LOCUS       sequence-116881-        4880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTrc plasmid expressing clpP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4880)
  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 4880)
  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: pKS044
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(144..149,150..166,167..173)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    214..283
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             334..360
                     /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      1185..1271
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1363..1390
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1409..1500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1501..1569,1570..2361)
                     /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(1719..1738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2532..3120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3021..3040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3274..3291
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3306..3446
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3372..3391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3532..3554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3632..3709
                     /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             3710..4792
                     /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(3729..3748)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg
       61 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
      121 ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcc ggctcgtata atgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagcgccgc tgagaaaaag cgaagcggca
      241 ctgctcttta acaatttatc agacaatctg tgtgggcact cgaccggaat tatcgattaa
      301 ctttattatt aaaaattaaa gaggtatata ttaatgtatc gattaaataa ggaggaataa
      361 accatggatc cagaggatgt catacagcgg cgaacgagat aactttgcac cccatatggc
      421 gctggtgccg atggtcattg aacagacctc acgcggtgag cgctcttttg atatctattc
      481 tcgtctactt aaggaacgcg tcatttttct gactggccag gttgaagacc acatggctaa
      541 cctgattgtg gcgcagatgc tgttcctgga agcggaaaac ccagaaaaag atatctatct
      601 gtacattaac tccccaggcg gggtgatcac tgccgggatg tctatctatg acaccatgca
      661 gtttatcaag cctgatgtca gcaccatctg tatgggccag gcggcctcga tgggcgcttt
      721 cttgctgacc gcaggggcaa aaggtaaacg tttttgcctg ccgaattcgc gcgtgatgat
      781 tcaccaaccg ttgggcggct accagggcca ggcgaccgat atcgaaattc atgcccgtga
      841 aattctgaaa gttaaagggc gcatgaatga acttatggcg cttcatacgg gtcaatcatt
      901 agaacagatt gaacgtgata ccgagcgcga tcgcttcctt tccgcccctg aagcggtgga
      961 atacggtctg gtcgattcga ttctgaccca tcgtaattga gagctcgaga tctgcagctg
     1021 gtaccatatg ggaattcgaa gctttctaga actagtacag aatttgcctg gcggcagtag
     1081 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     1141 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     1201 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     1261 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     1321 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     1381 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     1441 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1501 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1561 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1621 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1681 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1741 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1801 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1861 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1921 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1981 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2041 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2101 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2161 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2221 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2281 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2341 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2401 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2461 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2521 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2581 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2641 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2701 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2761 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2821 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2881 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2941 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3001 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3061 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3121 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3181 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3241 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3301 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3361 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3421 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3481 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3541 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc
     3601 gcgcgaaggc gaagcggcat gcatttacgt tgacaccatc gaatggtgca aaacctttcg
     3661 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     3721 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     3781 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     3841 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     3901 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     3961 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     4021 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     4081 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     4141 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     4201 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     4261 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     4321 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     4381 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     4441 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     4501 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     4561 gccgtcaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     4621 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     4681 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4741 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4801 caattaatgt gagttagcgc gaattgatct ggtttgacag cttatcatcg actgcacggt
     4861 gcaccaatgc ttctggcgtc
//
