LOCUS       sequence-222901-        3931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CH110: Plas_lac_bD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3931)
  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 3931)
  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: CH110: Plas_lac_bD
FEATURES             Location/Qualifiers
     source          1..3931
                     /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    469..485
                     /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             582..1295
                     /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      1447..1474
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1623..2438)
                     /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(1623..2438)
                     /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     2346..2365
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2430..2449
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2560..3148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3049..3068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3302..3319
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3334..3476
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3402..3421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3562..3584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3578..3769)
                     /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 cgaatctatc tcatttgcta gtctgacatt gtgagcggat
      481 aacaatactg tttcttcatg catgccactt aaaaaggaga tcaacaatga aagcaatttt
      541 cgtactgaaa catcttaatc atgctaagga gaaatactag tatgcgtaaa ggcgaagagc
      601 tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac ggtcataagt
      661 tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg ctgaagttca
      721 tctgtactac tggtaaactg ccggtacctt ggccgactct ggtaacgacg ctgacttatg
      781 gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc ttcaagtccg
      841 ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac ggcacgtaca
      901 aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt gagctgaaag
      961 gcattgactt taaagaagac ggcaatatcc tgggccataa gctggaatac aattttaaca
     1021 gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg aattttaaaa
     1081 ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag caaaacactc
     1141 caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagctac caaagcgttc
     1201 tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc gtaaccgcag
     1261 cgggcatcac gcatggtatg gatgaactgt acaaatgatg aagcttcccc tacgtgctgc
     1321 tgaagttgcc cgcaacagag agtggaacca accggtgata ccacgatact atgactgaga
     1381 gtcaacgcca tgagcggcct catttcttat tctgagttac aacagtccgc accgctgtct
     1441 cacactggct caccttcggg tgggcctttc tgcgtttata tactagagag agaatataaa
     1501 aagccagatt attaatccgg cttttttatt atttcctagc tagccgggga aatgtgcgcg
     1561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     1621 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1681 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1741 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1801 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1861 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1921 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1981 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2041 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2101 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     2161 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2221 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2281 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2341 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2401 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     2461 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     2521 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2581 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2641 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2701 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2761 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2821 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2881 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2941 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3001 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3061 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3121 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3181 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3241 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3301 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3361 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     3421 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     3481 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3541 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3601 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3661 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3721 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     3781 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     3841 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     3901 tggcggtatg gatgcggcgg gaccagagct c
//
