LOCUS       sequence-317915-        4926 bp    DNA     circular SYN 14-APR-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 Apr 14, 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        2..286
                     /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     209..228
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1250..1267
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1323..1340)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2044..2703)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2618..2637
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2704..2806)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3198..3215)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3332..3877)
                     /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(3412..3431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4023..4901)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
       61 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
      121 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
      181 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
      241 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gtttttttgg
      301 gctaacagga ggaattagat ctatggacga atttgaaatg ataaagagaa acacatctga
      361 aattatcagc gaggaagagt taagagaggt tttaaaaaaa gatgaaaaat ctgctgtgat
      421 aggttttgaa ccaagtggta aaatacattt agggcattat ctccaaataa aaaagatgat
      481 tgatttacaa aatgctggat ttgatataat tatatcgttg gctgatttac atgcctattt
      541 aaaccagaaa ggagagttgg atgagattag aaaaatagga gattataaca aaaaagtttt
      601 tgaagcaatg gggttaaagg caaaatatgt ttatggaagt gaatgggcgc ttgataagga
      661 ttatacactg aatgtctata gattggcttt aaaaactacc ttaaaaagag caagaaggag
      721 tatggaactt atagcaagag aggatgaaaa tccaaaggtt gctgaagtta tctatccaat
      781 aatgcaggtt aatggcgttc attataaggg cgttgatgtt gcagttgggg ggatggagca
      841 gagaaaaata cacatgttag caagggagct tttaccaaaa aaggttgttt gtattcacaa
      901 ccctgtctta acgggtttgg atggagaagg aaagatgagt tcttcaaaag ggaattttat
      961 agctgttgat gactctccag aagagattag ggctaagata aagaaagcat actgcccagc
     1021 tggagttgtt gaaggaaatc caataatgga gatagctaaa tacttccttg aatatccttt
     1081 aaccataaaa aggccagaaa aatttggtgg agatttgaca gttaatagct atgaggagtt
     1141 agagagttta tttaaaaata aggaattgca tccaatgaga ttaaaaaatg ctgtagctga
     1201 agaacttata aagattttag agccaattag aaagagatta taagtcgacc atcatcatca
     1261 tcatcattga gtttaaacgg tctccagctt ggctgttttg gcggatgaga gaagattttc
     1321 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     1381 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     1441 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     1501 acgaaaggct cagtcgaaag actgggcctt gtttgtgagc tcccggtcat caatcatccc
     1561 cataatcctt gttaattccg cttcgcaaca tgtgagcacc ggtttattga ctaccggaag
     1621 cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga
     1681 ggtaataatt gacgatatga tcagtgcacg gctaactaag cggcctgctg actttctcgc
     1741 cgatcaaaag gcattttgct attaagggat tgacgagggc gtatctgcgc agtaagatgc
     1801 gccccgcatt ccggcggtag ttcagcaggg cagaacggcg gactctaaat ccgcatggca
     1861 ggggttcaaa tcccctccgc cggaccaaat tcgaaaagcc tgctcaacga gcaggctttt
     1921 ttgcatgctc gagcagctca gggtcgaatt tgctttcgaa tttctgccat tcatccgctt
     1981 attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa
     2041 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     2101 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     2161 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     2221 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     2281 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     2341 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     2401 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     2461 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     2521 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     2581 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     2641 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     2701 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     2761 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     2821 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     2881 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     2941 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     3001 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     3061 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     3121 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     3181 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     3241 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     3301 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     3361 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     3421 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     3481 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     3541 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     3601 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     3661 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     3721 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     3781 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     3841 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     3901 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     3961 catacgatat aagttgtaat tctcatgttt gacagcttat catcgataag cttggtaccc
     4021 aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg
     4081 ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa
     4141 ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc
     4201 tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa
     4261 agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa
     4321 ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc
     4381 ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga
     4441 tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca
     4501 aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca
     4561 aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac
     4621 tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg
     4681 aacagcaaaa tatcactcgg tcggcaaaca aattctcgtc cctgattttt caccaccccc
     4741 tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa
     4801 aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac
     4861 gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca
     4921 ttcaga
//
