LOCUS       sequence-333530-        2805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TetR 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 2805)
  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 2805)
  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-TetR_p15A
FEATURES             Location/Qualifiers
     source          1..2805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      107..652
                     /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     553..572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     769..786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(926..1020)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1051..1845)
                     /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(1162..1181)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1772..1791
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2005..2024
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2005..2023
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2024..2048
                     /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             2064..2084
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2093..2713
                     /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(2166..2185)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2756..2803
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ggccgcacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
       61 gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg
      121 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
      181 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
      241 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
      301 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
      361 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
      421 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
      481 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
      541 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
      601 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
      661 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg
      721 ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg
      781 agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt
      841 tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca
      901 gtatacactc cgctagcgca ctagtattct caccaataaa aaacgcccgg cggcaaccga
      961 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     1021 caagcgagct ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     1081 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     1141 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     1201 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     1261 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc
     1321 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     1381 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     1441 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     1501 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     1561 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     1621 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     1681 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     1741 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     1801 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     1861 tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc
     1921 catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc
     1981 cgacgtccga aatccatggt cacttaatac gactcactat aggggaattg tgagcggata
     2041 acaattcccg actagaaata attttgttta actttaagaa ggaggaggat ccatgtctag
     2101 attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga
     2161 aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg
     2221 gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca
     2281 ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc
     2341 taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac
     2401 acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca
     2461 aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg
     2521 ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac
     2581 tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga
     2641 gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg
     2701 tgaaagtggg tccgcgtaca gcggatcccg ggaattcaga tcttattaaa agcttctagc
     2761 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttggc
//
