LOCUS       sequence-252589-        8914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pseudomonas aeruginosa disaggregase ClpG(GI) cloned as 
            ClpG(GI)-6xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8914)
  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 8914)
  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: pSG020
FEATURES             Location/Qualifiers
     source          1..8914
                     /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"
     regulatory      3028..3037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3841..3850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5646..5663
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5704..5723)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5705..5723)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5733..5749)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5742..5764)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(6458..7120)
                     /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      7121..7892
                     /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 ctttacagga gcatcagcat ggccagaaaa caatgccaag
     2821 tctgcggcca gcccgccacg gtgcgggtgg aagccaatct caatggtcgc cacagcacca
     2881 tgctgttgtg tgacgatcac tatcgccaac tggtgcgcca gcaaaagcgc accgtctcac
     2941 cgctggaagc cttgttcggc tcgcgcagcg ggctgttcga agacttcctt ggcagcgact
     3001 tcttccgcat cggtgacgac gcaccgtcca tggcggccga taccgacgag gtcgtcgatg
     3061 cctcgttcgg cgaacccgcc ccggccggta cgggcaccgc gcgccgtcgc ggcagtgggc
     3121 tcgccagccg tatcagcgaa cagtccgagg ccctattgca ggaggccgcc cgacacgctg
     3181 cagagttcgg gcgcgccgaa gtcgataccg aacacctgct gctggcgcta tccgacagcg
     3241 acgtggtcaa gaccatcctg gggcagttca agatcaaggt cgatgacctc aagcgccaga
     3301 tcgaatccga agccaagcgc ggcgataagc cgttcgaggg cgagatcggc gtgtcgcccc
     3361 gggtcaagga cgcgctcagc cgtgctttcg tggcctccaa cgaactcggc cactcttatg
     3421 tcgggccgga gcatttcctg atcgggctcg ccgaggaagg cgaaggtttg gcggccaacc
     3481 tgctgcgccg ttacggcctc acgccgcaag cgctgcgcca gcaggtaagc aaggtggtcg
     3541 gcaaaggggc cgaggatggc cgcgccgaga cgccgaccaa cacgccggaa ctcgacaagt
     3601 attcgcgcga cctcaccaag atggcgcgcg agggcaagct cgatccggtc atcggccgcg
     3661 cgcaggagat cgagacgacc atcgaagtgc tggcccggcg caagaagaac aacccggtac
     3721 tgatcggcga gcccggcgtc ggcaagaccg ccatcgtcga agggctggcg cagcgcatgg
     3781 tcgccggcga agtgcccgag acgctgcgcg acaagcgtct ggtcgaactc aacatcaacg
     3841 ccatggtggc aggcgccaag taccgcggcg agttcgagga gcgcgtgcag aaggtgctca
     3901 aggaagtgac cgagcaccag ggcgagctga ttctcttcat cgacgaggtg cacaccatcg
     3961 tcggtgcagg ccagggtggc ggcgaaggcg ggctggacgt ggccaacgtg ttcaagccga
     4021 tgatggcgcg cggcgaactg aacctgatcg gcgccaccac gctcaacgag tatcagaagt
     4081 acatcgagaa ggacgccgcg ctggagcgtc gcttccagcc ggtgatggtg cccgagccga
     4141 cggtagcgca gaccatgatg atcctgcgcg gcctgcgtga caccttcgag gcgcaccaca
     4201 aggtcagcat caccgaggat gcgatcatcg ccgccgccga gttgtcggac cgctacatca
     4261 ccgcgcgctt tttgcctgac aaggccatcg acctgctcga ccaggcggcc gcacgcgtga
     4321 agctgtcggc cacggcccgc ccggtggcgg tgcaagagct ggagtccgaa ctgcaccagc
     4381 tgcggcgtga gcaggactat gtggcctcgc gcaagcagta cgacaaggcc gccgagctgg
     4441 gcaagcacat cgaggccaaa gaggccgaac tcaagaagct tgtcgaggaa tgggaacgcg
     4501 agcgcgcctc gggcagtgcc gaagtcaaag ccgagcatgt cgcgcagatc gtctcgcgcc
     4561 tgaccggcat tccggtcaac gagctgacgg tggaagaacg cgagaagctg ctgcatctgg
     4621 agcagcggct gcacgagcgc ctcgtgggcc aggacgaagc ggtgcgcgcg gtggccgatg
     4681 ccgtgcggtt gtcgcgcgcg ggcctgcgcg aaggcagcaa gccggtggcg acttttctgt
     4741 tcctcgggcc gaccggcgtg ggcaagaccg agctcgccaa ggcgctggcc gagtccatct
     4801 atggcgatga aggtgcgctg ctgcgcatcg acatgtccga gtacggggaa cgccataccg
     4861 tggcacgcct ggtgggggcg cctccgggtt acgtcggcta cgacgagggt ggccagctca
     4921 ccgagaaggt gcggcgcaaa ccctacagcg tgttgctgct ggacgagatc gagaaggctc
     4981 accccgacgt ctacaacatc ctgctgcagg tgttcgacga cgggcggctc accgacggca
     5041 agggccgggt ggtggatttc accaatacca tcatcatcgc cacctcgaac ttgggctcgg
     5101 acatcatcca gcgtcggctg aaggcccgtg gcgccgccgg cgaggaatac gagaagacca
     5161 agggcgaggt gatggacgtg ctgcgcggac acttccgccc cgagttcctc aaccgcatcg
     5221 acgagatcat cgtcttccat gcgctgggca aggaggagat ccgccatatc gtcggcctgc
     5281 agctcgatcg tgtggcccgc aacgccgcca gccagggcgt gacgctgacc ttcgatcaga
     5341 ccttgatcga tcacttcgcg gaggaaggct acaagcccga gttcggcgcg cgtgagctca
     5401 agcggctgat ccgcagcgag ttggaaactg ctctggcgcg cgagatgctg ggtggcggta
     5461 tcggcaaggc cgatcacgcc agcgcccgct gggacgacaa ggccgaacgg gtggtcttcg
     5521 agcgccagga gccacccgcg aagccggccg agcctgagaa gcccgatgcc gcgaacgtgg
     5581 ccgagacgcc gccgagcgac gcgagcaagc ctgcgcgcaa gaagaagtca gcgggcggcg
     5641 aatctcacca ccaccaccac cattgatcta gagcggccgc caccgcggtg gagctccaat
     5701 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     5761 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5821 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5881 ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5941 tcagctcatt ttttaaccaa taggccgtac tgcgatgagt ggcagggcgg ggcgtaattt
     6001 ttttaaggca gttattggtg cccttaaacg cctggttgct acgcctgaat aagtgataat
     6061 aagcggatga atggcagaaa ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg
     6121 gtcgttaaat agccgcttat gtctattgct ggtttaccgg tttattgact accggaagca
     6181 gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc tcaggctctc cccgtggagg
     6241 taataattga cgatatgatc atttattctg cctcccagag cctgataaaa acggtgaatc
     6301 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     6361 acgcaacgcg gggaggcaga caaggtatag ggcggcgagg cggctacagc cgatagtctg
     6421 gaacagcgca cttacgggtt gctgcgcaac ccaagtgcta ccggcgcggc agcgtgaccc
     6481 gtgtcggcgg ctccaacggc tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca
     6541 ggcgctgctg cccgcgccgt tcccattcct ccgtttcggt caaggctggc aggtctggtt
     6601 ccatgcccgg aatgccgggc tggctgggcg gctcctcgcc ggggccggtc ggtagttgct
     6661 gctcgcccgg atacagggtc gggatgcggc gcaggtcgcc atgccccaac agcgattcgt
     6721 cctggtcgtc gtgatcaacc accacggcgg cactgaacac cgacaggcgc aactggtcgc
     6781 ggggctggcc ccacgccacg cggtcattga ccacgtaggc cgacacggtg ccggggccgt
     6841 tgagcttcac gacggagatc cagcgctcgg ccaccaagtc cttgactgcg tattggaccg
     6901 tccgcaaaga acgtccgatg agcttggaaa gtgtcttctg gctgaccacc acggcgttct
     6961 ggtggcccat ctgcgccacg aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa
     7021 taagcccggc ccacgcctca tgcgctttgc gttccgtttg cacccagtga ccgggcttgt
     7081 tcttggcttg aatgccgatt tctctggact gcgtggccat gcttatctcc atgcggtagg
     7141 ggtgccgcac ggttgcggca ccatgcgcaa tcagctgcaa cttttcggca gcgcgacaac
     7201 aattatgcgt tgcgtaaaag tggcagtcaa ttacagattt tctttaacct acgcaatgag
     7261 ctattgcggg gggtgccgca atgagctgtt gcgtaccccc cttttttaag ttgttgattt
     7321 ttaagtcttt cgcatttcgc cctatatcta gttctttggt gcccaaagaa gggcacccct
     7381 gcggggttcc cccacgcctt cggcgcggct ccccctccgg caaaaagtgg cccctccggg
     7441 gcttgttgat cgactgcgcg gccttcggcc ttgcccaagg tggcgctgcc cccttggaac
     7501 ccccgcactc gccgccgtga ggctcggggg gcaggcgggc gggcttcgcc cttcgactgc
     7561 ccccactcgc ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg
     7621 ctgcgcgagc cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc
     7681 cgcaggccgg aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg
     7741 ccgcgcagtt ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct
     7801 gtggtcaagc tcgtgggcag gcgcagcctg tccatcagct tgtccagcag ggttgtccac
     7861 gggccgagcg aagcgagcca gccggtggcc gctcgcggcc atcgtccaca tatccacggg
     7921 ctggcaaggg agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc
     7981 cgcagccgcc gtaggcggtc acgactttgc gaagcaaagt ctagtgagta tactcaagca
     8041 ttgagtggcc cgccggaggc accgccttgc gctgcccccg tcgagccggt tggacaccaa
     8101 aagggagggg caggcatggc ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg
     8161 ggcaacgtgg cggccagtct caagcacgcc taccgcgagc gcgagacgcc caacgctgac
     8221 gccagcagga cgccagagaa cgagcactgg gcggccagca gcaccgatga agcgatgggc
     8281 cgactgcgcg agttgctgcc agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac
     8341 gtcatgacgg ccagcccgga atggtggaag tcggccagcc aagaacagca ggcggcgttc
     8401 ttcgagaagg cgcacaagtg gctggcggac aagtacgggg cggatcgcat cgtgacggcc
     8461 agcatccacc gtgacgaaac cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag
     8521 gacggcaggc tgtcggccaa ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag
     8581 accacgtttg cggccgctgt ggccgatcta gggctgcaac ggggcatcga gggcagcaag
     8641 gcacgtcaca cgcgcattca ggcgttctac gaggccctgg agcggccacc agtgggccac
     8701 gtcaccatca gcccgcaagc ggtcgagcca cgcgcctatg caccgcaggg attggccgaa
     8761 aagctgggaa tctcaaagcg cgttgagacg ccggaagccg tggccgaccg gctgacaaaa
     8821 gcggttcggc aggggtatga gcctgcccta caggccgccg caggagcgcg tgagatgcgc
     8881 aagaaggccg atcaagccca agagacggcc cgag
//
