LOCUS       sequence-221703-        3968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PE_5: Pxyl_Lac_bD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3968)
  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 3968)
  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_5: Pxyl_Lac_bD
FEATURES             Location/Qualifiers
     source          1..3968
                     /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    521..537
                     /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             622..1335
                     /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      1487..1514
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1663..2478)
                     /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     2386..2405
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2470..2489
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2600..3188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3089..3108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3342..3359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3374..3513
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3439..3458)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3599..3621
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3615..3806)
                     /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 cgaaataaac caaaaatcgt aatcgaaaga taaaaatctg
      481 taactgacat gtgagcgctc acaatttaaa ttgagcacaa ttgtgagcgg ataacaattg
      541 catgccactt aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa catcttaatc
      601 atgctaagga gaaatactag tatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct
      661 attctggtgg aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt
      721 gaaggtgacg caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg
      781 ccggtacctt ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt
      841 tatccggacc atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg
      901 caggaacgca cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa
      961 tttgaaggcg ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagac
     1021 ggcaatatcc tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc
     1081 gccgataaac aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat
     1141 ggcagcgtgc agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt
     1201 ctgctgccag acaatcacta tctgagctac caaagcgttc tgtctaaaga tccgaacgag
     1261 aaacgcgatc atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg
     1321 gatgaactgt acaaatgatg aagcttcccc tacgtgctgc tgaagttgcc cgcaacagag
     1381 agtggaacca accggtgata ccacgatact atgactgaga gtcaacgcca tgagcggcct
     1441 catttcttat tctgagttac aacagtccgc accgctgtct cacactggct caccttcggg
     1501 tgggcctttc tgcgtttata tactagagag agaatataaa aagccagatt attaatccgg
     1561 cttttttatt atttcctagc tagccgggga aatgtgcgcg gaacccctat ttgtttattt
     1621 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     1681 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1741 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1801 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1861 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     1921 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1981 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     2041 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     2101 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2161 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2221 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2281 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2341 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2401 atacccatat aaatcagcat ccatgctgga atttaatcgc ggcctagagc aagacgtttc
     2461 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     2521 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     2761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3241 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3301 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3361 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3421 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3481 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     3541 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3601 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     3661 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     3721 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     3781 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     3841 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     3901 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     3961 cagagctc
//
