LOCUS       sequence-199828-        3943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid encoding MmPylRS(IPYE) and pylT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3943)
  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 3943)
  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-MmPylRS(IPYE)
FEATURES             Location/Qualifiers
     source          1..3943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..546
                     /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     447..466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     663..680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             969..1628
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1035..1054)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3804..3809,3810..3826,3827..3832))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
       61 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
      121 aggagcgcag ccaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
      181 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
      241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
      301 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
      361 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
      421 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
      481 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
      541 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
      601 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
      661 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
      721 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
      781 aacatagtaa gccagtatac actccgctag cgctgatgtc cggcggtgct tttgccgtta
      841 cgcaccaccc cgtcagtagc tgaacaggag ggacagtaag aggttccaac tttcaccata
      901 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
      961 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     1021 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     1081 agctggacat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     1141 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga
     1201 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     1261 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     1321 acatatattc gcaagatgtg gcgtgttacg gcgaaaacct ggcctatttc cctaaagggt
     1381 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     1441 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     1501 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     1561 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     1621 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
     1681 taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc cgggcggccg
     1741 caacccagcg ttcgatgctt ctttgagcga acgatcaaaa ataagtgcct tcccatcaaa
     1801 aaaatattct caacataaaa aactttgtgt aatacttgta acgctacatg gagattaact
     1861 caatctagag ggtaactagt atggataaaa aaccactaaa cactctgata tctgcaaccg
     1921 ggctctggat gtccaggacc ggaacaattc ataaaataaa acaccacgaa atctctcgaa
     1981 gcaaaatcta tattgaaatg gcatgcggag accaccttgt tgtaaacaac tccaggagca
     2041 gcaggcctgc aagagcgctc aggtaccaca aatacaggaa gacctgcaaa cgctgcaggg
     2101 tttcggatga ggatctcaat aagttcctca caaaggcaaa cgaagaccag acaagcgtaa
     2161 aagtcaaggt cgtttctgaa cctaccagaa cgaaaaaggc aatgccaaaa tccgttgcga
     2221 gagccccgaa acctcttgag aatacagaag cggcacaggc tcaaccttct ggatctaaat
     2281 tttcacctgc gataccggtt tccacccaag agtcagtttc tgtcccggca tctgtttcaa
     2341 catcaatatc aagcatttct acaggagcaa ctgcatccgc actggtaaaa gggaatacga
     2401 atcccattac atccatgtct gcccctgttc aggcaagtgc ccccgcactt acgaagagcc
     2461 agactgacag gcttgaagtc ctgttaaacc caaaagatga gatttccctg aattccggca
     2521 agcctttcag ggagcttgag tccgaattgc tctctcgcag aaaaaaagac ctgcagcaga
     2581 tctacgcgga agaaagggag aattatctgg ggaaactcga gcgtgaaatt accaggttct
     2641 ttgtggacag gggttttctg gaaataaaat ccccgatcct gatccctctt gagtatatcg
     2701 aaaggatggg cattgataat gataccgaac tttcaaaaca gatcttcagg gttgacaaga
     2761 acttctgcct gagacccatg cttgctccaa acctttacaa ctacctgcgc aagcttgaca
     2821 gggccctgcc tgatccaata aaaatttttg aaataggccc atgctacaga aaagagtccg
     2881 acggcaaaga acacctcgaa gagtttacca tgctgaactt ctgccagatg ggatcgggat
     2941 gcacacggga aaatcttgaa agcataatta cggacttcct gaaccacctg ggaattgatt
     3001 tcaagatcgt aggcgattcc tgcatggtct atggggatac ccttgatgta atgcacggag
     3061 acctggaact ttcctctgca gtagtcggac ccataccgct tgaccgggaa tggggtattg
     3121 ataaaccctg gataggggca ggtttcgggc tcgaacgcct tctaaaggtt aaacacgact
     3181 ttaaaaatat caagagagct gcaaggtccg agtcttacta taacgggatt tctaccaacc
     3241 tgtaacctag gctgctgcca ccgctgagat ctatgccccg cgcccacagg aaggagctac
     3301 cggcagcggt gcggactgtt gtaactcata ataagaaatg aggccgctca tggcgttctg
     3361 ttgcccgtct cgctggtgaa aagaaaaacc accctggcta actaagcggc ctgctgactt
     3421 tctcgccgat caaaaggcat tttgctatta agggattgac gagggcgtat ctgcgcagta
     3481 agatgcgccc cgcattggaa acctgatcat gtagatcgaa tggactctaa atccgttcag
     3541 ccgggttaga ttcccggggt ttccgccaaa ttcgaaaagc ctgctcaacg agcaggcttt
     3601 tttgcatggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagatcccg
     3661 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     3721 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     3781 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     3841 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     3901 ctgaaatcta gaaatatttt atctgattaa taagatgatc ttc
//
