LOCUS       sequence-112121-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P1 activator plasmid for two-strain system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  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 5977)
  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: pC165
FEATURES             Location/Qualifiers
     source          1..5977
                     /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)."
     protein_bind    3671..3687
                     /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(3822..3851)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    4053..4069
                     /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    4711..4727
                     /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             4929..4958
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    5093..5109
                     /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    5124..5140
                     /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             5225..5938
                     /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 gctcagtatc ttgttatccg ctcacaatgt caatcgacag ttcataggtg attgctcagg
     3721 acatttctgt tagaaggaat cgttttcctt acttttcctt acgcacaaga gttccgtagc
     3781 tgttcaagtt tgtgtttcaa ctgttctcgt cgtttccgca acaagtcctc ttcagaaatg
     3841 agcttttgct cctctgcttg gacggacagg atgtatgctg tggctttttt aaggataact
     3901 accttggggg ccttttcatt gttttccaac tccgggatct ggtcacgcag ggcaaaaaag
     3961 ctccgtttta gctcgttcct cctctggcgc tccaagacgt tgtgtgttcg cctcttgaca
     4021 ttctcctcgg tgtccgaggg ccctgtgtga aattgttatc cgctcacaat tccacacaga
     4081 cgtcggaatt gccagctggg gcgccctctg gtaaggttgg gaagctcgag aagcgcgtct
     4141 gaataggctt cttgtattcc tgccacacag ggaaaccatc aggagttacc caatgcacag
     4201 cgcaacgctt gcgaagaatc tctccagtct tcttatcttt gacctcagca gccagcagct
     4261 tagcagcaga cttaagccag ttcattgctt caaccgcagc taccaccgtc acgctcacag
     4321 attcccaaat cagcttagcc atgtatccag cagcctgatt cggctgagtg aacatcagac
     4381 ccttgccgga atcaatagct ggctgaatgg tatcttccag cacttgttga cggaagccga
     4441 actctttgga cccgtaagcc agcgtcatga ctgaacgctt agtcacactg cgagtaacac
     4501 cgtaagccag ccattgacca gccagtgcct tagtgcccag cttgactttc tcagagattt
     4561 caccagtgtt ctcatcggtc acggtaacta cttcgttatc ggtcccattg attgcgtctg
     4621 cttgtagaat ctcgttgact ttcttagcaa caatcccgta gatgtcctga acggtttcac
     4681 taggaagcaa gttaagacgt ctgtgtggaa ttgtgagcgg ataacaattt cacacagggc
     4741 cctcggacac cgaggagaat gtcaagaggc gaacacacaa cgtcttggag cgccagagga
     4801 ggaacgagct aaaacggagc ttttttgccc tgcgtgacca gatcccggag ttggaaaaca
     4861 atgaaaaggc ccccaaggta gttatcctta aaaaagccac agcatacatc ctgtccgtcc
     4921 aagcagagga gcaaaagctc atttctgaag aggacttgtt gcggaaacga cgagaacagt
     4981 tgaaacacaa acttgaacag ctacggaact cttgtgcgta aggaaaagta aggaaaacga
     5041 ttccttctaa cagaaatgtc ctgagcaatc acctatgaac tgtcgattga cattgtgagc
     5101 ggataacaag atactgagca caattgtgag cggataacaa ttgcatgcca cttaaaaagg
     5161 agatcaacaa tgaaagcaat tttcgtactg aaacatctta atcatgctaa ggagaaatac
     5221 tagtatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga
     5281 tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa
     5341 tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac
     5401 tctggtaacg acgctgactt ggggtgttca gtgctttgct cgttatccgg accatatgaa
     5461 gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc
     5521 ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct
     5581 ggtaaaccgc attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca
     5641 taagctggaa tacaattaca tcagcgacaa tgtttacatc accgccgata aacaaaaaaa
     5701 tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc
     5761 tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca
     5821 ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt
     5881 tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaagc
     5941 agcgaacgac gaaaattacg cccttgcagc gtaataa
//
