LOCUS       sequence-252586-        8871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pseudomonas aeruginosa disaggregase ClpG cloned as ClpG-6xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8871)
  AUTHORS   Lee C, Franke KB, Kamal SM, Kim H, Lunsdorf H, Jager J, Nimtz M, 
            Trcek J, Jansch L, Bukau B, Mogk A, Romling U
  TITLE     Stand-alone ClpG disaggregase confers superior heat tolerance to 
            bacteria.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Jan 9;115(2):E273-E282. doi: 
            10.1073/pnas.1712051115. Epub 2017 Dec 20.
  PUBMED    29263094
REFERENCE   2  (bases 1 to 8871)
  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"; doi: 
            "10.1073/pnas.1712051115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-01-9- 9"; volume: "115"; issue: "2"; pages: "E273-E282"
COMMENT     Sequence Label: pSG019
FEATURES             Location/Qualifiers
     source          1..8871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(260..793)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(982..987,988..1004,1005..1010))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1280..1301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1316..1321,1322..1339,1340..1346)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1354..1370
                     /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     1359..1381
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1378..1394
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1378..1394
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1413..1433
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1415..1433
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1462..1478
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1463..1479
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1571..2449)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2476..2760
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2683..2702
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             5603..5620
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5661..5680)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5662..5680)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5690..5706)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5699..5721)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(6415..7077)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      7078..7849
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
      241 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
      301 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
      361 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
      421 cggtggcaat gccctgcctc cggtgctctc cggagactgc gagatcatag atatagatct
      481 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
      541 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
      601 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
      661 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
      721 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
      781 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
      841 tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa
      901 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
      961 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt
     1021 gccttcatcc gtttccacgg tgtgcgtcca tgggcaaata ttatacgcaa ggcgacaagg
     1081 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     1141 gaatgcttaa tgaattacaa cagtttttat gcatgcgccc aatacgcaaa ccgcctctcc
     1201 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     1261 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     1321 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     1381 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     1441 agctgggtac cgggcccccc ctcgaggtcg acggtatcga tgcataatgt gcctgtcaaa
     1501 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat
     1561 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     1621 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
     1681 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     1741 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     1801 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     1861 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     1921 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     1981 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     2041 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     2101 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     2161 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     2221 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     2281 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
     2341 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
     2401 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
     2461 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
     2521 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     2581 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     2641 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     2701 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     2761 acccgttttt ttgggctagc ccttccgtgg aaacaggagt cctcccatgg cccaggaact
     2821 ttgcgctatc tgccacgaac gtcccgccgt cgcccgcgtc agcctggtac agaacggcca
     2881 gcgccgcgaa ctcgccctct gcgaactgca ctaccgccag ttgatgcgcc agcagcgcat
     2941 gcgctcgccg ctggagtcgc tgttcggcgg cggcagcccg ttcgacgaga tcttctccgg
     3001 cttcggcgag cagagcccgg tcaccccggt gcgcgcccgc gagccggagg cggtggacat
     3061 cgccgagtac ttcagcaagc agaccaccga gtacctgcaa cgcgccgcgc aggtcgccgc
     3121 cgaattcggc aagcgcgaag tggacaccga gcacctgctc tacgccctgg ccgacgccga
     3181 cgtggtgcag gcggtgctca agcagttcgg cctgtcgccg gccgacctca agcagtacat
     3241 cgaggccaac gccgtgcgcg gcgccagcaa gggcgaggcg agcgaggaca tgaccatctc
     3301 gccgcgggtg aagagcgcct tgcagcatgc cttcgccctg tcccgcgaac tcggccacag
     3361 ctatgtcggc cccgagcacc tgttgctcgg cctggcggcg gtgccggaca gcttcgccgg
     3421 gacgctcctg aagaagtacg gcctgaccga gcaggcgcta cggcagaaag cggtcaaggt
     3481 ggtcggcaag ggcgccgagg acggccgcgt ggacggcccg agcaacactc cgcaactgga
     3541 caagttcagc cgcgacctga cccggctggc ccgcgaaggc aagctcgatc cggtgatcgg
     3601 ccgctcgaag gaagtcgaga ccaccatcga ggttctcgcc cggcgcaaga agaacaaccc
     3661 ggtgctgatc ggcgagcccg gcgtcggcaa gaccgccatc gtcgaaggcc tggcccagcg
     3721 catggtccag ggcgaggtgc cggaggtgct gcgcgacaag cgcctggtcg aactgaacat
     3781 caacgccatg gtcgccggcg ccaagtaccg cggcgagttc gaggaacgcc tcaagcaggt
     3841 gatggacgaa ctgcaggcgg cgcagagcga gatcatcctg ttcatcgacg aggtgcacac
     3901 catcgtcggt gccggccagg gcggcggcga aggcgggctg gacgtggcca acgtgctgaa
     3961 gccggcgatg gcgcgtggcg agatgaacct gatcggcgcc accaccctca acgagtacca
     4021 gaagtacatc gagaaggacg ccgcgctgga gcggcgcttc cagccggtgt tcgtccccga
     4081 gccgacggtg gagcagacca tctccatcct ccgtggcctg cgcgacaagc tcgaaggcca
     4141 ccacaaggtg accatccgcg acgaggcctt cgtcgccgcc gccgagctgt ccgatcgcta
     4201 catcggcaac cgcttcctgc cggacaaggc catcgacctg atcgaccagg ccgccgcccg
     4261 cgtgcgcatc gccagcacct cgcggccggc cgagatccag gaactggagg ccgagctggc
     4321 gcagctcaag cgcgagcagg actacgccgc cagccgcaag tggtacgacg aggcgaaggt
     4381 cttcgagaaa cgcatccagg aacgcaagga acacctcgag cagatcaccg agcgctggca
     4441 gcagacccag gggtcgaaga ccgaggaggt gcgggtcgag gacatcgccg agatcatctc
     4501 caggctcacc ggcatcccgg tcaccgaact gaccgccgag gagcgcgaga agctcctgca
     4561 gatggaagag cgcctgcacc agcgggtgat cggccagcag gaagcgatca ccgcggtcag
     4621 cgacgccgtg cgcctggccc gcgccgggct gcgccagggc agccggccga tcgccacctt
     4681 cctgttcctc ggcccgaccg gggtgggcaa gaccgagctg gccaaggccc tggccgaggt
     4741 ggtgttcggc gacgaagacg cgatgatccg catcgacatg agcgaataca tggagcgcca
     4801 cgcggtgtcc cggctgatcg gtgcgccgcc gggctacgtc ggctacgacg agggcggcca
     4861 gctcaccgag cgggtgcgcc ggcggccgta cagcgtgatc ctgctcgatg agatcgagaa
     4921 ggcccacgcc gacgtcaaca acatcctcct gcaggtgttc gacgacggcc gcctgaccga
     4981 cggcaagggg cgcgtggtgg acttcaccaa taccatcatc atcgccacca gcaacctcgg
     5041 ctccgagctg atcatgaaga acgcccaggc cggcgagttc gcccagccgc cggagaagct
     5101 caagcgcgaa ctgatgacta ccctgcgcgg acatttccgc ccggaattcc tcaaccgcct
     5161 cgacgaggtg atcgtcttcg agtcgctgag caaggcgcag atcgaggaca tcgtgcgcct
     5221 gcaactggag cgggtgaagc gcgccgcgca cgctcaggac atctacctgc acatcgacga
     5281 cagcctggtc ggccacctcg ccgaagaggc ctaccagccg gagttcggcg cccgcgagct
     5341 gaagcggcag atccgccagc aactggagac gcgcctggcc acggcgatgc tcaagggcga
     5401 ggtgaaagag ggcgagacgg tcaccttctt ctacgacgcc aaggacggcg tcggctaccg
     5461 caagggcgcc gcgccgaagc cggcggcacg caagaaatcc ggcgccggcg agacgcccaa
     5521 gggccgcgcg acggcggcga ggaagccggc ggcgaagaaa ggcgccgccg ccaagggcaa
     5581 ggccgacaag ccgaaggcca agcatcatca tcatcatcac tgatctagag cggccgccac
     5641 cgcggtggag ctccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     5701 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5761 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5821 agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc
     5881 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgtactgc gatgagtggc
     5941 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     6001 cctgaataag tgataataag cggatgaatg gcagaaattc gaaagcaaat tcgacccggt
     6061 cgtcggttca gggcagggtc gttaaatagc cgcttatgtc tattgctggt ttaccggttt
     6121 attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc agtttgctca
     6181 ggctctcccc gtggaggtaa taattgacga tatgatcatt tattctgcct cccagagcct
     6241 gataaaaacg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     6301 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgaggcgg
     6361 ctacagccga tagtctggaa cagcgcactt acgggttgct gcgcaaccca agtgctaccg
     6421 gcgcggcagc gtgacccgtg tcggcggctc caacggctcg ccatcgtcca gaaaacacgg
     6481 ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc cattcctccg tttcggtcaa
     6541 ggctggcagg tctggttcca tgcccggaat gccgggctgg ctgggcggct cctcgccggg
     6601 gccggtcggt agttgctgct cgcccggata cagggtcggg atgcggcgca ggtcgccatg
     6661 ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc acggcggcac tgaacaccga
     6721 caggcgcaac tggtcgcggg gctggcccca cgccacgcgg tcattgacca cgtaggccga
     6781 cacggtgccg gggccgttga gcttcacgac ggagatccag cgctcggcca ccaagtcctt
     6841 gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc ttggaaagtg tcttctggct
     6901 gaccaccacg gcgttctggt ggcccatctg cgccacgagg tgatgcagca gcattgccgc
     6961 cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc gctttgcgtt ccgtttgcac
     7021 ccagtgaccg ggcttgttct tggcttgaat gccgatttct ctggactgcg tggccatgct
     7081 tatctccatg cggtaggggt gccgcacggt tgcggcacca tgcgcaatca gctgcaactt
     7141 ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta cagattttct
     7201 ttaacctacg caatgagcta ttgcgggggg tgccgcaatg agctgttgcg tacccccctt
     7261 ttttaagttg ttgattttta agtctttcgc atttcgccct atatctagtt ctttggtgcc
     7321 caaagaaggg cacccctgcg gggttccccc acgccttcgg cgcggctccc cctccggcaa
     7381 aaagtggccc ctccggggct tgttgatcga ctgcgcggcc ttcggccttg cccaaggtgg
     7441 cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc tcggggggca ggcgggcggg
     7501 cttcgccctt cgactgcccc cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc
     7561 aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg tccaaccggc
     7621 aagcgaagcg cgcaggccgc aggccggagg cttttcccca gagaaaatta aaaaaattga
     7681 tggggcaagg ccgcaggccg cgcagttgga gccggtgggt atgtggtcga aggctgggta
     7741 gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt
     7801 ccagcagggt tgtccacggg ccgagcgaag cgagccagcc ggtggccgct cgcggccatc
     7861 gtccacatat ccacgggctg gcaagggagc gcagcgaccg cgcagggcga agcccggaga
     7921 gcaagcccgt agggcgccgc agccgccgta ggcggtcacg actttgcgaa gcaaagtcta
     7981 gtgagtatac tcaagcattg agtggcccgc cggaggcacc gccttgcgct gcccccgtcg
     8041 agccggttgg acaccaaaag ggaggggcag gcatggcggc atacgcgatc atgcgatgca
     8101 agaagctggc gaaaatgggc aacgtggcgg ccagtctcaa gcacgcctac cgcgagcgcg
     8161 agacgcccaa cgctgacgcc agcaggacgc cagagaacga gcactgggcg gccagcagca
     8221 ccgatgaagc gatgggccga ctgcgcgagt tgctgccaga gaagcggcgc aaggacgctg
     8281 tgttggcggt cgagtacgtc atgacggcca gcccggaatg gtggaagtcg gccagccaag
     8341 aacagcaggc ggcgttcttc gagaaggcgc acaagtggct ggcggacaag tacggggcgg
     8401 atcgcatcgt gacggccagc atccaccgtg acgaaaccag cccgcacatg accgcgttcg
     8461 tggtgccgct gacgcaggac ggcaggctgt cggccaagga gttcatcggc aacaaagcgc
     8521 agatgacccg cgaccagacc acgtttgcgg ccgctgtggc cgatctaggg ctgcaacggg
     8581 gcatcgaggg cagcaaggca cgtcacacgc gcattcaggc gttctacgag gccctggagc
     8641 ggccaccagt gggccacgtc accatcagcc cgcaagcggt cgagccacgc gcctatgcac
     8701 cgcagggatt ggccgaaaag ctgggaatct caaagcgcgt tgagacgccg gaagccgtgg
     8761 ccgaccggct gacaaaagcg gttcggcagg ggtatgagcc tgccctacag gccgccgcag
     8821 gagcgcgtga gatgcgcaag aaggccgatc aagcccaaga gacggcccga g
//
