LOCUS       sequence-221693-        3979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PE_2: Plas_lac_bD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3979)
  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 3979)
  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_2: Plas_lac_bD
FEATURES             Location/Qualifiers
     source          1..3979
                     /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    532..548
                     /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             633..1346
                     /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      1498..1525
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1674..2489)
                     /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     2397..2416
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2481..2500
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2611..3199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3100..3119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3353..3370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3385..3524
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3450..3469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3610..3632
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3626..3817)
                     /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 cgattcgagc ctagcaaggg tccgggttca ccgaaatcta
      481 tctcatttgc tagtctgaca tgtgagcgct cacaatttaa attgagcaca attgtgagcg
      541 gataacaatt gcatgccact taaaaaggag atcaacaatg aaagcaattt tcgtactgaa
      601 acatcttaat catgctaagg agaaatacta gtatgcgtaa aggcgaagag ctgttcactg
      661 gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc
      721 gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta
      781 ctggtaaact gccggtacct tggccgactc tggtaacgac gctgacttat ggtgttcagt
      841 gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg
      901 aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg
      961 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
     1021 ttaaagaaga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
     1081 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
     1141 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
     1201 atggtcctgt tctgctgcca gacaatcact atctgagcta ccaaagcgtt ctgtctaaag
     1261 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
     1321 cgcatggtat ggatgaactg tacaaatgat gaagcttccc ctacgtgctg ctgaagttgc
     1381 ccgcaacaga gagtggaacc aaccggtgat accacgatac tatgactgag agtcaacgcc
     1441 atgagcggcc tcatttctta ttctgagtta caacagtccg caccgctgtc tcacactggc
     1501 tcaccttcgg gtgggccttt ctgcgtttat atactagaga gagaatataa aaagccagat
     1561 tattaatccg gcttttttat tatttcctag ctagccgggg aaatgtgcgc ggaaccccta
     1621 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     1681 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1741 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1801 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1861 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1921 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     1981 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2041 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2101 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2161 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     2221 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2281 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2341 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2401 cgagcccatt tatacccata taaatcagca tccatgctgg aatttaatcg cggcctagag
     2461 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2521 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3301 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3361 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3421 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3481 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3541 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3601 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3661 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3721 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3781 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3841 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     3901 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     3961 tgcggcggga ccagagctc
//
