LOCUS       sequence-327914-        4926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tRNA synthetase/tRNA pair for the in vivo incorporation of 
            para-Pentafluorosulfanyl-Phenylalanine, 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 4926)
  AUTHORS   Qianzhu H, Welegedara AP, Williamson H, McGrath AE, Mahawaththa MC, 
            Dixon NE, Otting G, Huber T
  TITLE     Genetic Encoding of para-Pentafluorosulfanyl Phenylalanine: A Highly
            Hydrophobic and Strongly Electronegative Group for Stable Protein 
            Interactions.
  JOURNAL   J Am Chem Soc. 2020 Oct 14;142(41):17277-17281. doi: 
            10.1021/jacs.0c07976. Epub 2020 Sep 30.
  PUBMED    32975937
REFERENCE   2  (bases 1 to 4926)
  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"; doi: 
            "10.1021/jacs.0c07976"; journalName: "J Am Chem Soc"; date: 
            "2020-10-14- 14"; volume: "142"; issue: "41"; pages: "17277-17281"
COMMENT     Sequence Label: pXPR-SF61
FEATURES             Location/Qualifiers
     source          1..4926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1249..1266
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1322..1339)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2043..2702)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2617..2636
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2703..2805)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3197..3214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3331..3876)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3411..3430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4022..4900)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctaacaggag gaattagatc tatggacgaa tttgaaatga taaagagaaa cacatctgaa
      361 attatcagcg aggaagagtt aagagaggtt ttaaaaaaag atgaaaaatc tgctgtgata
      421 ggttttgaac caagtggtaa aatacattta gggcattatc tccaaataaa aaagatgatt
      481 gatttacaaa atgctggatt tgatataatt atatcgttgg ctgatttaca tgcctattta
      541 aaccagaaag gagagttgga tgagattaga aaaataggag attataacaa aaaagttttt
      601 gaagcaatgg ggttaaaggc aaaatatgtt tatggaagtg aatgggcgct tgataaggat
      661 tatacactga atgtctatag attggcttta aaaactacct taaaaagagc aagaaggagt
      721 atggaactta tagcaagaga ggatgaaaat ccaaaggttg ctgaagttat ctatccaata
      781 atgcaggtta atggcgttca ttataagggc gttgatgttg cagttggggg gatggagcag
      841 agaaaaatac acatgttagc aagggagctt ttaccaaaaa aggttgtttg tattcacaac
      901 cctgtcttaa cgggtttgga tggagaagga aagatgagtt cttcaaaagg gaattttata
      961 gctgttgatg actctccaga agagattagg gctaagataa agaaagcata ctgcccagct
     1021 ggagttgttg aaggaaatcc aataatggag atagctaaat acttccttga atatccttta
     1081 accataaaaa ggccagaaaa atttggtgga gatttgacag ttaatagcta tgaggagtta
     1141 gagagtttat ttaaaaataa ggaattgcat ccaatgagat taaaaaatgc tgtagctgaa
     1201 gaacttataa agattttaga gccaattaga aagagattat aagtcgacca tcatcatcat
     1261 catcattgag tttaaacggt ctccagcttg gctgttttgg cggatgagag aagattttca
     1321 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     1381 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     1441 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     1501 cgaaaggctc agtcgaaaga ctgggccttg tttgtgagct cccggtcatc aatcatcccc
     1561 ataatccttg ttaattccgc ttcgcaacat gtgagcaccg gtttattgac taccggaagc
     1621 agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct ccccgtggag
     1681 gtaataattg acgatatgat cagtgcacgg ctaactaagc ggcctgctga ctttctcgcc
     1741 gatcaaaagg cattttgcta ttaagggatt gacgagggcg tatctgcgca gtaagatgcg
     1801 ccccgcattc cggcggtagt tcagcagggc agaacggcgg actctaaatc cgcatggcag
     1861 gggttcaaat cccctccgcc ggaccaaatt cgaaaagcct gctcaacgag caggcttttt
     1921 tgcatgctcg agcagctcag ggtcgaattt gctttcgaat ttctgccatt catccgctta
     1981 ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa
     2041 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     2101 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     2161 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     2221 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     2281 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     2341 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     2401 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     2461 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg
     2521 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     2581 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     2641 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     2701 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     2761 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc
     2821 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg
     2881 aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac
     2941 ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag
     3001 ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca
     3061 tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa
     3121 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca
     3181 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     3241 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     3301 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     3361 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     3481 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     3541 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     3601 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     3661 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     3721 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     3781 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     3841 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     3901 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     3961 atacgatata agttgtaatt ctcatgtttg acagcttatc atcgataagc ttggtaccca
     4021 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     4081 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     4141 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     4201 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     4261 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
     4321 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     4381 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
     4441 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     4501 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     4561 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     4621 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     4681 acagcaaaat atcactcggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     4741 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     4801 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     4861 agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat
     4921 tcagag
//
