LOCUS       sequence-199816-        3838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid encoding MbAcK3RS(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-MbAcK3RS(IPYE)
FEATURES             Location/Qualifiers
     source          1..3838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1899..1904,1905..1921,1922..1927))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2039..2584
                     /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     2485..2504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2701..2718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3007..3666
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3073..3092)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aaaatattct caacataaaa aactttgtgt aatacttgta acgctacatg gagattaact
       61 caatctagag ggtaactagt atggataaga agccgctgga tgttctgatc tctgcgaccg
      121 gtctgtggat gtcccgtacc ggcacgctgc acaagatcaa gcactatgag atttctcgtt
      181 ctaaaatcta catcgaaatg gcgtgtggtg accatctggt tgtgaacaac tctcgttctt
      241 gtcgtcccgc acgtgcattc cgttatcata aataccgtaa aacctgcaaa cgttgtcgtg
      301 tttctggtga agatatcaac aacttcctga cccgttctac cgaaggcaaa acctctgtta
      361 aagttaaagt tgtttctgag ccgaaagtga aaaaagcgat gccgaaatct gtttctcgtg
      421 cgccgaaacc gctggaaaat ccggtttctg cgaaagcgtc taccgacacc tctcgttctg
      481 ttccgtctcc ggcgaaatct accccgaact ctccggttcc gacctctgcg ccggcgccgt
      541 ctctgacccg ttctcagctg gatcgtgttg aagcgctgct gtctccggaa gataaaatct
      601 ctctgaacat cgcgaaaccg ttccgtgaac tggaatctga actggttacc cgtcgtaaaa
      661 acgatttcca gcgtctgtac accaacgatc gtgaagacta cctgggtaaa ctggaacgtg
      721 acatcaccaa attcttcgtt gaccgtgatt tcctggaaat caaatctccg atcctgatcc
      781 cggcggaata cgttgaacgt atgggtatca acaacgatac cgaactgtct aaacagatct
      841 tccgtgttga taaaaacctg tgcctgcgtc cgatgatggc gccgaccatt tttaactatg
      901 ctcgtaaact ggatcgtatc ctgccggacc cgatcaaaat cttcgaagtt ggtccgtgct
      961 accgtaaaga atctgacggt aaagaacacc tggaagagtt caccatggtg aacttctttc
     1021 agatgggttc tggttgcacc cgtgagaacc tggaatctct gatcaaagaa tttctggact
     1081 acctggaaat cgacttcgaa atcgttggtg actcctgcat ggtgtacggt gataccctgg
     1141 acatcatgca cggtgacctg gaactgtctt ctgcggttgt tggtccggtt ccgctggatc
     1201 gtgaatgggg tatcgacaaa ccgtggatcg gtgcgggttt cggtctggaa cgtctgctga
     1261 aagttatgca cggtttcaaa aacatcaaac gtgcgtctcg ttctgaatct tactacaacg
     1321 gtatctctac caacctgtaa cctaggctgc tgccaccgct gagatctatg ccccgcgccc
     1381 acaggaagga gctaccggca gcggtgcgga ctgttgtaac tcataataag aaatgaggcc
     1441 gctcatggcg ttctgttgcc cgtctcgctg gtgaaaagaa aaaccaccct ggctaactaa
     1501 gcggcctgct gactttctcg ccgatcaaaa ggcattttgc tattaaggga ttgacgaggg
     1561 cgtatctgcg cagtaagatg cgccccgcat tggaaacctg atcatgtaga tcgaatggac
     1621 tctaaatccg ttcagccggg ttagattccc ggggtttccg ccaaattcga aaagcctgct
     1681 caacgagcag gcttttttgc atggcgttac ccaacttaat cgccttgcag cacatccccc
     1741 tttcgccaga tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc
     1801 atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc
     1861 ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag
     1921 ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg
     1981 aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga tgatcttctt
     2041 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
     2101 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     2161 gagcgcagcc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     2221 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     2281 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     2341 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     2401 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     2461 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2521 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     2581 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     2641 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     2701 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     2761 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     2821 catagtaagc cagtatacac tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg
     2881 caccaccccg tcagtagctg aacaggaggg acagtaagag gttccaactt tcaccataat
     2941 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     3001 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     3061 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     3121 ctggacatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     3181 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     3241 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     3301 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     3361 atatattcgc aagatgtggc gtgttacggc gaaaacctgg cctatttccc taaagggttt
     3421 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     3481 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     3541 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     3601 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     3661 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggttgcta cgcctgaata
     3721 agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg ggcggccgca
     3781 acccagcgtt cgatgcttct ttgagcgaac gatcaaaaat aagtgccttc ccatcaaa
//
