LOCUS       sequence-106824-        1028 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Beta-Lactamase protein in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1028)
  AUTHORS   D'Astolfo DS, Pagliero RJ, Pras A, Karthaus WR, Clevers H, Prasad V,
            Lebbink RJ, Rehmann H, Geijsen N
  TITLE     Efficient Intracellular Delivery of Native Proteins.
  JOURNAL   Cell. 2015 Apr 23;161(3):674-690. doi: 10.1016/j.cell.2015.03.028.
  PUBMED    25910214
REFERENCE   2  (bases 1 to 1028)
  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: 
            "10.1016/j.cell.2015.03"; journalName: "Cell"; date: "2015-04-23- 
            23"; volume: "161"; issue: "3"; pages: "674-690"
COMMENT     Sequence Label: Beta-Lactamase
FEATURES             Location/Qualifiers
     source          1..1028
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             20..42
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             61..78
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             88..105
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             130..924
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="MHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMM
                     STFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAIT
                     MSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMA
                     TTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGII
                     AALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     complement(282..301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(975..993)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat gggctcatcg
       61 catcatcatc atcatcattc atcgggtctg gttccgcgcg gttcacacgg tggtggttcg
      121 gcggccgcca tgcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
      181 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      241 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      301 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      361 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      421 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
      481 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
      541 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
      601 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
      661 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
      721 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
      781 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
      841 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
      901 gcctcactga ttaagcattg gtaaggatcc ggctgctaac aaagcccgaa aggaagctga
      961 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1021 cttgaggg
//
