LOCUS       sequence-131947-        3949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prhl/lac-s reporter plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3949)
  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 3949)
  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: pCH194
FEATURES             Location/Qualifiers
     source          1..3949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    68..84
                     /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)."
     primer_bind     73..95
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             286..315
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    499..515
                     /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             600..1313
                     /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"
     terminator      1465..1492
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1641..2456)
                     /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             complement(1641..2456)
                     /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     2364..2383
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2448..2467
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2578..3166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3067..3086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3320..3337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3352..3494
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3420..3439)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3580..3602
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3596..3787)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agtgcgctcg agcttcccaa ccttaccaga gggcgcccca gctggcaatt ccgacgtctg
       61 tgtggaattg tgagcggata acaatttcac acagggccct cggacaccga ggagaatgtc
      121 aagaggcgaa cacacaacgt cttggagcgc cagaggagga acgagctaaa acggagcttt
      181 tttgccctgc gtgaccagat cccggagttg gaaaacaatg aaaaggcccc caaggtagtt
      241 atccttaaaa aagccacagc atacatcctg tccgtccaag cagaggagca aaagctcatt
      301 tctgaagagg acttgttgcg gaaacgacga gaacagttga aacacaaact tgaacagcta
      361 cggaactctt gtgcgtaagg aaaagtaagg aaaacgattc cttctaacag aaatgtcctg
      421 agcaatcacc tatgaactgt cgatcctgtg aaatctggca gtttacttgt gagcgctcac
      481 aattgataat gagcacaatt gtgagcggat aacaattgca tgccacttaa aaaggagatc
      541 aacaatgaaa gcaattttcg tactgaaaca tcttaatcat gctaaggaga aatactagta
      601 tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg
      661 atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta
      721 aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg ccgactctgg
      781 taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc
      841 atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta
      901 aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa
      961 accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg ggccataagc
     1021 tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca
     1081 ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc
     1141 actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc
     1201 tgagctacca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc
     1261 tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgatgaa
     1321 gcttccccta cgtgctgctg aagttgcccg caacagagag tggaaccaac cggtgatacc
     1381 acgatactat gactgagagt caacgccatg agcggcctca tttcttattc tgagttacaa
     1441 cagtccgcac cgctgtctca cactggctca ccttcgggtg ggcctttctg cgtttatata
     1501 ctagagagag aatataaaaa gccagattat taatccggct tttttattat ttcctagcta
     1561 gccggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1621 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     1681 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     1741 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1801 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1861 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     1921 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     1981 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     2041 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     2101 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     2161 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     2221 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     2281 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     2341 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     2401 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
     2461 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
     2521 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2581 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2641 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2701 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2761 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2821 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2881 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2941 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3001 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3061 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3121 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3181 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3241 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3301 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     3361 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg cactctcagt
     3421 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3481 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3541 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3601 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3661 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3721 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3781 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     3841 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     3901 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagctc
//
