LOCUS       sequence-199822-        3838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid encoding MbPylRS(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-MbPylRS(IPYE)
FEATURES             Location/Qualifiers
     source          1..3838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1949..1954,1955..1971,1972..1977))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2089..2634
                     /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     2535..2554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2751..2768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3057..3716
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3123..3142)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttcgatgctt ctttgagcga acgatcaaaa ataagtgcct tcccatcaaa aaaatattct
       61 caacataaaa aactttgtgt aatacttgta acgctacatg gagattaact caatctagag
      121 ggtaactagt atggataaga agccgctgga tgttctgatc tctgcgaccg gtctgtggat
      181 gtcccgtacc ggcacgctgc acaagatcaa gcactatgag atttctcgtt ctaaaatcta
      241 catcgaaatg gcgtgtggtg accatctggt tgtgaacaac tctcgttctt gtcgtcccgc
      301 acgtgcattc cgttatcata aataccgtaa aacctgcaaa cgttgtcgtg tttctgacga
      361 agatatcaac aacttcctga cccgttctac cgaaggcaaa acctctgtta aagttaaagt
      421 tgtttctgag ccgaaagtga aaaaagcgat gccgaaatct gtttctcgtg cgccgaaacc
      481 gctggaaaat ccggtttctg cgaaagcgtc taccgacacc tctcgttctg ttccgtctcc
      541 ggcgaaatct accccgaact ctccggttcc gacctctgcg ccggcgccgt ctctgacccg
      601 ttctcagctg gatcgtgttg aagcgctgct gtctccggaa gataaaatct ctctgaacat
      661 cgcgaaaccg ttccgtgaac tggaatctga actggttacc cgtcgtaaaa acgatttcca
      721 gcgtctgtac accaacgatc gtgaagacta cctgggtaaa ctggaacgtg acatcaccaa
      781 attcttcgtt gaccgtgatt tcctggaaat caaatctccg atcctgatcc cggcggaata
      841 cgttgaacgt atgggtatca acaacgatac cgaactgtct aaacagatct tccgtgttga
      901 taaaaacctg tgcctgcgtc cgatgctggc gccgaccctg tacaactatc tgcgtaaact
      961 ggatcgtatc ctgccggacc cgatcaaaat cttcgaagtt ggtccgtgct accgtaaaga
     1021 atctgacggt aaagaacacc tggaagagtt caccatggtg aacttctgcc agatgggttc
     1081 tggttgcacc cgtgagaacc tggaatctct gatcaaagaa tttctggact acctggaaat
     1141 cgacttcgaa atcgttggtg actcctgcat ggtgtacggt gataccctgg acatcatgca
     1201 cggtgacctg gaactgtctt ctgcggttgt tggtccggtt ccgctggatc gtgaatgggg
     1261 tatcgacaaa ccgtggatcg gtgcgggttt cggtctggaa cgtctgctga aagttatgca
     1321 cggtttcaaa aacatcaaac gtgcgtctcg ttctgaatct tactacaacg gtatctctac
     1381 caacctgtaa cctaggctgc tgccaccgct gagatctatg ccccgcgccc acaggaagga
     1441 gctaccggca gcggtgcgga ctgttgtaac tcataataag aaatgaggcc gctcatggcg
     1501 ttctgttgcc cgtctcgctg gtgaaaagaa aaaccaccct ggctaactaa gcggcctgct
     1561 gactttctcg ccgatcaaaa ggcattttgc tattaaggga ttgacgaggg cgtatctgcg
     1621 cagtaagatg cgccccgcat tggaaacctg atcatgtaga tcgaatggac tctaaatccg
     1681 ttcagccggg ttagattccc ggggtttccg ccaaattcga aaagcctgct caacgagcag
     1741 gcttttttgc atggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccaga
     1801 tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg
     1861 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
     1921 tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca
     1981 tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg aagagataaa
     2041 ttgcactgaa atctagaaat attttatctg attaataaga tgatcttctt gagatcgttt
     2101 tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc
     2161 gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagcc
     2221 accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc
     2281 taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact
     2341 caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac
     2401 agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa
     2461 cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg
     2521 agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac
     2581 tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc
     2641 tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc
     2701 cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag
     2761 tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct
     2821 tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc
     2881 cagtatacac tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg caccaccccg
     2941 tcagtagctg aacaggaggg acagtaagag gttccaactt tcaccataat gaaataagat
     3001 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     3061 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     3121 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggacatta
     3181 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     3241 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     3301 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     3361 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     3421 aagatgtggc gtgttacggc gaaaacctgg cctatttccc taaagggttt attgagaata
     3481 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     3541 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     3601 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     3661 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt
     3721 tttaaggcag ttattggtgc ccttaaacgc ctggttgcta cgcctgaata agtgataata
     3781 agcggatgaa tggcagaaat tcgaaagcaa attcgacccg ggcggccgca acccagcg
//
