LOCUS       sequence-348119-        3274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tRNA synthetase/tRNA pair for the in vivo incorporation of 
            N6-(trifluoroacetyl)-L-lysine (TFA-Lys) 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 3274)
  AUTHORS   Orton HW, Qianzhu H, Abdelkader EH, Habel EI, Tan YJ, Frkic RL, 
            Jackson CJ, Huber T, Otting G
  TITLE     Through-Space Scalar (19)F-(19)F Couplings between Fluorinated 
            Noncanonical Amino Acids for the Detection of Specific Contacts in 
            Proteins.
  JOURNAL   J Am Chem Soc. 2021 Nov 15. doi: 10.1021/jacs.1c10104.
  PUBMED    34780162
REFERENCE   2  (bases 1 to 3274)
  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 Nov 15. doi: 10.1021/jacs.1c10104."
COMMENT     Sequence Label: pRSF-G1TFAKRS
FEATURES             Location/Qualifiers
     source          1..3274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(164..979)
                     /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(164..979)
                     /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     887..906
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(980..1071)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1154..1173
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1154..1172
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1204..1226
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2131..2149)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2145..2192
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(join(2673..3274,1..148))
                     /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(2683..2700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2923..2942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag cgaaccttga
       61 gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa tcaatcggtc
      121 tatcaagtca acgaacagct attccgttga actgtggtac gcgttagaaa aactcatcga
      181 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
      241 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
      301 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
      361 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
      421 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
      481 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
      541 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
      601 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
      661 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
      721 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
      781 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
      841 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
      901 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
      961 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1021 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
     1081 agcgcttgac ttgacgggac ggcgttgtaa ttctcatgtt tgacagctta tcatcgataa
     1141 gcttggtacc caataatacg actcactata gggagaccac aacggtttcc ctctagaaat
     1201 aattttgttt aactttaaga aggagatata catatggtgg tgaaatttac cgatagccag
     1261 attcagcatc tgatggaata tggtgataat gattggagcg aagccgaatt tgaagatgca
     1321 gcagcacgtg ataaagaatt tagcagccag tttagcaaac tgaaaagcgc caatgataaa
     1381 ggcctgaaag atgttattgc aaatccgcgt aatgatctga ccgatctgga aaacaaaatt
     1441 cgcgaaaaac tggcagcccg tggttttatt gaagttcata ccccgatttt tgtgagcaaa
     1501 agcgcactgg caaaaatgac cattaccgaa gatcatccgc tgttcaaaca ggtgttttgg
     1561 attgatgata aacgtgcact gcgtccgatg catgcaatga atattttgaa agttatgcgt
     1621 gaactgcgcg atcataccaa aggtccggtt aaaatctttg aaattggtag ctgctttcgc
     1681 aaagaaagca aaagcagtac ccatctggaa gaatttacca tgctgaacct ggccgaaatg
     1741 ggtcctgatg gtgatccgat ggaacatctg aaaatgtata ttggcgatat catggatgcc
     1801 gttggtgttg aatataccac cagtcgtgaa gaatcagatg tttgggttga aaccctggac
     1861 gtggaaatta atggcaccga agttgcaagc ggttgtgttg gtccgcataa actggatccg
     1921 gcacatgatg tgcatgaacc gtgggcaggt attggttttg gtctggaacg tctgctgatg
     1981 ctgaaaaatg gtaaaagcaa tgcacgcaaa accggcaaaa gtattaccta tctgaatggc
     2041 tacaaactgg attaagaatt cgagctcccg ggtaccagat agatccggct gctaacaaag
     2101 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     2161 gggcctctaa acgggtcttg aggggttttt tggtttgtga gctcccgtaa ttccgcttcg
     2221 caacatgtga gcaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat
     2281 gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga tatgatcagt
     2341 gcacggctaa ctaagcggcc tgctgacttt ctcgccgatc aaaaggcatt ttgctattaa
     2401 gggattgacg agggcgtatc tgcgcagtaa gatgcgcccc gcattggagg gcgctccggc
     2461 gagcaaacgg gtctctaaaa cctgtaagcg gggttcgacc ccccggcctt tcgccaaatt
     2521 cgaaaagcct gctcaacgag caggcttttt tgcatgctcg agcagctcag ggtcgaattt
     2581 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc
     2641 gtttaagggc accaataact gccttaaaaa aacttccgct tcctcgctca ctgactcgct
     2701 acgctcggtc gttcgactgc ggcgagcggt gtcagctcac tcaaaagcgg taatacggtt
     2761 atccacagaa tcaggggata aagccggaaa gaacatgtga gcaaaaagca aagcaccgga
     2821 agaagccaac gccgcaggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2881 atcgacgctc aagccagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt tggtatctca
     3061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3181 cgccactggc agcagccatt ggtaactgat ttagaggact ttgtcttgaa gttatgcacc
     3241 tgttaaggct aaactgaaag aacagatttt ggtg
//
