LOCUS       sequence-112068-        6529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium P2 activator plasmid for two-strain system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6529)
  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 6529)
  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: pC220
FEATURES             Location/Qualifiers
     source          1..6529
                     /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(3248..3275)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    4154..4170
                     /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(4355..4384)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    4586..4602
                     /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    5244..5260
                     /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             5462..5491
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    5676..5692
                     /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             5777..6490
                     /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
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     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 ccggctagcc gcctggtgct acgcctgaat aagtgataat
     2701 aagcggatga atggcagaaa ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg
     2761 gtcgttaaat agccgcttat gtctattgct ggtttaccgg tttattgact accggaagca
     2821 gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc tcaggctctc cccgtggagg
     2881 taataattga cgatatgatc atttattctg cctcccagag cctgataaaa acggttagcg
     2941 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     3001 ccggaacata atggtgcagg gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
     3061 cgggggactg ttgggcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     3121 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctac cgcctgcggc
     3181 cgctaggaaa taataaaaaa gccggattaa taatctggct ttttatattc tctctctagt
     3241 atataaacgc agaaaggccc acccgaaggt gagccagtgt gagacagcgg tgcggactgt
     3301 tgtaactcag aataagaaat gaggccgctc atggcgttga ctctcagtca tagtatcgtg
     3361 gtatcaccgg ttggttccac tctctgttgc gggcaacttc agcagcacgt aggggaagct
     3421 tttattacgc tgcaagggcg taattttcgt cgttcgctgc ggaggtcacc gccatcgaca
     3481 gcggtacgcc ctgcagccat tccgggtggt agcgcagcag catctccagg ccgcgctcct
     3541 ggtaggccgg gaagctgatc gcgaccagcg tctcgccctt gaccttctgc ggcgggccga
     3601 ggcgctggag gatcacgccg ttgcgaacga aatagcgctc catggccgtg gtggtcaccg
     3661 ccaccaccga actggcgccc aggtaccagg cgcattgcag gctggaccag aatatcttca
     3721 tcgccagctg cggatcgtcc gccgcgctgg cggcgtagcg cgaaagctcc cagaccgacg
     3781 gatcgctcgg cggggtttcg ctgcacaggt aggcgaagac gtccttgagc aggtaggcgt
     3841 cggtcgtcgg cagcaggcgg gcgcaaccgc agatgccctg gcggctcatg gcgacgatgt
     3901 agcgggtttg cggatggtcg aactggtcga attcctggtc gcggaccctg gaggtggaga
     3961 ccacgtccca gcccagcttc tcgatgaaga cctgatgccg gtagcgtccc agctcggcga
     4021 tcatggcggc ggaaagccct tccagcgatt cagagagcaa ttcgatcata ctagtatttc
     4081 tccttagcat gattaagatg tttcagtacg aaaattgctt tcattgttga tctccttttt
     4141 aagtggcatg caattgttat ccgctcacaa ttgtgctcat tatcaattgt gagcgctcac
     4201 aagtaaaact gccagatttc acaggatcga cagttcatag gtgattgctc aggacatttc
     4261 tgttagaagg aatcgttttc cttacttttc cttacgcaca agagttccgt agctgttcaa
     4321 gtttgtgttt caactgttct cgtcgtttcc gcaacaagtc ctcttcagaa atgagctttt
     4381 gctcctctgc ttggacggac aggatgtatg ctgtggcttt tttaaggata actaccttgg
     4441 gggccttttc attgttttcc aactccggga tctggtcacg cagggcaaaa aagctccgtt
     4501 ttagctcgtt cctcctctgg cgctccaaga cgttgtgtgt tcgcctcttg acattctcct
     4561 cggtgtccga gggccctgtg tgaaattgtt atccgctcac aattccacac agacgtcgga
     4621 attgccagct ggggcgccct ctggtaaggt tgggaagctc gagaagcgcg tctgaatagg
     4681 cttcttgtat tcctgccaca cagggaaacc atcaggagtt acccaatgca cagcgcaacg
     4741 cttgcgaaga atctctccag tcttcttatc tttgacctca gcagccagca gcttagcagc
     4801 agacttaagc cagttcattg cttcaaccgc agctaccacc gtcacgctca cagattccca
     4861 aatcagctta gccatgtatc cagcagcctg attcggctga gtgaacatca gacccttgcc
     4921 ggaatcaata gctggctgaa tggtatcttc cagcacttgt tgacggaagc cgaactcttt
     4981 ggacccgtaa gccagcgtca tgactgaacg cttagtcaca ctgcgagtaa caccgtaagc
     5041 cagccattga ccagccagtg ccttagtgcc cagcttgact ttctcagaga tttcaccagt
     5101 gttctcatcg gtcacggtaa ctacttcgtt atcggtccca ttgattgcgt ctgcttgtag
     5161 aatctcgttg actttcttag caacaatccc gtagatgtcc tgaacggttt cactaggaag
     5221 caagttaaga cgtctgtgtg gaattgtgag cggataacaa tttcacacag ggccctcgga
     5281 caccgaggag aatgtcaaga ggcgaacaca caacgtcttg gagcgccaga ggaggaacga
     5341 gctaaaacgg agcttttttg ccctgcgtga ccagatcccg gagttggaaa acaatgaaaa
     5401 ggcccccaag gtagttatcc ttaaaaaagc cacagcatac atcctgtccg tccaagcaga
     5461 ggagcaaaag ctcatttctg aagaggactt gttgcggaaa cgacgagaac agttgaaaca
     5521 caaacttgaa cagctacgga actcttgtgc gtaaggaaaa gtaaggaaaa cgattccttc
     5581 taacagaaat gtcctgagca atcacctatg aactgtcgat cctgtgaaat ctggcagttt
     5641 tacttgtgag cgctcacaat tgataatgag cacaattgtg agcggataac aattgcatgc
     5701 cacttaaaaa ggagatcaac aatgaaagca attttcgtac tgaaacatct taatcatgct
     5761 aaggagaaat actagtatgc gtaaaggcga agagctgttc actggtgtcg tccctattct
     5821 ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg
     5881 tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt
     5941 accttggccg actctggtaa cgacgctgac ttggggtgtt cagtgctttg ctcgttatcc
     6001 ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga
     6061 acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga
     6121 aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag aagacggcaa
     6181 tatcctgggc cataagctgg aatacaatta catcagcgac aatgtttaca tcaccgccga
     6241 taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag
     6301 cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct
     6361 gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg
     6421 cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga
     6481 actgtacaaa gcagcgaacg acgaaaatta cgcccttgca gcgtaataa
//
