LOCUS       sequence-342537-        3995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses QF and/or TetR in BL21(DE3) E. coli according to the 
            presence of aTc. There is no T7 terminator sequence between the QF- 
            and TetR-containing circuits. Contains a p15A origin of 
            replication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3995)
  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 3995)
  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-LacO-QF-T7-TetO-TetR_p15A
FEATURES             Location/Qualifiers
     source          1..3995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..46
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    47..71
                     /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             87..107
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        1202..1220
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1221..1239
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1261..1281
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1290..1910
                     /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(1363..1382)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1953..2000
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2109..2654
                     /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     2555..2574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2771..2788
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2934..3028)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3059..3853)
                     /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(3170..3189)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3780..3799
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gaattccgct agccgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
       61 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggaggag tacatatgcc
      121 acccaagcgc aaaacgctta acgctgcggc tgaggctaac gctcatgccg acggacacgc
      181 cgacggaaac gccgacggac acgtggccaa tacggccgcg tcctcgaata atgcgaggtt
      241 cgctgatctc actaacatcg atactccggg tctgggaccc acaactacga ccctgctcgt
      301 ggaaccagca cgctcaaagc gtcaacgagt gtcccgcgca tgcgaccagt gccgtgcagc
      361 ccgagagaaa tgcgacggaa tacagcctgc gtgtttcccg tgcgtttccc agggaaggtc
      421 ctgcacttat caggcttcgc cgaaaaagag gggagttcaa accggttata ttcgtacgct
      481 ggagctcgcc ctcgcctgga tgtttgaaaa tgtcgcgcgt tccgaagatg ccttgcataa
      541 cctcctcgtc cgtgacgccg gacaaggatc agctctgctc gttggtaaag attcgccggc
      601 tgccgagcga ctccatgccc gttgggctac tagccgtgtc aataagagca ttacccgcct
      661 cctccgtcag ttggagctcc ctcctaccgc cacggctacg gcctcgataa tgccgcacgt
      721 gatggagcag cctctcagta ccagcattaa ccccgtcaac gaccgcttca acggtattcc
      781 caaccccact ccgtataact ccgatgcagc tctcgatgct atcactcaga ccaacgatta
      841 tggaagcgta aatacacatg gtatcctctc tacttacccg ccaccggcta cgcaccttaa
      901 tgaagcttcc gtcgctctcg ctcccggtgg cgcccccccc cgaccgcctc ctccgtatgt
      961 tgacagcacg accaatcacc cgccgtacca ctcgaatctg gttccaatgg cgaactttgg
     1021 ttactcgacc gttgattacg atgccatggt tgacgatttg gctagcattg aatacacgga
     1081 cgctgtggat gtcgacccac agtttatgac caatctggga ttcgttcctg gatgtaactt
     1141 ctccgacatt aatacatacg aacagtgata taaccccggt tgctgaggga atagtctcga
     1201 gtaatacgac tcactatagg tccctatcag tgatagagat gcgcaagcct agaaataatt
     1261 ttgtttaact ttaagaagga ggaggatcca tgtctagatt agataaaagt aaagtgatta
     1321 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     1381 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     1441 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     1501 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     1561 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     1621 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     1681 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     1741 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     1801 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     1861 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtcc gcgtacagcg
     1921 gatcccggga attcagatct tattaaaagc ttctagcata accccttggg gcctctaaac
     1981 gggtcttgag gggttttttg gcggccgcac aacttatatc gtatggggct gacttcaggt
     2041 gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga
     2101 tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc
     2161 gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     2221 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     2281 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     2341 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     2401 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     2461 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     2521 caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc
     2581 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     2641 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     2701 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     2761 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg
     2821 ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca
     2881 tcagtgccaa catagtaagc cagtatacac tccgctagcg cactagtgct tggattctca
     2941 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     3001 gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctc
     3061 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     3121 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     3181 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     3241 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     3301 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     3361 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     3421 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     3481 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     3541 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     3601 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     3661 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     3721 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     3781 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     3841 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc
     3901 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     3961 taccagaggg cgccccagct ggcaattccg acgtc
//
