LOCUS       sequence-214413-        3969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PE_6: Para_tet_bD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3969)
  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 3969)
  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_6: Para_tet_bD
FEATURES             Location/Qualifiers
     source          1..3969
                     /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    518..536
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             620..1333
                     /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      1485..1512
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1661..2476)
                     /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(1661..2476)
                     /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     2384..2403
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2468..2487
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2598..3186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3087..3106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3340..3357
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3372..3514
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3440..3459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3600..3622
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3616..3807)
                     /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 cgacatagca tttttatcca taagattagc ggatcctacc
      481 tctgacatcc ctatcagtga tagataaatt gagcacatcc ctatcagtga tagagatgca
      541 tgccacttaa aaaggagatc aacaatgaaa gcaattttcg tactgaaaca tcttaatcat
      601 gctaaggaga aatactagta tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat
      661 tctggtggaa ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga
      721 aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc
      781 ggtaccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta
      841 tccggaccat atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca
      901 ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt
      961 tgaaggcgat accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg
     1021 caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc
     1081 cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg
     1141 cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct
     1201 gctgccagac aatcactatc tgagctacca aagcgttctg tctaaagatc cgaacgagaa
     1261 acgcgatcat atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga
     1321 tgaactgtac aaatgatgaa gcttccccta cgtgctgctg aagttgcccg caacagagag
     1381 tggaaccaac cggtgatacc acgatactat gactgagagt caacgccatg agcggcctca
     1441 tttcttattc tgagttacaa cagtccgcac cgctgtctca cactggctca ccttcgggtg
     1501 ggcctttctg cgtttatata ctagagagag aatataaaaa gccagattat taatccggct
     1561 tttttattat ttcctagcta gccggggaaa tgtgcgcgga acccctattt gtttattttt
     1621 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
     1681 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1741 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     1801 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     1861 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     1921 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     1981 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     2041 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     2101 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     2161 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     2221 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     2281 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     2341 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     2401 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     2461 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     2521 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3301 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3361 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3421 atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3481 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3541 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3601 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3661 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3721 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3781 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     3841 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3901 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3961 ccagagctc
//
