LOCUS       sequence-182112-        3838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid encoding chPylRS(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-chPylRS(IPYE)
FEATURES             Location/Qualifiers
     source          1..3838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(2512..2517,2518..2534,2535..2540))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2652..3197
                     /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     3098..3117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3314..3331
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(3620..3838,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3686..3705)
                     /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 cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
      541 acccgggcgg ccgcaaccca gcgttcgatg cttctttgag cgaacgatca aaaataagtg
      601 ccttcccatc aaaaaaatat tctcaacata aaaaactttg tgtaatactt gtaacgctac
      661 atggagatta actcaatcta gagggtaact agtatggata agaagccgct ggatgttctg
      721 atctctgcga ccggtctgtg gatgtcccgt accggcacgc tgcacaagat caagcactat
      781 gagatttctc gttctaaaat ctacatcgaa atggcgtgtg gtgaccatct ggttgtgaac
      841 aactctcgtt cttgtcgtcc cgcacgtgca ttccgttatc ataaataccg taaaacctgc
      901 aaacgttgtc gtgtttctga cgaagatatc aacaacttcc tgacccgttc taccgaaggc
      961 aaaacctctg ttaaagttaa agttgtttct gagccgaaag tgaaaaaagc gatgccgaaa
     1021 tctgtttctc gtgcgccgaa accgctggaa aatccggttt ctgcgaaagc gtctaccgac
     1081 acctctcgtt ctgttccgtc tccggcgaaa tctaccccga actctccggt tccgacctct
     1141 gcaagtgccc ccgcacttac gaagagccag actgacaggc ttgaagtcct gttaaaccca
     1201 aaagatgaga tttccctgaa ttccggcaag cctttcaggg agcttgagtc cgaattgctc
     1261 tctcgcagaa aaaaagacct gcagcagatc tacgcggaag aaagggagaa ttatctgggg
     1321 aaactcgagc gtgaaattac caggttcttt gtggacaggg gttttctgga aataaaatcc
     1381 ccgatcctga tccctcttga gtatatcgaa aggatgggca ttgataatga taccgaactt
     1441 tcaaaacaga tcttcagggt tgacaagaac ttctgcctga gacccatgct tgctccaaac
     1501 ctttacaact acctgcgcaa gcttgacagg gccctgcctg atccaataaa aatttttgaa
     1561 ataggcccat gctacagaaa agagtccgac ggcaaagaac acctcgaaga gtttaccatg
     1621 ctgaacttct gccagatggg atcgggatgc acacgggaaa atcttgaaag cataattacg
     1681 gacttcctga accacctggg aattgatttc aagatcgtag gcgattcctg catggtctat
     1741 ggggataccc ttgatgtaat gcacggagac ctggaacttt cctctgcagt agtcggaccc
     1801 ataccgcttg accgggaatg gggtattgat aaaccctgga taggggcagg tttcgggctc
     1861 gaacgccttc taaaggttaa acacgacttt aaaaatatca agagagctgc aaggtccgag
     1921 tcttactata acgggatttc taccaacctg taacctaggc tgctgccacc gctgagatct
     1981 atggcccgcg cccacaggaa ggagctaccg gcagcggtgc ggactgttgt aactcataat
     2041 aagaaatgag gccgctcatg gcgttctgtt gcccgtctcg ctggtgaaaa gaaaaaccac
     2101 cctggctaac taagcggcct gctgactttc tcgccgatca aaaggcattt tgctattaag
     2161 ggattgacga gggcgtatct gcgcagtaag atgcgccccg cattggaaac ctgatcatgt
     2221 agatcgaatg gactctaaat ccgttcagcc gggttagatt cccggggttt ccgccaaatt
     2281 cgaaaagcct gctcaacgag caggcttttt tgcatggcgt tacccaactt aatcgccttg
     2341 cagcacatcc ccctttcgcc agatcccgca agaggcccgg cagtaccggc ataaccaagc
     2401 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
     2461 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     2521 tatcgatgat aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca
     2581 ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata
     2641 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     2701 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     2761 aactggcttg gaggagcgca gccaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     2821 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     2881 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     2941 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     3001 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     3061 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     3121 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     3181 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     3241 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     3301 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     3361 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     3421 tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc
     3481 ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga gggacagtaa gaggttccaa
     3541 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     3601 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     3661 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     3721 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     3781 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaactc
//
