LOCUS       sequence-197013-        3839 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 3839)
  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 3839)
  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..3839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(2513..2518,2519..2535,2536..2541))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2653..3198
                     /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     3099..3118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3315..3332
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3687..3706)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggcgaaaa cctggcccta
      181 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
      241 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
      301 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
      361 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
      421 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
      481 ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga aagcaaattc
      541 gacccgggcg gccgcaaccc agcgttcgat gcttctttga gcgaacgatc aaaaataagt
      601 gccttcccat caaaaaaata ttctcaacat aaaaaacttt gtgtaatact tgtaacgcta
      661 catggagatt aactcaatct agagggtaac tagtatggat aagaagccgc tggatgttct
      721 gatctctgcg accggtctgt ggatgtcccg taccggcacg ctgcacaaga tcaagcacta
      781 tgagatttct cgttctaaaa tctacatcga aatggcgtgt ggtgaccatc tggttgtgaa
      841 caactctcgt tcttgtcgtc ccgcacgtgc attccgttat cataaatacc gtaaaacctg
      901 caaacgttgt cgtgtttctg acgaagatat caacaacttc ctgacccgtt ctaccgaagg
      961 caaaacctct gttaaagtta aagttgtttc tgagccgaaa gtgaaaaaag cgatgccgaa
     1021 atctgtttct cgtgcgccga aaccgctgga aaatccggtt tctgcgaaag cgtctaccga
     1081 cacctctcgt tctgttccgt ctccggcgaa atctaccccg aactctccgg ttccgacctc
     1141 tgcgccggcg ccgtctctga cccgttctca gctggatcgt gttgaagcgc tgctgtctcc
     1201 ggaagataaa atctctctga acatcgcgaa accgttccgt gaactggaat ctgaactggt
     1261 tacccgtcgt aaaaacgatt tccagcgtct gtacaccaac gatcgtgaag actacctggg
     1321 taaactggaa cgtgacatca ccaaattctt cgttgaccgt gatttcctgg aaatcaaatc
     1381 tccgatcctg atcccggcgg aatacgttga acgtatgggt atcaacaacg ataccgaact
     1441 gtctaaacag atcttccgtg ttgataaaaa cctgtgcctg cgtccgatgc tggcgccgac
     1501 cctgtacaac tatctgcgta aactggatcg tatcctgccg gacccgatca aaatcttcga
     1561 agttggtccg tgctaccgta aagaatctga cggtaaagaa cacctggaag agttcaccat
     1621 ggtgaacttc tgccagatgg gttctggttg cacccgtgag aacctggaat ctctgatcaa
     1681 agaatttctg gactacctgg aaatcgactt cgaaatcgtt ggtgactcct gcatggtgta
     1741 cggtgatacc ctggacatca tgcacggtga cctggaactg tcttctgcgg ttgttggtcc
     1801 ggttccgctg gatcgtgaat ggggtatcga caaaccgtgg atcggtgcgg gtttcggtct
     1861 ggaacgtctg ctgaaagtta tgcacggttt caaaaacatc aaacgtgcgt ctcgttctga
     1921 atcttactac aacggtatct ctaccaacct gtaacctagg ctgctgccac cgctgagatc
     1981 tatgccccgc gcccacagga aggagctacc ggcagcggtg cggactgttg taactcataa
     2041 taagaaatga ggccgctcat ggcgttctgt tgcccgtctc gctggtgaaa agaaaaacca
     2101 ccctggctaa ctaagcggcc tgctgacttt ctcgccgatc aaaaggcatt ttgctattaa
     2161 gggattgacg agggcgtatc tgcgcagtaa gatgcgcccc gcattggaaa cctgatcatg
     2221 tagatcgaat ggactctaaa tccgttcagc cgggttagat tcccggggtt tccgccaaat
     2281 tcgaaaagcc tgctcaacga gcaggctttt ttgcatggcg ttacccaact taatcgcctt
     2341 gcagcacatc cccctttcgc cagatcccgc aagaggcccg gcagtaccgg cataaccaag
     2401 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     2461 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     2521 ttatcgatga taagctgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc
     2581 aggtgctaca tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat
     2641 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
     2701 aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg
     2761 taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc
     2821 gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt
     2881 ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2941 actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga
     3001 gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga
     3061 aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat
     3121 agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg
     3181 gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct
     3241 tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc
     3301 gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat
     3361 tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc
     3421 ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg
     3481 cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagta agaggttcca
     3541 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     3601 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     3661 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     3721 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     3781 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaactc
//
