LOCUS       sequence-111018-        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTrc plasmid expressing clpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  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 6534)
  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: pKS042
FEATURES             Location/Qualifiers
     source          1..6534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(148..153,154..170,171..177)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    185..201
                     /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     190..212
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    218..287
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             338..364
                     /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      2842..2928
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3020..3047
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3067..3158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3159..3227,3228..4019)
                     /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(3377..3396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4190..4778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4679..4698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4932..4949
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4964..5104
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5030..5049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5190..5212
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5290..5367
                     /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             5368..6450
                     /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(5387..5406)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt
       61 cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg
      121 ttctggcaaa tattctgaaa tgagctgttg acaattaatc atccggctcg tataatgtgt
      181 ggaattgtga gcggataaca atttcacaca ggaaacagcg ccgctgagaa aaagcgaagc
      241 ggcactgctc tttaacaatt tatcagacaa tctgtgtggg cactcgaccg gaattatcga
      301 ttaactttat tattaaaaat taaagaggta tatattaatg tatcgattaa ataaggagga
      361 ataaaccatg gatccagagg atgctcaatc aagaactgga actcagttta aatatggctt
      421 tcgccagagc gcgcgagcac cgtcatgagt ttatgaccgt cgagcacttg ttactggcgc
      481 tgctcagtaa cccatctgcc cgggaggcgc tggaagcgtg ttctgtggat ttggttgcgc
      541 tccgtcagga actggaagcc tttattgaac aaaccacacc cgttctgcct gccagtgaag
      601 aggagcgcga cacacagccg acgctgagtt ttcagcgtgt actgcaacgt gcggtcttcc
      661 atgtccagtc ctccggtcgc aatgaggtta ccggtgcaaa cgttctggtc gctatcttta
      721 gcgaacagga gtcgcaggcg gcatatctgt tgcgtaaaca tgaagtcagc cgtctcgatg
      781 tggtgaactt tatctctcat ggcacgcgta aagacgagcc gacacagtct tctgatcctg
      841 gcagccagcc aaacagcgaa gaacaagctg gtggggagga acgtatggag aatttcacga
      901 cgaacctgaa tcagcttgcg cgcgtgggcg gaatcgaccc actgattggt cgtgagaagg
      961 agctggagcg tgctattcag gttctctgcc gtcgccgtaa aaacaacccg ctgctggtgg
     1021 gggaatctgg tgtcggtaaa accgcgattg cggaaggtct tgcctggcga attgttcagg
     1081 gcgatgtgcc ggaagtgatg gctgactgta cgatttactc tctcgatatc ggttctctgt
     1141 tagcgggcac aaaatatcgc ggcgactttg aaaaacgttt taaagcgttg ctcaagcagc
     1201 tggagcagga cactaacagc atcctgttta ttgatgagat ccacaccatt atcggtgcgg
     1261 gtgcagcgtc tggtggtcag gtcgatgcgg ctaacctaat caaaccgttg ctctccagcg
     1321 gtaaaattcg tgtaattggt tcgacaacct atcaggagtt cagcaacatt ttcgagaaag
     1381 accgtgctct ggcgcgtcgc ttccagaaaa ttgatattac tgaaccgtcg atcgaagaaa
     1441 ctgttcaaat catcaatggc ctgaaaccga agtatgaagc gcaccacgac gtgcgttata
     1501 ccgcaaaagc ggtgcgtgcg gcggtagagc tggcggtgaa atacattaac gatcgtcatc
     1561 tgccggataa agccattgat gttatcgacg aagcgggcgc tcgcgcacgc ctgatgccgg
     1621 taagcaaacg caagaaaacc gttaatgtgg cggatattga gtccgtggtg gcccgtattg
     1681 cacgcattcc agagaagagt gtttctcaga gtgatcgtga taccctgaaa aacctcggcg
     1741 atcgcttgaa aatgctggtc ttcggtcagg ataaagccat tgaggcgctg actgaagcca
     1801 ttaagatggc gcgtgcaggt ttaggtcacg aacataaacc ggttggttcg ttcctgtttg
     1861 ccggccctac cggggtcggg aaaacagagg tgacggtaca gctttcgaaa gctttgggca
     1921 ttgagcttct gcgctttgat atgtccgagt atatggaacg ccataccgtc agccgtctta
     1981 ttggtgcgcc tccgggatac gttggttttg atcagggcgg tttgctgact gatgcggtca
     2041 tcaagcatcc acatgcggtg ctgctgctgg acgaaatcga gaaagcgcac ccggacgtgt
     2101 tcaatattct gttgcaggtg atggataacg gtacgctgac cgataacaac ggacgcaaag
     2161 cagacttccg taacgtggtg ctggtgatga ccaccaacgc cggggtacgg gaaactgagc
     2221 gcaaatccat tggtcttatc caccaggata acagcaccga tgcgatggaa gagatcaaga
     2281 agatctttac accggaattc cgtaaccgtc tcgacaacat tatctggttt gatcatctgt
     2341 caaccgacgt gatccatcag gtggtggata aattcatcgt cgagttgcag gttcagctgg
     2401 atcagaaagg tgtttctctg gaagtgagcc aggaagcgcg taactggctg gccgagaaag
     2461 gttacgaccg ggcaatgggc gctcgtccga tggcgcgtgt catccaggac aacctgaaaa
     2521 aaccgctcgc caacgaactg ctgtttggtt cgctggtgga cggcggtcag gtcaccgtcg
     2581 cgctggataa agagaaaaat gagctgactt acggattcca gagtgcacaa aagcacaagg
     2641 cggaagcagc gcattaagag ctcgagatct gcagctggta ccatatggga attcgaagct
     2701 ttctagaact agtacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     2761 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     2821 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2881 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     2941 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     3001 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     3061 actctttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3121 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3181 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3241 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3301 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3361 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     3421 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3481 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3541 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3601 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3661 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3721 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3781 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3841 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3901 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3961 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4021 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4081 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4141 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4201 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4261 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4321 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4381 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4441 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4501 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4561 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4621 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4681 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4741 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4801 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4861 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4921 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     4981 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     5041 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     5101 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5161 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5221 gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc
     5281 atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
     5341 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     5401 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     5461 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     5521 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     5581 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     5641 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     5701 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     5761 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     5821 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     5881 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     5941 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     6001 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     6061 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     6121 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     6181 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgtcaaccac catcaaacag
     6241 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     6301 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     6361 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6421 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagcgcga
     6481 attgatctgg tttgacagct tatcatcgac tgcacggtgc accaatgctt ctgg
//
