LOCUS       sequence-456061-        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     2792..2814
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2852..2870)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2938..3042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3043..3111,3112..3903)
                     /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(3261..3280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4074..4662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4563..4582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5961..5978)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5961..5977)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5970..5992)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atgtataaaa acaatcatgc aaatcattca aatcatttgg aaaatcacga tttagacaat
       61 ttttctaaaa ccggctactc taatagccgg ttggacgcac atactgtgtg catatctgat
      121 ccaaaattaa gttttgatgc aatgacgatc gttggaaatc tcaaccgaga caacgctcaa
      181 gccctttcta aatttatgag tgtagagccc caaataagac tttgggatat tcttcaaaca
      241 aagtttaaag ctaaagcact tcaagaaaaa gtttatattg aatatgacaa agtgaaagca
      301 gatagttggg atagacgtaa tatgcgtatt gaatttaatc caaacaaact tacacgagat
      361 gaaatgattt ggttaaaaca aaatataata agctacatgg aagatgacgg ttttacaaga
      421 ttagatttag cctttgattt tgaagatgat ttgagtgact actatgcaat gtctgataaa
      481 gcagttaaga aaactatttt ttatggtcgt aatggtaagc cagaaacaaa atattttggc
      541 gtgagagata gtaatagatt tattagaatt tataataaaa agcaagaacg taaagataat
      601 gcagatgctg aagttatgtc tgaacattta tggcgtgtag aaatcgaact taaaagagat
      661 atggtggatt actggaatga ttgctttagt gatttacata tcttgcaacc agattggaaa
      721 actatccaac gcactgcgga tagagcaata gtttttatgt tattgagtga tgaagaagaa
      781 tggggaaagc ttcacagaaa ttctagaaca aaatataaga atttgataaa agaaatttcg
      841 ccagtcgatt taacggactt aatgaaatcg actttaaaag cgaacgaaaa acaattgcaa
      901 aaacaaatcg atttttggca acatgaattt aaattttgga aatagtgtac atattaatat
      961 tactgaacaa aaatgatata tttaaactat tctaatttag gaggattttt ttatgaagtg
     1021 tctatttaaa aatttgggga atttatatga ggtgaaagaa taatttaccc ctataaactt
     1081 tagtcacctc aagtaaagag gtaaaattgt ttagtttata taaaaaattt aaaggtttgt
     1141 tttatagcgt tttattttgg ctttgtattc tttcattttt tagtgtatta aatgaaatgg
     1201 ttttaaatgt ttctttacct gatattgcaa atcattttaa tactactcct ggaattacaa
     1261 actgggtaaa cactgcatat atgttaactt tttcgatagg aacagcagta tatggaaaat
     1321 tatctgatta tataaatata aaaaaattgt taattattgg tattagtttg agctgtcttg
     1381 gttcattgat tgcttttatt gggcccacct aggaattgaa tgagacatgc tacacctccg
     1441 gataataaat atatataaac gtatatagat ttcataaagt ctaacacact agacttattt
     1501 acttcgtaat taagtcgtta aaccgtgtgc tctacgacca aaactataaa acctttaaga
     1561 actttctttt tttacaagaa aaaagaaatt agataaatct ctcatatctt ttattcaata
     1621 atcgcatccg attgcagtat aaatttaacg atcactcatc atgttcatat ttatcagagc
     1681 tcgtgctata attatactaa ttttataagg aggaaaaaat atgggcattt ttagtatttt
     1741 tgtaatcagc acagttcatt atcaaccaaa caaaaaataa gtggttataa tgaatcgtta
     1801 ataagcaaaa ttcatataac caaattaaag agggttataa tgaacgagaa aaatataaaa
     1861 cacagtcaaa actttattac ttcaaaacat aatatagata aaataatgac aaatataaga
     1921 ttaaatgaac atgataatat ctttgaaatc ggctcaggaa aaggccattt tacccttgaa
     1981 ttagtaaaga ggtgtaattt cgtaactgcc attgaaatag accataaatt atgcaaaact
     2041 acagaaaata aacttgttga tcacgataat ttccaagttt taaacaagga tatattgcag
     2101 tttaaatttc ctaaaaacca atcctataaa atatatggta atatacctta taacataagt
     2161 acggatataa tacgcaaaat tgtttttgat agtatagcta atgagattta tttaatcgtg
     2221 gaatacgggt ttgctaaaag attattaaat acaaaacgct cattggcatt acttttaatg
     2281 gcagaagttg atatttctat attaagtatg gttccaagag aatattttca tcctaaacct
     2341 aaagtgaata gctcacttat cagattaagt agaaaaaaat caagaatatc acacaaagat
     2401 aaacaaaagt ataattattt cgttatgaaa tgggttaaca aagaatacaa gaaaatattt
     2461 acaaaaaatc aatttaacaa ttccttaaaa catgcaggaa ttgacgattt aaacaatatt
     2521 agctttgaac aattcttatc tcttttcaat agctataaat tatttaataa gtaagttaag
     2581 ggatgcataa actgcatccc ttaacttgtt tttcgtgtgc ctattttttg tgaatcgatt
     2641 atgtcttttg cgcagtcggc ttaaaccagt tttccgcggc ctcgagcggc cgcatagtta
     2701 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2761 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2821 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2881 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2941 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3001 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3061 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3121 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3181 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3241 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3301 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3361 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3421 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3481 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3541 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3601 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3661 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3721 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3781 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3841 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3901 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3961 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4021 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4081 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4141 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4201 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4261 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4321 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4381 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4441 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4501 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4561 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4621 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4681 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4741 cctgattctg tggataaccg tattaccgcc tttgagtgag ctggcggccg ctgcatgcct
     4801 gcaggtcgac tctagaggat ccgtagccct tgcctaccta gcttccaaga aagatatcct
     4861 tacagcacaa gagcggaaag atgttttgtt ctacatccag aacaacctct gctaaaattc
     4921 ctgaaaaatt ttcgaaaaag ttgttgactt tatctacaag gtgtggcata atgtgtgtgc
     4981 ataatcaaac acgaacgggg gaaccaacga ttggctgttt tattaacagc cttggggtga
     5041 atcttactaa gtaagagggg gtactctgaa tccctaatcc gacagctaac ctcgtaggcg
     5101 tatacagaga ggaggtctat tatcggccat tctttgcaat ggtctttttt gtttcaaaaa
     5161 cgttaattca ctaaagctta aggaggaaag tcacattatg agcaaaggtg aagaactgtt
     5221 caccggcgtt gtgccaattc tggttgagct ggatggtgac gtgaatggcc acaaattttc
     5281 cgtgtctggt gaaggcgagt gtgatgctac ttatggcaaa ctgactctga aactgatctg
     5341 taccaccggc aaactgcctg ttccgtggcc aactctggtc actactctgg gttacggcct
     5401 gatgtgtttt gcgcgttacc cggatcacat gaaacagcat gacttcttca aatctgccat
     5461 gccggaaggc tatgtccaag aacgtacgat ctttttcaag gacgacggca actataaaac
     5521 ccgtgccgaa gttaaattcg agggtgacac cctggtcaac cgcatcgaac tgaaaggcat
     5581 tgacttcaaa gaggacggca acattctggg tcacaagctg gaatacaact acaactccca
     5641 caacgtttac attactgctg acaagcagaa aaacggcatc aaagcaaact tcaagatccg
     5701 tcacaacatt gaagatggtg gcgtacagct ggcagatcac taccagcaga acactccaat
     5761 cggtgatggc ccagtactgc tgccagataa ccattacctg tcctaccaga gcaaactgtc
     5821 taaagacccg aacgaaaaac gtgaccacat ggtactgctg gaatttgtta ccgcggcagg
     5881 cattacccac ggtatggacg aactgtataa ataagctagc aaaaaccccg cccctgacag
     5941 ggcggggttt ttttgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6001 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6061 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     6121 ctttgcggaa agagttaata agttaacaga agatgaacca aaactaaatg gtttagcagg
     6181 aaacttagat aaaaaaatga atccagaatt atattcagaa caggaacagc aacaagaaca
     6241 acaaaagaat caaaaacgag atagaggtat gcacttatag aacatgcatt tatgccgaga
     6301 aaacttattg gttggaatgg gctatgtgtt agctaacttg ttagcgagtt ggttggactt
     6361 gaattgggat taatcccaag aaagtaccaa ctcaacaaca cataaagccc tgtaggttcc
     6421 gaccaataag gaaattggaa taaagcaata aaaggagttg aagaaatgaa attcagagaa
     6481 gcctttgaga attttataac aagtaagtat gtacttggtg ttttagtagt tttaactgtt
     6541 taccagataa tacaaatgct taaataaaaa aagacttgat ctgattagac caaatctttt
     6601 gatagtgtta tattaataac aaaataaaaa ggagtcgctc acgccctgac caaagtttgt
     6661 gaacgacatc attcaaagaa aaaaacactg agttgttttt ataatcttgt atatttagat
     6721 attaaacgat atttaaatat acatcaagat atatatttgg gtgagcgatt ccttaaacga
     6781 aattgagatt aaggagtcga tttttt
//
