccggtgaa cagttcttca cctttgctca taatgtgact ttcctcctta agctttagtg
     3901 aattaacgtt tttgaaacaa aaaagaccat tgcaaagaat ggccgataat agacctcctc
     3961 tctgtatacg cctacgaggt tagctgtcgg attagggatt cagagtaccc cctcttactt
     4021 agtaagattc accccaaggc tgttaataaa acagccaatc gttggttccc ccgttcgtgt
     4081 ttgattatgc acacacatta tgccacacct tgtagataaa gtcaacaact ttttcgaaaa
     4141 tttttcagga attttagcag aggttgttct ggatgtagaa caaaacatct ttccgctctt
     4201 gtgctgtaag gatatctttc ttggaagcta ggtaggcaag ggctacggat cctctagagt
     4261 cgacctgcag gcatgcagcg gccgccagct cactcaaagg cggtaatacg gttatccaca
     4321 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4381 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4441 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4501 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4561 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4621 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4681 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4741 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4801 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4861 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4921 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4981 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5041 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5101 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5161 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5221 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5281 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5341 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5401 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5461 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5521 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5581 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5641 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5701 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5761 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5821 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5881 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5941 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6001 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6061 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6121 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6181 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     6241 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     6301 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     6361 ggctggctta actatgcggc cgctcgaggc cgcggccttc ttcaactaac ggggcaggtt
     6421 agtgacatta gaaaaccgac tgtaaaaagt acagtcggca ttatctcata ttataaaagc
     6481 cagtcattag gcctatctga caattcctga atagagttca taaacaatcc tgcatgataa
     6541 c
//
