LOCUS       sequence-458352-        2986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mutant E. coli TrpRS from a weak glnS promoter for 
            proteome-wide incorporation of tryptophan analogues..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2986)
  AUTHORS   Loynd C, Singha Roy SJ, Canarelli SE, Bak DW, Sundaresh B, Babbitz 
            Z, van Opijnen T, Weerapana E, Chatterjee A
  TITLE     Cell-selective multiplexed bioorthogonal noncanonical amino acid 
            tagging for nascent proteomics.
  JOURNAL   Nat Chem Biol. 2025 Oct 2. doi: 10.1038/s41589-025-02039-3.
  PUBMED    41039161
REFERENCE   2  (bases 1 to 2986)
  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: "Nat Chem 
            Biol. 2025 Oct 2. doi: 10.1038/s41589-025-02039-3."
COMMENT     Sequence Label: pBK-EcTrpRS(H14)
FEATURES             Location/Qualifiers
     source          1..2986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1380..1968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1460..1479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2079..2098)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2090..2905
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2090..2905
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2163..2182)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cttttgctga gttgaaggat ccgcggccgc tcgggttgtc agcctgtccc gcttataaga
       61 tcatacgccg ttatacgttg tttacgcttt gaggaatccc atatgatgac taagcccatc
      121 gtttttgctg gcgcacagcc ctcaggtgaa ttgaccattg gtaactacat gggtgcgctg
      181 cgtcagtggg taaacatgca ggatgactac cattgcattt actgtatcgt tgaccaacac
      241 gcgatcaccg tgcgccagga tgcacagaag ctgcgtaaag cgacgctgga tacgctggcc
      301 ttgtatctgg cttgtggtat cgatcctgag aaaagcacca tttttgttca gtcccacgtg
      361 ccggaacatg cacagttagg ctgggcactg aactgctata cctacttcgg cgaactgagt
      421 cgcatgacgc agtttaaaga taaatctgcg cgttatgccg agaacatcaa cgctggtctg
      481 tttgactatc cggtgctgat ggcagcggac atcctgctgt atcaaactaa tctgggtcct
      541 tgtggtgaag accagaaaca gcacctcgaa ctgagccgcg atattgccca gcgtttcaac
      601 gcgctgtatg gcgagatctt taaggtgccg gagccgttta ttccgaaatc tggcgcgcgc
      661 gtaatgtcgc tgctggagcc gaccaagaag atgtccaagt ctgacgataa tcgcaataac
      721 gttatcggcc tgctggaaga tccgaaatcg gtagtgaaga aaatcaaacg tgcggtcact
      781 gactccgacg agccgccggt agttcgctac gatgtgcaga acaaagcggg cgtttccaac
      841 ctgttggata tcctttcagc ggtaacgggc cagagcatcc cagaactgga aaaacagttc
      901 gaaggcaaga tgtatggtca tctgaaaggt gaagtggctg atgccgtttc cggtatgctg
      961 actgaattgc aggaacgcta tcaccgtttc cgcaacgatg aagccttcct gcaacaggtg
     1021 atgaaagatg gcgcggaaaa agccagcgcg cacgcttccc gtacgctaaa agcggtgtac
     1081 gaagcgattg gttttgtggc gaagccgtaa ggtaccatgg ctgcagtttc aaacgctaaa
     1141 ttgcctgatg cgctacgctt atcaggccta catgatctct gcaatatatt gagtttgcgt
     1201 gcttttgtag gccggataag gcgttcacgc cgcatccggc aagaaacagc aaacaatcca
     1261 aaacgccgcg ttcagcggcg ttttttctgc ttttcttcgc gaattaattc cgcttcgcac
     1321 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1381 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1441 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1501 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1561 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1621 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1681 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1741 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1801 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1861 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1921 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1981 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2041 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2101 tcaacgggaa acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata
     2161 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     2221 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2281 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2341 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     2401 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2461 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2521 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2581 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2641 gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2701 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2761 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2821 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2881 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2941 cattacgctg acttgacggg acggcggctt tgttgaataa atcgaa
//
