LOCUS       sequence-348117-        3274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tRNA synthetase/tRNA pair for the in vivo incorporation of 
            7-Fluoro-L-Tryptophan (7FTrp) 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   Qianzhu H, Abdelkader EH, Herath ID, Otting G, Huber T
  TITLE     Site-Specific Incorporation of 7-Fluoro-L-tryptophan into Proteins 
            by Genetic Encoding to Monitor Ligand Binding by (19)F NMR 
            Spectroscopy.
  JOURNAL   ACS Sens. 2022 Jan 10. doi: 10.1021/acssensors.1c02467.
  PUBMED    35005899
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: "ACS Sens. 
            2022 Jan 10. doi: 10.1021/acssensors.1c02467."
COMMENT     Sequence Label: pRSF-G1(7FTrp)RS
FEATURES             Location/Qualifiers
     source          1..3274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(167..916)
                     /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(177..194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(417..436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(932..1747)
                     /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(932..1747)
                     /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     1655..1674
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1748..1839)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1922..1941
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1922..1940
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1972..1994
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2899..2917)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2913..2960
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cctttcgcca aattcgaaaa gcctgctcaa cgagcaggct tttttgcatg ctcgagcagc
       61 tcagggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
      121 cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaacttc cgcttcctcg
      181 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggtgtcagc tcactcaaaa
      241 gcggtaatac ggttatccac agaatcaggg gataaagccg gaaagaacat gtgagcaaaa
      301 agcaaagcac cggaagaagc caacgccgca ggcgtttttc cataggctcc gcccccctga
      361 cgagcatcac aaaaatcgac gctcaagcca gaggtggcga aacccgacag gactataaag
      421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      541 ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      661 aagacacgac ttatcgccac tggcagcagc cattggtaac tgatttagag gactttgtct
      721 tgaagttatg cacctgttaa ggctaaactg aaagaacaga ttttggtgag tgcggtcctc
      781 caacccactt accttggttc aaagagttgg tagctcagcg aaccttgaga aaaccaccgt
      841 tggtagcggt ggtttttctt tatttatgag atgatgaatc aatcggtcta tcaagtcaac
      901 gaacagctat tccgttgaac tgtggtacgc gttagaaaaa ctcatcgagc atcaaatgaa
      961 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1021 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     1081 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     1141 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     1201 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     1261 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc
     1321 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     1381 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     1441 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1501 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1561 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1621 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1681 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
     1741 ggctcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggcatgcag cgcttgactt
     1861 gacgggacgg cgttgtaatt ctcatgtttg acagcttatc atcgataagc ttggtaccca
     1921 ataatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
     1981 ctttaagaag gagatataca tatggtggtg aaatttaccg atagccagat tcagcatctg
     2041 atggaatatg gtgataatga ttggagcgaa gccgaatttg aagatgcagc agcacgtgat
     2101 aaagaattta gcagccagtt tagcaaactg aaaagcgcca atgataaagg cctgaaagat
     2161 gttattgcaa atccgcgtaa tgatctgacc gatctggaaa acaaaattcg cgaaaaactg
     2221 gcagcccgtg gttttattga agttcatacc ccgatttttg tgagcaaaag cgcactggca
     2281 aaaatgacca ttaccgaaga tcatccgctg ttcaaacagg tgttttggat tgatgataaa
     2341 cgtgcactgc gtccgatgca tgcaatgaat gggtttaaag ttatgcgtga actgcgcgat
     2401 cataccaaag gtccggttaa aatctttgaa attggtagct gctttcgcaa agaaagcaaa
     2461 agcagtaccc atctggaaga atttaccatg ctgggcctgt tcgaaatggg tcctgatggt
     2521 gatccgatgg aacatctgaa aatgtatatt ggcgatatca tggatgccgt tggtgttgaa
     2581 tataccacca gtcgtgaaga atcagatgtt tgggttgaaa ccctggacgt ggaaattaat
     2641 ggcaccgaag ttgcaagcgg tggggttggt ccgcataaac tggatccggc acatgatgtg
     2701 catgaaccgt atgcaggtat tggttttggt ctggaacgtc tgctgatgct gaaaaatggt
     2761 aaaagcaatg cacgcaaaac cggcaaaagt attacctatc tgaatggcta caaactggat
     2821 taagaattcg agctcccggg taccagatag atccggctgc taacaaagcc cgaaaggaag
     2881 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     2941 gggtcttgag gggttttttg gtttgtgagc tcccgtaatt ccgcttcgca acatgtgagc
     3001 accggtttat tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag
     3061 tttgctcagg ctctccccgt ggaggtaata attgacgata tgatcagtgc acggctaact
     3121 aagcggcctg ctgactttct cgccgatcaa aaggcatttt gctattaagg gattgacgag
     3181 ggcgtatctg cgcagtaaga tgcgccccgc attggagggc gctccggcga gcaaacgggt
     3241 ctctaaaacc tgtaagcggg gttcgacccc ccgg
//
