LOCUS       sequence-194800-        3838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid encoding chPylRS(IPYE) and pylT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3838)
  AUTHORS   Bryson DI, Fan C, Guo LT, Miller C, Soll D, Liu DR
  TITLE     Continuous directed evolution of aminoacyl-tRNA synthetases.
  JOURNAL   Nat Chem Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474.
  PUBMED    29035361
REFERENCE   2  (bases 1 to 3838)
  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: "Nat Chem 
            Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474."
COMMENT     Sequence Label: pTECH-chPylRS(IPYE)
FEATURES             Location/Qualifiers
     source          1..3838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1958..1963,1964..1980,1981..1986))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2098..2643
                     /direction=RIGHT
                     /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     2544..2563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2760..2777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3066..3725
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3132..3151)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aacccagcgt tcgatgcttc tttgagcgaa cgatcaaaaa taagtgcctt cccatcaaaa
       61 aaatattctc aacataaaaa actttgtgta atacttgtaa cgctacatgg agattaactc
      121 aatctagagg gtaactagta tggataagaa gccgctggat gttctgatct ctgcgaccgg
      181 tctgtggatg tcccgtaccg gcacgctgca caagatcaag cactatgaga tttctcgttc
      241 taaaatctac atcgaaatgg cgtgtggtga ccatctggtt gtgaacaact ctcgttcttg
      301 tcgtcccgca cgtgcattcc gttatcataa ataccgtaaa acctgcaaac gttgtcgtgt
      361 ttctgacgaa gatatcaaca acttcctgac ccgttctacc gaaggcaaaa cctctgttaa
      421 agttaaagtt gtttctgagc cgaaagtgaa aaaagcgatg ccgaaatctg tttctcgtgc
      481 gccgaaaccg ctggaaaatc cggtttctgc gaaagcgtct accgacacct ctcgttctgt
      541 tccgtctccg gcgaaatcta ccccgaactc tccggttccg acctctgcaa gtgcccccgc
      601 acttacgaag agccagactg acaggcttga agtcctgtta aacccaaaag atgagatttc
      661 cctgaattcc ggcaagcctt tcagggagct tgagtccgaa ttgctctctc gcagaaaaaa
      721 agacctgcag cagatctacg cggaagaaag ggagaattat ctggggaaac tcgagcgtga
      781 aattaccagg ttctttgtgg acaggggttt tctggaaata aaatccccga tcctgatccc
      841 tcttgagtat atcgaaagga tgggcattga taatgatacc gaactttcaa aacagatctt
      901 cagggttgac aagaacttct gcctgagacc catgcttgct ccaaaccttt acaactacct
      961 gcgcaagctt gacagggccc tgcctgatcc aataaaaatt tttgaaatag gcccatgcta
     1021 cagaaaagag tccgacggca aagaacacct cgaagagttt accatgctga acttctgcca
     1081 gatgggatcg ggatgcacac gggaaaatct tgaaagcata attacggact tcctgaacca
     1141 cctgggaatt gatttcaaga tcgtaggcga ttcctgcatg gtctatgggg atacccttga
     1201 tgtaatgcac ggagacctgg aactttcctc tgcagtagtc ggacccatac cgcttgaccg
     1261 ggaatggggt attgataaac cctggatagg ggcaggtttc gggctcgaac gccttctaaa
     1321 ggttaaacac gactttaaaa atatcaagag agctgcaagg tccgagtctt actataacgg
     1381 gatttctacc aacctgtaac ctaggctgct gccaccgctg agatctatgg cccgcgccca
     1441 caggaaggag ctaccggcag cggtgcggac tgttgtaact cataataaga aatgaggccg
     1501 ctcatggcgt tctgttgccc gtctcgctgg tgaaaagaaa aaccaccctg gctaactaag
     1561 cggcctgctg actttctcgc cgatcaaaag gcattttgct attaagggat tgacgagggc
     1621 gtatctgcgc agtaagatgc gccccgcatt ggaaacctga tcatgtagat cgaatggact
     1681 ctaaatccgt tcagccgggt tagattcccg gggtttccgc caaattcgaa aagcctgctc
     1741 aacgagcagg cttttttgca tggcgttacc caacttaatc gccttgcagc acatccccct
     1801 ttcgccagat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
     1861 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
     1921 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
     1981 tgtcaaacat gagaattaca acttatatcg tatggggctg acttcaggtg ctacatttga
     2041 agagataaat tgcactgaaa tctagaaata ttttatctga ttaataagat gatcttcttg
     2101 agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg
     2161 cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg
     2221 agcgcagcca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac
     2281 taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg
     2341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt
     2401 cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa
     2461 tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga
     2521 gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2581 cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg
     2641 aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca
     2701 ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta
     2761 gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg
     2821 gtgcagcctt ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac
     2881 atagtaagcc agtatacact ccgctagcgc tgatgtccgg cggtgctttt gccgttacgc
     2941 accaccccgt cagtagctga acaggaggga cagtaagagg ttccaacttt caccataatg
     3001 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     3061 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     3121 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     3181 tggacattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     3241 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     3301 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     3361 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     3421 tatattcgca agatgtggcg tgttacggcg aaaacctggc ctatttccct aaagggttta
     3481 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     3541 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     3601 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     3661 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     3721 cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa
     3781 gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg gcggccgc
//
