LOCUS       sequence-199821-        3943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid encoding MmAcK3RS(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-MmAcK3RS(IPYE)
FEATURES             Location/Qualifiers
     source          1..3943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(2062..2067,2068..2084,2085..2090))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2202..2747
                     /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     2648..2667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2864..2881
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3170..3829
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3236..3255)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 acccagcgtt cgatgcttct ttgagcgaac gatcaaaaat aagtgccttc ccatcaaaaa
       61 aatattctca acataaaaaa ctttgtgtaa tacttgtaac gctacatgga gattaactca
      121 atctagaggg taactagtat ggataaaaaa ccactaaaca ctctgatatc tgcaaccggg
      181 ctctggatgt ccaggaccgg aacaattcat aaaataaaac accacgaaat ctctcgaagc
      241 aaaatctata ttgaaatggc atgcggagac caccttgttg taaacaactc caggagcagc
      301 aggcctgcaa gagcgctcag gtaccacaaa tacaggaaga cctgcaaacg ctgcagggtt
      361 tcgggtgagg atctcaataa gttcctcaca aaggcaaacg aagaccagac aagcgtaaaa
      421 gtcaaggtcg tttctgagcc taccagaacg aaaaaggcaa tgccaaaatc cgttgcgaga
      481 gccccgaaac ctcttgagaa tacagaagcg gcacaggctc aaccttctgg atctaaattt
      541 tcacctgcga taccggtttc cacccaagag tcagtttctg tcccggcatc tgtttcaaca
      601 tcaatatcaa gcatttctac aggagcaact gcatccgcac tggtaaaagg gaatacgaat
      661 cccattacat ccatgtctgc ccctgttcag gcaagtgccc ccgcacttac gaagagccag
      721 actgacaggc ttgaagtcct gttaaaccca aaagatgaga tttccctgaa ttccggcaag
      781 cctttcaggg agcttgagtc cgaattgctc tctcgcagaa aaaaagacct gcagcagatc
      841 tacgcggaag aaagggagaa ttatctgggg aaactcgagc gtgaaattac caggttcttt
      901 gtggacaggg gttttctgga aataaaatcc ccgatcctga tccctcttga gtatatcgaa
      961 aggatgggca ttgataatga taccgaactt tcaaaacaga tcttcagggt tgacaagaac
     1021 ttctgcctga gacccatgat ggctccaaac atttttaact acgctcgcaa gcttgacagg
     1081 gccctgcctg atccaataaa aatttttgaa ataggcccat gctacagaaa agagtccgac
     1141 ggcaaagaac acctcgaaga gtttaccatg ctgaacttct ttcagatggg atcgggatgc
     1201 acacgggaaa atcttgaaag cataattacg gacttcctga accacctggg aattgatttc
     1261 aagatcgtag gcgattcctg catggtctat ggggataccc ttgatgtaat gcacggagac
     1321 ctggaacttt cctctgcagt agtcggaccc ataccgcttg accgggaatg gggtattgat
     1381 aaaccctgga taggggcagg tttcgggctc gaacgccttc taaaggttaa acacgacttt
     1441 aaaaatatca agagagctgc aaggtccgag tcttactata acgggatttc taccaacctg
     1501 taacctaggc tgctgccacc gctgagatct atgccccgcg cccacaggaa ggagctaccg
     1561 gcagcggtgc ggactgttgt aactcataat aagaaatgag gccgctcatg gcgttctgtt
     1621 gcccgtctcg ctggtgaaaa gaaaaaccac cctggctaac taagcggcct gctgactttc
     1681 tcgccgatca aaaggcattt tgctattaag ggattgacga gggcgtatct gcgcagtaag
     1741 atgcgccccg cattggaaac ctgatcatgt agatcgaatg gactctaaat ccgttcagcc
     1801 gggttagatt cccggggttt ccgccaaatt cgaaaagcct gctcaacgag caggcttttt
     1861 tgcatggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agatcccgca
     1921 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
     1981 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
     2041 ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat
     2101 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
     2161 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
     2221 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     2281 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gccaccaaaa
     2341 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     2401 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     2461 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     2521 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     2581 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     2641 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     2701 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     2761 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     2821 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     2881 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     2941 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     3001 cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag
     3061 ctgaacagga gggacagtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     3121 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     3181 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     3241 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggaca ttacggcctt
     3301 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     3361 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     3421 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     3481 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     3541 ggcgtgttac ggcgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     3601 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     3661 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     3721 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     3781 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg
     3841 cagttattgg tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat
     3901 gaatggcaga aattcgaaag caaattcgac ccgggcggcc gca
//
