LOCUS       sequence-224207-        3947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PE_4: Prhl_lac_bD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3947)
  AUTHORS   Chen Y, Ho JML, Shis DL, Gupta C, Long J, Wagner DS, Ott W, Josic K,
            Bennett MR
  TITLE     Tuning the dynamic range of bacterial promoters regulated by 
            ligand-inducible transcription factors.
  JOURNAL   Nat Commun. 2018 Jan 4;9(1):64. doi: 10.1038/s41467-017-02473-5.
  PUBMED    29302024
REFERENCE   2  (bases 1 to 3947)
  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"; journalName: "Nat 
            Commun."; date: "2018-01-4"; volume: "9(1):64. doi"; pages: " 
            10.1038/s41467-017-02473-5"
COMMENT     Sequence Label: PE_4: Prhl_lac_bD
FEATURES             Location/Qualifiers
     source          1..3947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..85
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             286..315
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    500..516
                     /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             601..1314
                     /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      1466..1493
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1642..2457)
                     /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="MSHIQRETSCSRPRLNSSMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2365..2384
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2449..2468
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2579..3167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3068..3087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3321..3338
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3353..3492
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3418..3437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3578..3600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3594..3785)
                     /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 tgtggatccc tatcagtgat agagattcac 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 gtctgacatg tgagcgctca
      481 caatttaaat tgagcacaat tgtgagcgga taacaattgc atgccactta aaaaggagat
      541 caacaatgaa agcaattttc gtactgaaac atcttaatca tgctaaggag aaatactagt
      601 atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt
      661 gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt
      721 aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg gccgactctg
      781 gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag
      841 catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt
      901 aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta
      961 aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct gggccataag
     1021 ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc
     1081 attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat
     1141 cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat
     1201 ctgagctacc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg
     1261 ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgatga
     1321 agcttcccct acgtgctgct gaagttgccc gcaacagaga gtggaaccaa ccggtgatac
     1381 cacgatacta tgactgagag tcaacgccat gagcggcctc atttcttatt ctgagttaca
     1441 acagtccgca ccgctgtctc acactggctc accttcgggt gggcctttct gcgtttatat
     1501 actagagaga gaatataaaa agccagatta ttaatccggc ttttttatta tttcctagct
     1561 agccggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1621 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1681 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1741 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1801 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1861 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1921 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1981 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     2041 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     2101 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     2161 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     2221 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2281 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2341 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2401 catgctggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     2461 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     2521 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2581 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2641 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2701 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2761 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2821 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2881 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2941 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3001 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3061 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3121 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3181 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3241 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3301 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3361 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     3421 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     3481 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3541 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3601 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     3661 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     3721 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     3781 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     3841 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     3901 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagctc
//
