LOCUS       sequence-310166-        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             192..214
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1021..1038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1192..1780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1272..1291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1951..2742,2743..2811))
                     /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     2574..2593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2812..2916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2984..3002
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaagactatc gcaccatcag ccagaaaacc gaattttgct gggtgggcta acgatatccg
       61 cctgatgcgt gaacgtgacg gacgtaacca ccgcgacatg tgtgtgctgt tccgctgggc
      121 atgctgagct aacaccgtgc gtgttgacaa ttttacctct ggcggtgata atggttgcag
      181 ctagcaataa ttttgtttaa ctttaagaag gagatatacc atgtcataca gcggcgaacg
      241 agataacttt gcaccccata tggcgctggt gccgatggtc attgaacaga cctcacgcgg
      301 tgagcgctct tttgatatct attctcgtct acttaaggaa cgcgtcattt ttctgactgg
      361 ccaggttgaa gaccacatgg ctaacctgat tgtggcgcag atgctgttcc tggaagcgga
      421 aaacccagaa aaagatatct atctgtacat taactcccca ggcggggtga tcactgccgg
      481 gatgtctatc tatgacacca tgcagtttat caagcctgat gtcagcacca tctgtatggg
      541 ccaggcggcc tcgatgggcg ctttcttgct gaccgcaggg gcaaaaggta aacgtttttg
      601 cctgccgaat tcgcgcgtga tgattcacca accgttgggc ggctaccagg gacaggcgac
      661 cgatatcgaa attcatgccc gtgaaattct gaaagttaaa gggcgcatga atgaacttat
      721 ggcgcttcat acgggtcaat cattagaaca gattgaacgt gataccgagc gcgatcgctt
      781 cctttccgcc cctgaagcgg tggaatacgg tctggtcgat tcgattctga cccatcgtaa
      841 ttgactcgag caaagcccgc cgaaaggcgg gcttttctgt gtcgaccgat gcccttgaga
      901 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
      961 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     1021 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1081 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1801 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1981 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     2581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2941 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3001 gccctttcgt cttcaagaat tctggcgaat cctctgacca gccagaaaac gacctttctg
     3061 tggtgaaacc ggatgctgca attcagagcg ccagcaagtg ggggacagca gaagacctga
     3121 ccgccgcaga gtggatgttt gacatggt
//
