LOCUS       sequence-304396-        3148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ClpP protease in the cell-free TX-TL system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3148)
  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 3148)
  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: pBEST_OR2-OR1-Pr-UTR1_ClpP-T500
FEATURES             Location/Qualifiers
     source          1..3148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(836..853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1007..1595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1087..1106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1766..2557,2558..2626))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2389..2408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2627..2731)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2799..2817
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgtc atacagcggc gaacgagata
       61 actttgcacc ccatatggcg ctggtgccga tggtcattga acagacctca cgcggtgagc
      121 gctcttttga tatctattct cgtctactta aggaacgcgt catttttctg actggccagg
      181 ttgaagacca catggctaac ctgattgtgg cgcagatgct gttcctggaa gcggaaaacc
      241 cagaaaaaga tatctatctg tacattaact ccccaggcgg ggtgatcact gccgggatgt
      301 ctatctatga caccatgcag tttatcaagc ctgatgtcag caccatctgt atgggccagg
      361 cggcctcgat gggcgctttc ttgctgaccg caggggcaaa aggtaaacgt ttttgcctgc
      421 cgaattcgcg cgtgatgatt caccaaccgt tgggcggcta ccagggacag gcgaccgata
      481 tcgaaattca tgcccgtgaa attctgaaag ttaaagggcg catgaatgaa cttatggcgc
      541 ttcatacggg tcaatcatta gaacagattg aacgtgatac cgagcgcgat cgcttccttt
      601 ccgcccctga agcggtggaa tacggtctgg tcgattcgat tctgacccat cgtaattgac
      661 tcgagcaaag cccgccgaaa ggcgggcttt tctgtgtcga ccgatgccct tgagagcctt
      721 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
      781 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
      841 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
      901 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      961 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1021 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1081 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1141 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1201 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1261 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1321 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1381 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1441 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1501 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1561 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1621 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1681 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1741 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1861 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1921 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2641 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2701 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     2761 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     2821 ttcgtcttca agaattctgg cgaatcctct gaccagccag aaaacgacct ttctgtggtg
     2881 aaaccggatg ctgcaattca gagcggcagc aagtggggga cagcagaaga cctgaccgcc
     2941 gcagagtgga tgtttgacat ggtgaagact atcgcaccat cagccagaaa accgaatttt
     3001 gctgggtggg ctaacgatat ccgcctgatg cgtgaacgtg acggacgtaa ccaccgcgac
     3061 atgtgtgtgc tgttccgctg ggcatgctga gctaacaccg tgcgtgttga caattttacc
     3121 tctggcggtg ataatggttg cagctagc
//
