LOCUS       sequence-112123-        6069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium Repressor plasmid for two-strain system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6069)
  AUTHORS   Chen Y, Kim JK, Hirning AJ, Josic K, Bennett MR
  TITLE     SYNTHETIC BIOLOGY. Emergent genetic oscillations in a synthetic 
            microbial consortium.
  JOURNAL   Science. 2015 Aug 28;349(6251):986-9. doi: 10.1126/science.aaa3794.
  PUBMED    26315440
REFERENCE   2  (bases 1 to 6069)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2015-08-28- 28"; volume: "349"; 
            issue: "6251"; pages: "986-9"
COMMENT     Sequence Label: pC236
FEATURES             Location/Qualifiers
     source          1..6069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      158..229
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             439..630
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(624..646)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    732..874
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     787..806
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(889..906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1060..1648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1140..1159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1759..1778)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1770..2585
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1770..2585
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1843..1862)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2734..2761)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    3694..3710
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             complement(3895..3924)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    4126..4142
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    4784..4800
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             5002..5031
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    5216..5232
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             5317..6030
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 gagctcgaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
       61 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
      121 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
      181 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
      241 taagtgcgga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
      301 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
      361 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
      421 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
      481 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
      541 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
      601 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
      661 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      721 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
      781 actatgcggc atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc
      841 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
      901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1501 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg
     1741 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     1801 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1861 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     1921 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1981 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2041 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
     2101 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2161 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2221 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2281 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2341 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2401 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2461 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2521 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2581 tctaagaatt aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2641 ggggttccgc gcacatttcc ccggctagct aggaaataat aaaaaagccg gattaataat
     2701 ctggcttttt atattctctc tctagtatat aaacgcagaa aggcccaccc gaaggtgagc
     2761 cagtgtgaga cagcggtgcg gactgttgta actcagaata agaaatgagg ccgctcatgg
     2821 cgttgactct cagtcatagt atcgtggtat caccggttgg ttccactctc tgttgcgggc
     2881 aacttcagca gcacgtaggg gaagctttta ttacgctgca agggcgtaat tttcgtcgtt
     2941 cgctgctgct gccatctcca ggaattgggt cacgaccgag cgggccggca cgtgcctgac
     3001 ataaaggggt agggtgaggc cgagggcggc gcgcatcttg cgcagcacgc ggtccgagac
     3061 ttcgaaggcg ccgcagcaga cgggatattt gccgtagccg tctgcgcggc cgagttcttc
     3121 cacctcggcg ccggcgcgct tgtagacgcg cttgaggtag ggctcgtagt tggagatcat
     3181 cgtgtggatg ccgtgatcga gcgcgcattc gcaaagcgcg agcagcatca tggagaaggc
     3241 gcggccggcg tcgatctcgg ggaaatcctt ggcgatcgcc tcctcgtcga tgcacatgcg
     3301 cgtgccttcc cagatgccgg gggcgataag atcggcggca tcagggaacg tctcgcggaa
     3361 gacgtcgtag agaagggtcg ggccggtcgt cggcatcagg cgcatgccgc cataaagacg
     3421 ggtgcggctg tcgttgcacc agacgagata ggcgggagca agcgaatcgt agctgtcacg
     3481 ttcgtaaggg ccgatgacag gaacgtccca gcagagcgta tcggcgaaga ccttcttgcg
     3541 cagacgaaac atctggtcga gtacggcagc gtatttctga tactcatgtg cctgaatgat
     3601 aacgaacata ctagtatttc tccttagcat gattaagatg tttcagtacg aaaattgctt
     3661 tcattgttga tctccttttt aagtggcatg caattgttat ccgctcacaa ttgtgctcat
     3721 tatcaattgt gagcgctcac aagtaaaact gccagatttc acaggatcga cagttcatag
     3781 gtgattgctc aggacatttc tgttagaagg aatcgttttc cttacttttc cttacgcaca
     3841 agagttccgt agctgttcaa gtttgtgttt caactgttct cgtcgtttcc gcaacaagtc
     3901 ctcttcagaa atgagctttt gctcctctgc ttggacggac aggatgtatg ctgtggcttt
     3961 tttaaggata actaccttgg gggccttttc attgttttcc aactccggga tctggtcacg
     4021 cagggcaaaa aagctccgtt ttagctcgtt cctcctctgg cgctccaaga cgttgtgtgt
     4081 tcgcctcttg acattctcct cggtgtccga gggccctgtg tgaaattgtt atccgctcac
     4141 aattccacac agacgtcgga attgccagct ggggcgccct ctggtaaggt tgggaagctc
     4201 gagaagcgcg tctgaatagg cttcttgtat tcctgccaca cagggaaacc atcaggagtt
     4261 acccaatgca cagcgcaacg cttgcgaaga atctctccag tcttcttatc tttgacctca
     4321 gcagccagca gcttagcagc agacttaagc cagttcattg cttcaaccgc agctaccacc
     4381 gtcacgctca cagattccca aatcagctta gccatgtatc cagcagcctg attcggctga
     4441 gtgaacatca gacccttgcc ggaatcaata gctggctgaa tggtatcttc cagcacttgt
     4501 tgacggaagc cgaactcttt ggacccgtaa gccagcgtca tgactgaacg cttagtcaca
     4561 ctgcgagtaa caccgtaagc cagccattga ccagccagtg ccttagtgcc cagcttgact
     4621 ttctcagaga tttcaccagt gttctcatcg gtcacggtaa ctacttcgtt atcggtccca
     4681 ttgattgcgt ctgcttgtag aatctcgttg actttcttag caacaatccc gtagatgtcc
     4741 tgaacggttt cactaggaag caagttaaga cgtctgtgtg gaattgtgag cggataacaa
     4801 tttcacacag ggccctcgga caccgaggag aatgtcaaga ggcgaacaca caacgtcttg
     4861 gagcgccaga ggaggaacga gctaaaacgg agcttttttg ccctgcgtga ccagatcccg
     4921 gagttggaaa acaatgaaaa ggcccccaag gtagttatcc ttaaaaaagc cacagcatac
     4981 atcctgtccg tccaagcaga ggagcaaaag ctcatttctg aagaggactt gttgcggaaa
     5041 cgacgagaac agttgaaaca caaacttgaa cagctacgga actcttgtgc gtggcgcatg
     5101 tcgttatcgc aaaaccgctg cacacttttg cgcgacatgc tctgatcccc ctcatctggg
     5161 ggggcctatc tgagggaatt tacttgtgag cgctcacaat tgataatgag cacaattgtg
     5221 agcggataac aattgcatgc cacttaaaaa ggagatcaac aatgaaagca attttcgtac
     5281 tgaaacatct taatcatgct aaggagaaat actagtatgc gtaaaggcga agagctgttc
     5341 actggtgtcg tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc
     5401 gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt
     5461 actactggta aactgccggt accttggccg actctggtaa cgacgctgac ttatggtgtt
     5521 cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg
     5581 ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg
     5641 cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt
     5701 gactttaaag aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac
     5761 aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc
     5821 cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc
     5881 ggtgatggtc ctgttctgct gccagacaat cactatctga gctaccaaag cgttctgtct
     5941 aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc
     6001 atcacgcatg gtatggatga actgtacaaa gcagcgaacg acgaaaatta cgcccttgca
     6061 gcgtaataa
//
