LOCUS       sequence-341152-        2811 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 2811)
  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 2811)
  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..2811
                     /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(932..1026)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1057..1851)
                     /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(1168..1187)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1778..1797
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2011..2030
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2011..2029
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2030..2054
                     /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             2070..2090
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2099..2719
                     /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(2172..2191)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2762..2809
                     /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 ctagtgcttg gattctcacc aataaaaaac gcccggcggc
      961 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     1021 ggagtccaag cgagctctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     1081 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     1141 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     1201 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     1261 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     1321 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     1381 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     1441 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     1501 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     1561 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     1621 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     1681 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     1741 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     1801 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     1861 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     1921 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     1981 caattccgac gtccgaaatc catggtcact taatacgact cactataggg gaattgtgag
     2041 cggataacaa ttcccgacta gaaataattt tgtttaactt taagaaggag gaggatccat
     2101 gtctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     2161 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     2221 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     2281 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     2341 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     2401 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     2461 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac
     2521 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     2581 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     2641 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     2701 taaatgtgaa agtgggtccg cgtacagcgg atcccgggaa ttcagatctt attaaaagct
     2761 tctagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgg c
//
