LOCUS       sequence-311106-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses clpX and clpP in E.Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  AUTHORS   Lakshmanan A, Jin Z, Nety SP, Sawyer DP, Lee-Gosselin A, Malounda D,
            Swift MB, Maresca D, Shapiro MG
  TITLE     Acoustic biosensors for ultrasound imaging of enzyme activity.
  JOURNAL   Nat Chem Biol. 2020 Sep;16(9):988-996. doi: 
            10.1038/s41589-020-0591-0. Epub 2020 Jul 13.
  PUBMED    32661379
REFERENCE   2  (bases 1 to 7715)
  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: 
            "10.1038/s41589-020-0591-0"; journalName: "Nat Chem Biol"; date: 
            "2020-09"; volume: "16"; issue: "9"; pages: "988-996"
COMMENT     Sequence Label: araBAD-BCD20-ClpP-BCD17-ClpX
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(84..962)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        989..1273
                     /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     1196..1215
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3659..3677)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3673..3720
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3737..3754)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3907..3993
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4085..4112
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4131..4222
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4680..5135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4767..4786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4977..4998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5691..6350)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWCEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6265..6284
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6351..6453)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6845..6862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6979..7524)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(7059..7078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7600..7619
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg
       61 tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct
      121 tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag
      181 aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg
      241 gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta
      301 atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt
      361 gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc
      421 tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc
      481 agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc
      541 ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg
      601 cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg
      661 cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag
      721 tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt
      781 ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt
      841 cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc
      901 gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc
      961 atacttttca tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat
     1021 tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt
     1081 aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc
     1141 tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc
     1201 atagcatttt tatccataag attagcggat cctacctgac gctttttatc gcaactctct
     1261 actgtttctc catgatactg agcacctctc tactgtttct ccatagggcc caagttcact
     1321 taaaaaggag atcaacaatg aaagcaattt tcgtactgaa acatcttaat catgctgagg
     1381 aaagtttcta atgtcataca gcggcgaacg agataacttt gcaccccata tggcgctggt
     1441 gccgatggtc attgaacaga cctcacgcgg tgagcgctct tttgatatct attctcgtct
     1501 acttaaggaa cgcgtcattt ttctgactgg ccaggttgaa gaccacatgg ctaacctgat
     1561 tgtggcgcag atgctgttcc tggaagcgga aaacccagaa aaagatatct atctgtacat
     1621 taactcccca ggcggggtga tcactgccgg gatgtctatc tatgacacca tgcagtttat
     1681 caagcctgat gtcagcacca tctgtatggg ccaggcggcc tcgatgggcg ctttcttgct
     1741 gaccgcaggg gcaaaaggta aacgtttttg cctgccgaat tcgcgcgtga tgattcacca
     1801 accgttgggc ggctaccagg gacaggcgac cgatatcgaa attcatgccc gtgaaattct
     1861 gaaagttaaa gggcgcatga atgaacttat ggcgcttcat acgggtcaat cattagaaca
     1921 gattgaacgt gataccgagc gcgatcgctt cctttccgcc cctgaagcgg tggaatacgg
     1981 tctggtcgat tcgattctga cccatcgtaa ttgataatac tagatgcatt aataatcaca
     2041 ctggctcacc tctctactgt ttctccatag ggcccaagtt cacttaaaaa ggagatcaac
     2101 aatgaaagca attttcgtac tgaaacatct taatcatgcg gaggagggtt tctaatgaca
     2161 gataaacgca aagatggctc aggcaaattg ctgtattgct ctttttgcgg caaaagccag
     2221 catgaagtgc gcaagctgat tgccggtcca tccgtgtata tctgcgacga atgtgttgat
     2281 ttatgtaacg acatcattcg cgaagagatt aaagaagttg caccgcatcg tgaacgcagt
     2341 gcgctaccga cgccgcatga aattcgcaac cacctggacg attacgttat cggccaggaa
     2401 caggcgaaaa aagtgctggc ggtcgcggta tacaaccatt acaaacgtct gcgcaacggc
     2461 gataccagca atggcgtcga gttgggcaaa agtaacattc tgctgatcgg tccgaccggt
     2521 tccggtaaaa cgctgctggc tgaaacgctg gcgcgcctgc tggatgttcc gttcaccatg
     2581 gccgacgcga ctacactgac cgaagccggt tatgtgggtg aagacgttga aaacatcatt
     2641 cagaagctgt tgcagaaatg cgactacgat gtccagaaag cacagcgtgg tattgtctac
     2701 atcgatgaaa tcgacaagat ttctcgtaag tcagacaacc cgtccattac ccgagacgtt
     2761 tccggtgaag gcgtacagca ggcactgttg aaactgatcg aaggtacggt agctgctgtt
     2821 ccaccgcaag gtgggcgtaa acatccgcag caggaattct tgcaggttga tacctctaag
     2881 atcctgttta tttgtggcgg tgcgtttgcc ggtctggata aagtgatttc ccaccgtgta
     2941 gaaaccggct ccggcattgg ttttggcgcg acggtaaaag cgaagtccga caaagcaagc
     3001 gaaggcgagc tgctggcgca ggttgaaccg gaagatctga tcaagtttgg tcttatccct
     3061 gagtttattg gtcgtctgcc ggttgtcgca acgttgaatg aactgagcga agaagctctg
     3121 attcagatcc tcaaagagcc gaaaaacgcc ctgaccaagc agtatcaggc gctgtttaat
     3181 ctggaaggcg tggatctgga attccgtgac gaggcgctgg atgctatcgc taagaaagcg
     3241 atggcgcgta aaaccggtgc ccgtggcctg cgttccatcg tagaagccgc actgctcgat
     3301 accatgtacg atctgccgtc catggaagac gtcgaaaaag tggttatcga cgagtcggta
     3361 attgatggtc aaagcaaacc gttgctgatt tatggcaagc cggaagcgca acaggcatct
     3421 ggtgaataaa aaacggcggg gtgattgccc cgccgttttt tagtgatgtg atgagatgtg
     3481 cagcttcttt tttcgataca tgagcggagt catcttcatt tcttttcgaa actgattaat
     3541 aaaatagctt gtactattaa accctacttg ataggcgact tccgtcacat ttgcttcttc
     3601 ttgctgcagg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     3661 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     3721 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     3781 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     3841 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     3901 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3961 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4021 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     4081 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     4141 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4201 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4261 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4321 tgctgaagat cagttgggtg cagcaaacta ttaactggcg aactacttac tctagcttcc
     4381 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4441 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4501 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4561 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4621 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4681 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4741 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4801 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4861 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4921 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4981 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     5041 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5101 cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt
     5161 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5221 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5281 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5341 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     5401 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5461 tgctaatcct gttaccagtc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     5521 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     5581 gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt
     5641 caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg
     5701 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     5761 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     5821 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     5881 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     5941 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     6001 agaaactgcc ggaaatcgtc gtggtattca caccagagcg atgaaaacgt ttcagtttgc
     6061 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     6121 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     6181 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     6241 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     6301 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     6361 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     6421 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc
     6481 cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc
     6541 gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt actgatttag
     6601 tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg
     6661 ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctagcg
     6721 gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg
     6781 gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg
     6841 cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt
     6901 acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag
     6961 ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg
     7021 acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7081 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct
     7141 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct
     7201 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta
     7261 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg
     7321 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
     7381 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
     7441 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
     7501 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag
     7561 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     7621 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagatgat
     7681 ctgctcatgt ttgacagctt atcatcgatg cataa
//
