LOCUS       sequence-341153-        4085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses QF and/or TetR in BL21(DE3) E. coli according to the 
            presence of aTc. Contains a p15A origin of replication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4085)
  AUTHORS   MacDonald IC, Seamons TR, Emmons JC, Javdan SB, Deans TL
  TITLE     Enhanced regulation of prokaryotic gene expression by a eukaryotic 
            transcriptional activator.
  JOURNAL   Nat Commun. 2021 Jul 5;12(1):4109. doi: 10.1038/s41467-021-24434-9.
  PUBMED    34226549
REFERENCE   2  (bases 1 to 4085)
  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: "2021-07-5"; volume: "12(1):4109. doi"; pages: " 
            10.1038/s41467-021-24434-9"
COMMENT     Sequence Label: T7-TetO-QF_T7-LacO-TetR_p15A
FEATURES             Location/Qualifiers
     source          1..4085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..105
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    106..130
                     /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)."
     RBS             146..166
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             175..795
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(248..267)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      838..885
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      994..1539
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1440..1459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1656..1673
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1819..1913)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1944..2738)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2055..2074)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2665..2684
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        2890..2908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2909..2927
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      4036..4083
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ggccgccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
       61 tccgacgtcc gaaatccatg gtcacttaat acgactcact ataggggaat tgtgagcgga
      121 taacaattcc cgactagaaa taattttgtt taactttaag aaggaggagg atccatgtct
      181 agattagata aaagtaaagt gattaacagc gcattagagc tgcttaatga ggtcggaatc
      241 gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc tacattgtat
      301 tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat gttagatagg
      361 caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt acgtaataac
      421 gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt acatttaggt
      481 acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt tttatgccaa
      541 caaggttttt cactagagaa tgcattatat gcactcagcg ctgtggggca ttttacttta
      601 ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga aacacctact
      661 actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca ccaaggtgca
      721 gagccagcct tcttattcgg ccttgaattg atcatatgcg gattagaaaa acaacttaaa
      781 tgtgaaagtg ggtccgcgta cagcggatcc cgggaattca gatcttatta aaagcttcta
      841 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttggcggc cgcacaactt
      901 atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca ctgaaatcta
      961 gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc tgcgcgtaat
     1021 ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc
     1081 taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa aacttgtcct
     1141 ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat caattaccag
     1201 tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga cgatagttac
     1261 cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc agcttggagc
     1321 gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg ccataacagc
     1381 ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga gccgccaggg
     1441 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag
     1501 atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc cgcggccctc
     1561 tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc cgttcgtaag
     1621 ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg
     1681 aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt ctcctgccac
     1741 atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta tacactccgc
     1801 tagcgcacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct
     1861 gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga
     1921 gctctcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
     1981 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     2041 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     2101 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     2161 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc
     2221 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     2281 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     2341 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     2401 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     2461 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     2521 gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
     2581 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     2641 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     2701 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     2761 tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     2821 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccgacgtc
     2881 ccgataaatt aatacgactc actataggtc cctatcagtg atagagagtt ctttgcaggg
     2941 cttcccaacc tttaacttta agaaggagga ggatccatgc cacccaagcg caaaacgctt
     3001 aacgctgcgg ctgaggctaa cgctcatgcc gacggacacg ccgacggaaa cgccgacgga
     3061 cacgtggcca atacggccgc gtcctcgaat aatgcgaggt tcgctgatct cactaacatc
     3121 gatactccgg gtctgggacc cacaactacg accctgctcg tggaaccagc acgctcaaag
     3181 cgtcaacgag tgtcccgcgc atgcgaccag tgccgtgcag cccgagagaa atgcgacgga
     3241 atacagcctg cgtgtttccc gtgcgtttcc cagggaaggt cctgcactta tcaggcttcg
     3301 ccgaaaaaga ggggagttca aaccggttat attcgtacgc tggagctcgc cctcgcctgg
     3361 atgtttgaaa atgtcgcgcg ttccgaagat gccttgcata acctcctcgt ccgtgacgcc
     3421 ggacaaggat cagctctgct cgttggtaaa gattcgccgg ctgccgagcg actccatgcc
     3481 cgttgggcta ctagccgtgt caataagagc attacccgcc tcctccgtca gttggagctc
     3541 cctcctaccg ccacggctac ggcctcgata atgccgcacg tgatggagca gcctctcagt
     3601 accagcatta accccgtcaa cgaccgcttc aacggtattc ccaaccccac tccgtataac
     3661 tccgatgcag ctctcgatgc tatcactcag accaacgatt atggaagcgt aaatacacat
     3721 ggtatcctct ctacttaccc gccaccggct acgcacctta atgaagcttc cgtcgctctc
     3781 gctcccggtg gcgccccccc ccgaccgcct cctccgtatg ttgacagcac gaccaatcac
     3841 ccgccgtacc actcgaatct ggttccaatg gcgaactttg gttactcgac cgttgattac
     3901 gatgccatgg ttgacgattt ggctagcatt gaatacacgg acgctgtgga tgtcgaccca
     3961 cagtttatga ccaatctggg attcgttcct ggatgtaact tctccgacat taatacatac
     4021 gaacagtgaa agcttctagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     4081 ttggc
//
