LOCUS       sequence-327916-        3751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tRNA synthetase/tRNA pair for the in vivo incorporation of 
            N6-(((Trimethylsilyl)methoxy)carbonyl)-L-lysine (TMSK), into 
            proteins in E. coli in response to the amber (TAG) codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3751)
  AUTHORS   Abdelkader EH, Qianzhu H, Tan YJ, Adams LA, Huber T, Otting G
  TITLE     Genetic Encoding of 
            N(6)-(((Trimethylsilyl)methoxy)carbonyl)-l-lysine for NMR Studies of
            Protein-Protein and Protein-Ligand Interactions.
  JOURNAL   J Am Chem Soc. 2021 Jan 5. doi: 10.1021/jacs.0c11971.
  PUBMED    33399460
REFERENCE   2  (bases 1 to 3751)
  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: "J Am Chem 
            Soc. 2021 Jan 5. doi: 10.1021/jacs.0c11971."
COMMENT     Sequence Label: pRSF-ChPylTMSK
FEATURES             Location/Qualifiers
     source          1..3751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1347..1364
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1420..1437)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1441..1488
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1997..2746)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2007..2024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2247..2266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2762..3577)
                     /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             complement(2762..3577)
                     /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     3485..3504
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3578..3669)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccccc atcttagtat attagttaag
       61 tataagaagg agatatacat atggataaga agccgctgga tgttctgatc tctgcgaccg
      121 gtctgtggat gtcccgtacc ggcacgctgc acaagatcaa gcactatgag gtttctcgtt
      181 ctaaaatcta catcgaaatg gcgtgtggtg accatctggt tgtgaacaac tctcgttctt
      241 gtcgtaccgc acgtgcattc cgtcatcata aataccgtaa aacctgcaaa cgttgtcgtg
      301 tttctgacga agatatcaac aacttcctga cccgttctac cgaaggcaaa acctctgtta
      361 aagttaaagt tgtttctgcg ccgaaagtga aaaaagcgat gccgaaatct gtttctcgtg
      421 cgccgaaacc gctggaaaat ccggtttctg cgaaagcgtc taccgacacc tctcgttctg
      481 ttccgtctcc ggcgaaatct accccgaact ctccggttcc gacctctgca agtgcccccg
      541 cacttacgaa gagccagact gacaggcttg aagtcctgtt aaacccaaaa gatgagattt
      601 ccctgaattc cggcaagcct ttcagggagc ttgagtccga attgctctct cgcagaaaaa
      661 aagacctgca gcagatctac gcggaagaaa gggagaatta tctggggaaa ctcgagcgtg
      721 aaattaccag gttctttgtg gacaggggtt ttctggaaat aaaatccccg atcctgatcc
      781 ctcttgagta tatcgaaagg atgggcattg ataatgatac cgaactttca aaacagatct
      841 tcagggttga caagaacttc tgcctgagac ccatgcttgc tccaaacctt tacaactacc
      901 tgcgcaagct tgacagggcc ctgcctgatc caataaaaat ttttgaaata ggcccatgct
      961 acagaaaaga gtccgacggc aaagaacacc tcgaagagtt taccatgctg aatttcggtc
     1021 agatgggatc gggatgcaca cgggaaaatc ttgaaagcat aattacggac ttcctgaacc
     1081 acctgggaat tgatttcaag atcgtaggcg attcctgcat ggtctttggg gatacccttg
     1141 atgtaatgca cggagacctg gaactttcct ctgcatgtgt cggacccata ccgcttgacc
     1201 gggaatgggg tattgataaa ccctggatag gggcaggttt cggactcgaa cgccttctaa
     1261 aggttaaaca cgactttaaa aatatcaaga gagctgcaag gtccgagtct tactataacg
     1321 ggatttctac caacctgtaa gtcgaccatc atcatcatca tcattgagtt taaacggtct
     1381 ccagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     1441 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttggt ttgtgagctc
     1501 ccggtcatca atcatcccca taatccttgt taattccgct tcgcaacatg tgagcaccgg
     1561 tttattgact accggaagca gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc
     1621 tcaggctctc cccgtggagg taataattga cgatatgatc agtgcacggc taactaagcg
     1681 gcctgctgac tttctcgccg atcaaaaggc attttgctat taagggattg acgagggcgt
     1741 atctgcgcag taagatgcgc cccgcattgg aaacgtgatc atgtagatcg aacggactct
     1801 aaatccgttc agtggggtta gattccccac gtttccgcca aattcgaaaa gcctgctcaa
     1861 cgagcaggct tttttgcatg ctcgagcagc tcagggtcga atttgctttc gaatttctgc
     1921 cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat
     1981 aactgcctta aaaaaacttc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     2041 ctgcggcgag cggtgtcagc tcactcaaaa gcggtaatac ggttatccac agaatcaggg
     2101 gataaagccg gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc caacgccgca
     2161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagcca
     2221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2341 gggaagcgtg gcgctttctc atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt
     2401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2521 cattggtaac tgatttagag gactttgtct tgaagttatg cacctgttaa ggctaaactg
     2581 aaagaacaga ttttggtgag tgcggtcctc caacccactt accttggttc aaagagttgg
     2641 tagctcagcg aaccttgaga aaaccaccgt tggtagcggt ggtttttctt tatttatgag
     2701 atgatgaatc aatcggtcta tcaagtcaac gaacagctat tccgttgaac tgtggtacgc
     2761 gttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     2821 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     2881 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     2941 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     3001 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     3061 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     3121 gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg acaattacaa acaggaatcg
     3181 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     3241 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     3301 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     3361 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     3421 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     3481 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     3541 gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt
     3601 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3661 ataaacaaat aggcatgcag cgcttgactt gacgggacgg cgttgtaatt ctcatgtttg
     3721 acagcttatc atcgataagc ttggtaccca a
//
