LOCUS       sequence-341154-        3217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses QF activator protein in BL21(DE3) E. coli. Contains p15A 
            origin of replication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3217)
  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 3217)
  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_p15A
FEATURES             Location/Qualifiers
     source          1..3217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..47
                     /label=T7
                     /note="T7 promoter, forward primer"
     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)"
     terminator      1175..1222
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1331..1876
                     /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     1777..1796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1993..2010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2156..2250)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2281..3075)
                     /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(2392..2411)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3002..3021
                     /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 aacagtgaaa gcttctagca taaccccttg gggcctctaa
     1201 acgggtcttg aggggttttt tggcggccgc acaacttata tcgtatgggg ctgacttcag
     1261 gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc tgattaataa
     1321 gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa
     1381 ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta
     1441 actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc
     1501 atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt
     1561 gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac
     1621 tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt
     1681 gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa
     1741 ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag
     1801 tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg
     1861 gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc
     1921 ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga
     1981 acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc
     2041 tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct
     2101 catcagtgcc aacatagtaa gccagtatac actccgctag cgcactagtg cttggattct
     2161 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
     2221 aggtcattac tggatctatc aacaggagtc caagcgagct ctcgaacccc agagtcccgc
     2281 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     2341 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     2401 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     2461 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     2521 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     2581 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     2641 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     2701 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     2761 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     2821 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     2881 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     2941 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     3001 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     3061 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
     3121 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     3181 cttaccagag ggcgccccag ctggcaattc cgacgtc
//
