LOCUS       sequence-116880-        4577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTrc plasmid expressing clpS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4577)
  AUTHORS   Sekar K, Gentile AM, Bostick JW, Tyo KE
  TITLE     N-Terminal-Based Targeted, Inducible Protein Degradation in 
            Escherichia coli.
  JOURNAL   PLoS One. 2016 Feb 22;11(2):e0149746. doi: 
            10.1371/journal.pone.0149746. eCollection 2016.
  PUBMED    26900850
REFERENCE   2  (bases 1 to 4577)
  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: "PLoS One.";
            date: "2016-02-22"; volume: "11(2):e0149746. doi"; pages: " 
            10.1371/journal.pone.0149746. eCollection 2016"
COMMENT     Sequence Label: pKS043
FEATURES             Location/Qualifiers
     source          1..4577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(142..147,148..164,165..171)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    179..195
                     /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)."
     primer_bind     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    212..281
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             332..358
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     terminator      880..966
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1058..1085
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1104..1195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1196..1264,1265..2056)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1414..1433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2227..2815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2716..2735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2969..2986
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3001..3141
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3067..3086)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3227..3249
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3327..3404
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3405..4487
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3424..3443)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc
       61 gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg
      121 caaatattct gaaatgagct gttgacaatt aatcatccgg ctcgtataat gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agcgccgctg agaaaaagcg aagcggcact
      241 gctctttaac aatttatcag acaatctgtg tgggcactcg accggaatta tcgattaact
      301 ttattattaa aaattaaaga ggtatatatt aatgtatcga ttaaataagg aggaataaac
      361 catggatcca gaggatgggt aaaacgaacg actggctgga ctttgatcaa ctggcggaag
      421 aaaaagttcg cgacgcgcta aaaccgccat ctatgtataa agtgatatta gtcaatgatg
      481 attacactcc gatggagttt gttattgacg tgttacaaaa attcttttct tatgatgtag
      541 aacgtgcaac gcaattgatg ctcgctgttc actaccaggg gaaggccatt tgcggagtct
      601 ttaccgccga ggttgcagaa accaaagtgg cgatggtgaa caagtacgcg agggagaatg
      661 agcatccatt gctgtgtacg ctagaaaaag cctgagagct cgagatctgc agctggtacc
      721 atatgggaat tcgaagcttt ctagaactag tacagaattt gcctggcggc agtagcgcgg
      781 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
      841 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
      901 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
      961 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     1021 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     1081 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc
     1141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2341 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2941 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3001 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     3061 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     3121 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3181 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3241 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg
     3301 aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta
     3361 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     3421 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     3481 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     3541 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     3601 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     3661 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     3721 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     3781 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     3841 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     3901 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     3961 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     4021 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     4081 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     4141 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     4201 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     4261 caaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     4321 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     4381 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4441 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4501 aatgtgagtt agcgcgaatt gatctggttt gacagcttat catcgactgc acggtgcacc
     4561 aatgcttctg gcgtcag
//
