LOCUS       sequence-456050-        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     2527..2549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2587..2605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2673..2777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2778..2846,2847..3638)
                     /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(2996..3015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3809..4397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4298..4317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5696..5713)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5696..5712)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5705..5727)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atgtataaaa acaatcatgc aaatcattca aatcatttgg aaaatcacga tttagacaat
       61 ttttctaaaa ccggctacta atagccggtt ggacgcacat actgtgtgca tatctgatcc
      121 aaaattaagt tttgatgcaa tgacgatcgt tggaaatctc aaccgagaca acgctcaagc
      181 cctttctaaa tttatgagtg tagagcccca aataagactt tgggatattc ttcaaacaaa
      241 gtttaaagct aaagcacttc aagaaaaagt ttatattgaa tatgacaaag tgaaagcaga
      301 tagttgggat agacgtaata tgcgtattga atttaatcca aacaaactta cacgagatga
      361 aatgatttgg ttaaaacaaa atataataag ctacatggaa gatgacggtt ttacaagatt
      421 agatttagcc tttgattttg aagatgattt gagtgactac tatgcaatgt ctgataaagc
      481 agttaagaaa actatttttt atggtcgtaa tggtaagcca gaaacaaaat attttggcgt
      541 gagagatagt aatagattta ttagaattta taataaaaag caagaacgta aagataatgc
      601 agatgctgaa gttatgtctg aacatttatg gcgtgtagaa atcgaactta aaagagatat
      661 ggtggattac tggaatgatt gctttagtga tttacatatc ttgcaaccag attggaaaac
      721 tatccaacgc actgcggata gagcaatagt ttttatgtta ttgagtgatg aagaagaatg
      781 gggaaagctt cacagaaatt ctagaacaaa atataagaat ttgataaaag aaatttcgcc
      841 agtcgattta acggacttaa tgaaatcgac tttaaaagcg aacgaaaaac aattgcaaaa
      901 acaaatcgat ttttggcaac atgaatttaa attttggaaa tagtgtacat attaatatta
      961 ctgaacaaaa atgatatatt taaactattc taatttagga ggattttttt atgaagtgtc
     1021 tatttaaaaa tttggggaat ttatatgagg tgaaagaata atttacccct ataaacttta
     1081 gtcacctcaa gtaaagaggt aaaattgttt agtttatata aaaaatttaa aggtttgttt
     1141 tatagcgttt tattttggct ttgtattctt tcatttttta gtgtattaaa tgaaatggtt
     1201 ttaaatgttt ctttacctga tattgcaaat cattttaata ctactcctgg aattacaaac
     1261 tgggtaaaca ctgcatatat gttaactttt tcgataggaa cagcagtata tggaaaatta
     1321 tctgattata taaatataaa aaaattgtta attattggta ttagtttgag ctgtcttggt
     1381 tcattgattg cttttattgg gcccacctag gtattatcaa gataagaaag aaaaggattt
     1441 ttcgctacgc tcaaatcctt taaaaaaaca caaaagacca cattttttaa tgtggtcttt
     1501 tattcttcaa ctaaagcacc cattagttca acaaacgaaa attggataaa gtgggatatt
     1561 tttaaaatat atatttatgt tacagtaata ttgactttta aaaaaggatt gattctaatg
     1621 aagaaagcag acaagtaagc ctcctaaatt cactttagat aaaaatttag gaggcatatc
     1681 aaatgaactt taataaaatt gatttagaca attggaagag aaaagagata tttaatcatt
     1741 atttgaacca acaaacgact tttagtataa ccacagaaat tgatattagt gttttatacc
     1801 gaaacataaa acaagaagga tataaatttt accctgcatt tattttctta gtgacaaggg
     1861 tgataaactc aaatacagct tttagaactg gttacaatag cgacggagag ttaggttatt
     1921 gggataagtt agagccactt tatacaattt ttgatggtgt atctaaaaca ttctctggta
     1981 tttggactcc tgtaaagaat gacttcaaag agttttatga tttatacctt tctgatgtag
     2041 agaaatataa tggttcgggg aaattgtttc ccaaaacacc tatacctgaa aatgcttttt
     2101 ctctttctat tattccatgg acttcattta ctgggtttaa cttaaatatc aataataata
     2161 gtaattacct tctacccatt attacagcag gaaaattcat taataaaggt aattcaatat
     2221 atttaccgct atctttacag gtacatcatt ctgtttgtga tggttatcat gcaggattgt
     2281 ttatgaactc tattcaggaa ttgtcagata ggcctaatga ctggctttta taatatgaga
     2341 taatgccgac tgtacttttt acagtcggtt ttctaatgtc actaacctgc cccgttagtt
     2401 gaagaaggcc gcggcctcga gcggccgcat agttaagcca gccccgacac ccgccaacac
     2461 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2521 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2581 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2641 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2701 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2761 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2821 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2881 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2941 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3001 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3061 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3121 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3181 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3241 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3301 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3361 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3421 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3481 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3541 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3601 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3661 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3721 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4381 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4441 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4501 ccgcctttga gtgagctggc ggccgctgca tgcctgcagg tcgactctag aggatccgta
     4561 gcccttgcct acctagcttc caagaaagat atccttacag cacaagagcg gaaagatgtt
     4621 ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttcga aaaagttgtt
     4681 gactttatct acaaggtgtg gcataatgtg tgtgcataat caaacacgaa cgggggaacc
     4741 aacgattggc tgttttatta acagccttgg ggtgaatctt actaagtaag agggggtact
     4801 ctgaatccct aatccgacag ctaacctcgt aggcgtatac agagaggagg tctattatcg
     4861 gccattcttt gcaatggtct tttttgtttc aaaaacgtta attcactaaa gcttaaggag
     4921 gaaagtcaca ttatgagcaa aggtgaagaa ctgttcaccg gcgttgtgcc aattctggtt
     4981 gagctggatg gtgacgtgaa tggccacaaa ttttccgtgt ctggtgaagg cgagtgtgat
     5041 gctacttatg gcaaactgac tctgaaactg atctgtacca ccggcaaact gcctgttccg
     5101 tggccaactc tggtcactac tctgggttac ggcctgatgt gttttgcgcg ttacccggat
     5161 cacatgaaac agcatgactt cttcaaatct gccatgccgg aaggctatgt ccaagaacgt
     5221 acgatctttt tcaaggacga cggcaactat aaaacccgtg ccgaagttaa attcgagggt
     5281 gacaccctgg tcaaccgcat cgaactgaaa ggcattgact tcaaagagga cggcaacatt
     5341 ctgggtcaca agctggaata caactacaac tcccacaacg tttacattac tgctgacaag
     5401 cagaaaaacg gcatcaaagc aaacttcaag atccgtcaca acattgaaga tggtggcgta
     5461 cagctggcag atcactacca gcagaacact ccaatcggtg atggcccagt actgctgcca
     5521 gataaccatt acctgtccta ccagagcaaa ctgtctaaag acccgaacga aaaacgtgac
     5581 cacatggtac tgctggaatt tgttaccgcg gcaggcatta cccacggtat ggacgaactg
     5641 tataaataag ctagcaaaaa ccccgcccct gacagggcgg ggtttttttg aattcactgg
     5701 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5761 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5821 cccaacagtt gcgcagcctg aatggcgaat ggcgcctttg cggaaagagt taataagtta
     5881 acagaagatg aaccaaaact aaatggttta gcaggaaact tagataaaaa aatgaatcca
     5941 gaattatatt cagaacagga acagcaacaa gaacaacaaa agaatcaaaa acgagataga
     6001 ggtatgcact tatagaacat gcatttatgc cgagaaaact tattggttgg aatgggctat
     6061 gtgttagcta acttgttagc gagttggttg gacttgaatt gggattaatc ccaagaaagt
     6121 accaactcaa caacacataa agccctgtag gttccgacca ataaggaaat tggaataaag
     6181 caataaaagg agttgaagaa atgaaattca gagaagcctt tgagaatttt ataacaagta
     6241 agtatgtact tggtgtttta gtagttttaa ctgtttacca gataatacaa atgcttaaat
     6301 aaaaaaagac ttgatctgat tagaccaaat cttttgatag tgttatatta ataacaaaat
     6361 aaaaaggagt cgctcacgcc ctgaccaaag tttgtgaacg acatcattca aagaaaaaaa
     6421 cactgagttg tttttataat cttgtatatt tagatattaa acgatattta aatatacatc
     6481 aagatatata tttgggtgag cgattcctta aacgaaattg agattaagga gtcgattttt
     6541 t
//
