LOCUS       sequence-112076-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Strong P2 activator plasmid for two-strain system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  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 6013)
  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: pC247
FEATURES             Location/Qualifiers
     source          1..6013
                     /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    3640..3656
                     /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(3840..3869)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    4071..4087
                     /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    4729..4745
                     /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             4947..4976
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    5160..5176
                     /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             5261..5974
                     /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 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 cgctgcggag gtcaccgcca tcgacagcgg tacgccctgc agccattccg ggtggtagcg
     3001 cagcagcatc tccaggccgc gctcctggta ggccgggaag ctgatcgcga ccagcgtctc
     3061 gcccttgacc ttctgcggcg ggccgaggcg ctggaggatc acgccgttgc gaacgaaata
     3121 gcgctccatg gccgtggtgg tcaccgccac caccgaactg gcgcccaggt accaggcgca
     3181 ttgcaggctg gaccagaata tcttcatcgc cagctgcgga tcgtccgccg cgctggcggc
     3241 gtagcgcgaa agctcccaga ccgacggatc gctcggcggg gtttcgctgc acaggtaggc
     3301 gaagacgtcc ttgagcaggt aggcgtcggt cgtcggcagc aggcgggcgc aaccgcagat
     3361 gccctggcgg ctcatggcga cgatgtagcg ggtttgcgga tggtcgaact ggtcgaattc
     3421 ctggtcgcgg accctggagg tggagaccac gtcccagccc agcttctcga tgaagacctg
     3481 atgccggtag cgtcccagct cggcgatcat ggcggcggaa agcccttcca gcgattcaga
     3541 gagcaattcg atcatactag tatttctcct tagcatgatt aagatgtttc agtacgaaaa
     3601 ttgctttcat tgttgatctc ctttttaagt ggcatgcaat tgttatccgc tcacaattgt
     3661 gctcattatc aattgtgagc gctcacaagt aaactgccag atttcacagg atcgacagtt
     3721 cataggtgat tgctcaggac atttctgtta gaaggaatcg ttttccttac ttttccttac
     3781 gcacaagagt tccgtagctg ttcaagtttg tgtttcaact gttctcgtcg tttccgcaac
     3841 aagtcctctt cagaaatgag cttttgctcc tctgcttgga cggacaggat gtatgctgtg
     3901 gcttttttaa ggataactac cttgggggcc ttttcattgt tttccaactc cgggatctgg
     3961 tcacgcaggg caaaaaagct ccgttttagc tcgttcctcc tctggcgctc caagacgttg
     4021 tgtgttcgcc tcttgacatt ctcctcggtg tccgagggcc ctgtgtgaaa ttgttatccg
     4081 ctcacaattc cacacagacg tcggaattgc cagctggggc gccctctggt aaggttggga
     4141 agctcgagaa gcgcgtctga ataggcttct tgtattcctg ccacacaggg aaaccatcag
     4201 gagttaccca atgcacagcg caacgcttgc gaagaatctc tccagtcttc ttatctttga
     4261 cctcagcagc cagcagctta gcagcagact taagccagtt cattgcttca accgcagcta
     4321 ccaccgtcac gctcacagat tcccaaatca gcttagccat gtatccagca gcctgattcg
     4381 gctgagtgaa catcagaccc ttgccggaat caatagctgg ctgaatggta tcttccagca
     4441 cttgttgacg gaagccgaac tctttggacc cgtaagccag cgtcatgact gaacgcttag
     4501 tcacactgcg agtaacaccg taagccagcc attgaccagc cagtgcctta gtgcccagct
     4561 tgactttctc agagatttca ccagtgttct catcggtcac ggtaactact tcgttatcgg
     4621 tcccattgat tgcgtctgct tgtagaatct cgttgacttt cttagcaaca atcccgtaga
     4681 tgtcctgaac ggtttcacta ggaagcaagt taagacgtct gtgtggaatt gtgagcggat
     4741 aacaatttca cacagggccc tcggacaccg aggagaatgt caagaggcga acacacaacg
     4801 tcttggagcg ccagaggagg aacgagctaa aacggagctt ttttgccctg cgtgaccaga
     4861 tcccggagtt ggaaaacaat gaaaaggccc ccaaggtagt tatccttaaa aaagccacag
     4921 catacatcct gtccgtccaa gcagaggagc aaaagctcat ttctgaagag gacttgttgc
     4981 ggaaacgacg agaacagttg aaacacaaac ttgaacagct acggaactct tgtgcgtaag
     5041 gaaaagtaag gaaaacgatt ccttctaaca gaaatgtcct gagcaatcac ctatgaactg
     5101 tcgatcctgt gaaatctggc agtttacttg tgagcgctca caattgataa tgagcacaat
     5161 tgtgagcgga taacaattgc atgccactta aaaaggagat caacaatgaa agcaattttc
     5221 gtactgaaac atcttaatca tgctaaggag aaatactagt atgcgtaaag gcgaagagct
     5281 gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt
     5341 ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat
     5401 ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttgggg
     5461 tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc
     5521 catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa
     5581 aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg
     5641 cattgacttt aaagaagacg gcaatatcct gggccataag ctggaataca attacatcag
     5701 cgacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat
     5761 tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc
     5821 aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct
     5881 gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc
     5941 gggcatcacg catggtatgg atgaactgta caaagcagcg aacgacgaaa attacgccct
     6001 tgcagcgtaa taa
//
