LOCUS       sequence-456024-        6640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control plasmid encoding yfp downstream of an attenuated 
            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 6640)
  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 6640)
  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-yfp
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3126..3148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3140..3157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3141..3157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4290..4878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4370..4389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5049..5840,5841..5909))
                     /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     5672..5691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5910..6014)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6082..6100
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6138..6160)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccatacttaa tatagaaata tcaacttctg ccattaaaag taatgccaat gagcgttttg
       61 tatttaataa tcttttagca aacccgtatt ccacgattaa ataaatctca ttagctatac
      121 tatcaaaaac aattttgcgt attatatccg tacttatgtt ataaggtata ttaccatata
      181 ttttatagga ttggttttta ggaaatttaa actgcaatat atccttgttt aaaacttgga
      241 aattatcgtg atcaacaagt ttattttctg tagttttgca taatttatgg tctatttcaa
      301 tggcagttac gaaattacac ctctttacta attcaagggt aaaatggcct tttcctgagc
      361 cgatttcaaa gatattatca tgttcattta atcttatatt tgtcattatt ttatctatat
      421 tatgttttga agtaataaag ttttgactgt gttttatatt tttctcgttc attataaccc
      481 tctttaattt ggttatatga attttgctta ttaacgattc attataacca cttatttttt
      541 gtttggttga taatgaactg tgctgattac aaaaatacta aaaatgccca tattttttcc
      601 tccttataaa attagtataa ttatagcacg agctctgata aatatgaaca tgatgagtga
      661 tcgttaaatt tatactgcaa tcggatgcga ttattgaata aaagatatga gagatttatc
      721 taatttcttt tttcttgtaa aaaaagaaag ttcttaaagg ttttatagtt ttggtcgtag
      781 agcacacggt ttaacgactt aattacgaag taaataagtc tagtgtgtta gactttatga
      841 aatctatata cgtttatata tatttattat ccggaggtgt agcatgtctc attcaattcc
      901 taggtgggcc caataaaagc aatcaatgaa ccaagacagc tcaaactaat accaataatt
      961 aacaattttt ttatatttat ataatcagat aattttccat atactgctgt tcctatcgaa
     1021 aaagttaaca tatatgcagt gtttacccag tttgtaattc caggagtagt attaaaatga
     1081 tttgcaatat caggtaaaga aacatttaaa accatttcat ttaatacact aaaaaatgaa
     1141 agaatacaaa gccaaaataa aacgctataa aacaaacctt taaatttttt atataaacta
     1201 aacaatttta cctctttact tgaggtgact aaagtttata ggggtaaatt attctttcac
     1261 ctcatataaa ttccccaaat ttttaaatag acacttcata aaaaaatcct cctaaattag
     1321 aatagtttaa atatatcatt tttgttcagt aatattaata tgtacactat ttccaaaatt
     1381 taaattcatg ttgccaaaaa tcgatttgtt tttgcaattg tttttcgttc gcttttaaag
     1441 tcgatttcat taagtccgtt aaatcgactg gcgaaatttc ttttatcaaa ttcttatatt
     1501 ttgttctaga atttctgtga agctttcccc attcttcttc atcactcaat aacataaaaa
     1561 ctattgctct atccgcagtg cgttggatag ttttccaatc tggttgcaag atatgtaaat
     1621 cactaaagca atcattccag taatccacca tatctctttt aagttcgatt tctacacgcc
     1681 ataaatgttc agacataact tcagcatctg cattatcttt acgttcttgc tttttattat
     1741 aaattctaat aaatctatta ctatctctca cgccaaaata ttttgtttct ggcttaccat
     1801 tacgaccata aaaaatagtt ttcttaactg ctttatcaga cattgcatag tagtcactca
     1861 aatcatcttc aaaatcaaag gctaaatcta atcttgtaaa accgtcatct tccatgtagc
     1921 ttattatatt ttgttttaac caaatcattt catctcgtgt aagtttgttt ggattaaatt
     1981 caatacgcat attacgtcta tcccaactat ctgctttcac tttgtcatat tcaatataaa
     2041 ctttttcttg aagtgcttta gctttaaact ttgtttgaag aatatcccaa agtcttattt
     2101 ggggctctac actcataaat ttagaaaggg cttgagcgtt gtctcggttg agatttccaa
     2161 cgatcgtcat tgcatcaaaa cttaattttg gatcagatat gcacacagta tgtgcgtcca
     2221 accggctatt agagtagccg gttttagaaa aattgtctaa atcgtgattt tccaaatgat
     2281 ttgaatgatt tgcatgattg tttttataca taaaaaatcg actccttaat ctcaatttcg
     2341 tttaaggaat cgctcaccca aatatatatc ttgatgtata tttaaatatc gtttaatatc
     2401 taaatataca agattataaa aacaactcag tgtttttttc tttgaatgat gtcgttcaca
     2461 aactttggtc agggcgtgag cgactccttt ttattttgtt attaatataa cactatcaaa
     2521 agatttggtc taatcagatc aagtcttttt ttatttaagc atttgtatta tctggtaaac
     2581 agttaaaact actaaaacac caagtacata cttacttgtt ataaaattct caaaggcttc
     2641 tctgaatttc atttcttcaa ctccttttat tgctttattc caatttcctt attggtcgga
     2701 acctacaggg ctttatgtgt tgttgagttg gtactttctt gggattaatc ccaattcaag
     2761 tccaaccaac tcgctaacaa gttagctaac acatagccca ttccaaccaa taagttttct
     2821 cggcataaat gcatgttcta taagtgcata cctctatctc gtttttgatt cttttgttgt
     2881 tcttgttgct gttcctgttc tgaatataat tctggattca tttttttatc taagtttcct
     2941 gctaaaccat ttagttttgg ttcatcttct gttaacttat taactctttc cgcaaaggcg
     3001 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3061 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcaaaaaaa ccccgccctg
     3181 tcaggggcgg ggtttttgct agcttattta tacagttcgt ccataccgtg ggtaatgcct
     3241 gccgcggtaa caaattccag cagtaccatg tggtcacgtt tttcgttcgg gtctttagac
     3301 agtttgctct ggtaggacag gtaatggtta tctggcagca gtactgggcc atcaccgatt
     3361 ggagtgttct gctggtagtg atctgccagc tgtacgccac catcttcaat gttgtgacgg
     3421 atcttgaagt ttgctttgat gccgtttttc tgcttgtcag cagtaatgta aacgttgtgg
     3481 gagttgtagt tgtattccag cttgtgaccc agaatgttgc cgtcctcttt gaagtcaatg
     3541 cctttcagtt cgatgcggtt gaccagggtg tcaccctcga atttaacttc ggcacgggtt
     3601 ttatagttgc cgtcgtcctt gaaaaagatc gtacgttctt ggacatagcc ttccggcatg
     3661 gcagatttga agaagtcatg ctgtttcatg tgatccgggt aacgcgcaaa acacatcagg
     3721 ccgtaaccca gagtagtgac cagagttggc cacggaacag gcagtttgcc ggtggtacag
     3781 atcagtttca gagtcagttt gccataagta gcatcacact cgccttcacc agacacggaa
     3841 aatttgtggc cattcacgtc accatccagc tcaaccagaa ttggcacaac gccggtgaac
     3901 agttcttcac ctttgctcat aatgtgactt tcctccttaa gcttttggtt cccccgttcg
     3961 tgtttgatta tgcacacaca ttatgccaca ccttgtagat aaagtcaaca actttttcga
     4021 aaatttttca ggaattttag cagaggttgt tctggatgta gaacaaaaca tctttccgct
     4081 cttgtgctgt aaggatatct ttcttggaag ctaggtaggc aagggctacg gatcctctag
     4141 agtcgacctg caggcatgca gcggccgcca gctcactcaa aggcggtaat acggttatcc
     4201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4861 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4981 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5041 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5101 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5161 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5221 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5281 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5341 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5401 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5461 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5521 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5581 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5641 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5701 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5761 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5821 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5881 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5941 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6001 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6061 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6121 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6181 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6241 cggggctggc ttaactatgc ggccgctcga ggccgcggaa aactggttta agccgactgc
     6301 gcaaaagaca taatcgattc acaaaaaata ggcacacgaa aaacaagtta agggatgcag
     6361 tttatgcatc ccttaactta cttattaaat aatttatagc tattgaaaag agataagaat
     6421 tgttcaaagc taatattgtt taaatcgtca attcctgcat gttttaagga attgttaaat
     6481 tgattttttg taaatatttt cttgtattct ttgttaaccc atttcataac gaaataatta
     6541 tacttttgtt tatctttgtg tgatattctt gatttttttc tacttaatct gataagtgag
     6601 ctattcactt taggtttagg atgaaaatat tctcttggaa
//
