LOCUS       sequence-197049-        2746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NTerm 6xHis tagged protein L (PL) mutant K16TAG for 
            ncAA incoporation using the p70a promoter and UTR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2746)
  AUTHORS   Schlesinger O, Chemla Y, Heltberg M, Ozer E, Marshall R, Noireaux V,
            Jensen MH, Alfonta L
  TITLE     Tuning of Recombinant Protein Expression in Escherichia coli by 
            Manipulating Transcription, Translation Initiation Rates, and 
            Incorporation of Noncanonical Amino Acids.
  JOURNAL   ACS Synth Biol. 2017 Mar 9. doi: 10.1021/acssynbio.7b00019.
  PUBMED    28230975
REFERENCE   2  (bases 1 to 2746)
  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"; journalName: "ACS Synth 
            Biol. 2017 Mar 9. doi: 10.1021/acssynbio.7b00019."
COMMENT     Sequence Label: pBEST-p70a-UTR1-Nterm 6xHis PL K16TAG-T500
FEATURES             Location/Qualifiers
     source          1..2746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             260..282
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             295..312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(687..704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(858..1446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(938..957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1617..2408,2409..2477))
                     /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     2240..2259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2478..2582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2650..2668
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 attcagagcg ccagcaagtg ggggacagca gaagacctga ccgccgcaga gtggatgttt
       61 gacatggtga agactatcgc accatcagcc agaaaaccga attttgctgg gtgggctaac
      121 gatatccgcc tgatgcgtga acgtgacgga cgtaaccacc gcgacatgtg tgtgctgttc
      181 cgctgggcat gctgagctaa caccgtgcgt gttgacaatt ttacctctgg cggtgataat
      241 ggttgcagct agcaataatt ttgtttaact ttaagaagga gatataccat gggtcatcat
      301 catcatcatc atgccatgga agaagtaaca atctaggcta acctaatctt tgcaaatgga
      361 tccacacaaa ctgcagaatt caaaggaaca tttgaaaaag caactagtga agcttatgca
      421 tatgcagata ctttgaagaa agacaatgga gaatatactg tcgacgttgc agataaaggt
      481 tatactttaa atattaaatt tgctggatga ctcgagcaaa gcccgccgaa aggcgggctt
      541 ttctgtgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
      601 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
      661 aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      721 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      781 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      901 tgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1021 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1261 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1321 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1381 accgctggta acggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1441 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1501 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1561 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagtyac
     1621 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1681 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1741 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1801 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1861 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1921 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1981 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2041 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2101 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2161 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2221 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2281 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2341 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2401 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2461 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2521 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2581 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2641 acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattctg gcgaatcctc
     2701 tgaccagcca gaaaacgacc tttctgtggt gaaaccggat gctgca
//
