LOCUS       sequence-192356-        6601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Accessory plasmid for PACE of PylRS variants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6601)
  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 6601)
  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: pDB038
FEATURES             Location/Qualifiers
     source          1..6601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    782..922
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(848..867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1008..1030
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1024..1215)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             2864..3943
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     CDS             4007..4987
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
                     SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
                     SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
                     PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
                     HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
                     AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
     CDS             complement(5685..6476)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(6477..6548)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taggccgttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc acctcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      841 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      901 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      961 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1021 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     1081 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     1141 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     1201 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     1261 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac aaagaggaca
     1321 tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tgctgaggag
     1381 acttagggac cctgatgaaa ttcgccactt gttagtgtaa ttcgctaact catcctggca
     1441 tgttgctgtt gattcttcaa tcagatcttt ataaatcaaa aagataaaaa attggcacgc
     1501 aaattgtatt aacagttcag caggacaatc ctgaacgcag aaatcaagag gacaacatta
     1561 cctgcaggtg cagtaaggag gaaaaaaaaa tgaaaaaatt attattcgca attcctttag
     1621 ttgttccttt ctattctcac tccgctgaaa ctgttgaaag ttgtttagca aaatagcata
     1681 cagaaaattc atttactaac gtctggaaag acgacaaaac tttagatcgt tacgctaact
     1741 atgagggctg tctgtggaat gctacaggcg ttgtagtttg tactggtgac gaaactcagt
     1801 gttacggtac atgggttcct attgggcttg ctatccctga aaatgagggt ggtggctctg
     1861 agggtggcgg ttctgagggt ggcggttctg agggtggcgg tactaaacct cctgagtacg
     1921 gtgatacacc tattccgggc tatacttata tcaaccctct cgacggcact tatccgcctg
     1981 gtactgagca aaaccccgct aatcctaatc cttctcttga ggagtctcag cctcttaata
     2041 ctttcatgtt tcagaataat aggttccgaa ataggcaggg ggcattaact gtttatacgg
     2101 gcactgttac tcaaggcact gaccccgtta aaacttatta ccagtacact cctgtatcat
     2161 caaaagccat gtatgacgct tactggaacg gtaaattcag agactgcgct ttccattctg
     2221 gctttaatga ggatccattc gtttgtgaat atcaaggcca atcgtctgac ctgcctcaac
     2281 ctcctgtcaa tgctggcggc ggctctggtg gtggttctgg tggcggctct gagggtggtg
     2341 gctctgaggg tggcggttct gagggtggcg gctctgaggg aggcggttcc ggtggtggct
     2401 ctggttccgg tgattttgat tatgaaaaga tggcaaacgc taataagggg gctatgaccg
     2461 aaaatgccga tgaaaacgcg ctacagtctg acgctaaagg caaacttgat tctgtcgcta
     2521 ctgattacgg tgctgctatc gatggtttca ttggtgacgt ttccggcctt gctaatggta
     2581 atggtgctac tggtgatttt gctggctcta attcccaaat ggctcaagtc ggtgacggtg
     2641 ataattcacc tttaatgaat aatttccgtc aatatttacc ttccctccct caatcggttg
     2701 aatgtcgccc ttttgtcttt ggcgctggta aaccttacga gttcagtatc gactgcgata
     2761 agatcaacct gttccgcggt gtctttgcgt ttcttttata tgttgccacc tttatgtatg
     2821 tattttctac gtttgctaac atactgcgta ataaggagtc ttaatgaaat ttggaaactt
     2881 tttgcttaca taccaacctc cccaattttc ccaaacagag gtaatgaaac gtttggttaa
     2941 attaggtcgc atctctgagg agtgtggttt tgataccgta tggttactgg agcatcattt
     3001 cacggagttt ggtttgcttg gtaaccctta tgtcgctgct gcatatttac ttggcgcgac
     3061 taaaaaattg aatgtaggaa ctgccgctat tgttcttccc acagcccatc cagtacgcca
     3121 acttgaagat gtgaatttat tggatcaaat gtcaaaagga cgatttcggt ttggtatttg
     3181 ccgagggctt tacaacaagg actttcgcgt attcggcaca gatatgaata acagtcgcgc
     3241 cttagcggaa tgctggtacg ggctgataaa gaatggcatg acagagggat atatggaagc
     3301 tgataatgaa catatcaagt tccataaggt aaaagtaaac cccgcggcgt atagcagagg
     3361 tggcgcaccg gtttatgtgg tggctgaatc agcttcgacg actgagtggg ctgctcaatt
     3421 tggcctaccg atgatattaa gttggattat aaatactaac gaaaagaaag cacaacttga
     3481 gctttataat gaagtggctc aagaatatgg gcacgatatt cataatatcg accattgctt
     3541 atcatatata acatctgtag atcatgactc aattaaagcg aaagagattt gccggaaatt
     3601 tctggggcat tggtatgatt cttatgtgaa tgctacgact atttttgatg attcagacca
     3661 aacaagaggt tatgatttca ataaagggca gtggcgtgac tttgtattaa aaggacataa
     3721 agatactaat cgccgtattg attacagtta cgaaatcaat cccgtgggaa cgccgcagga
     3781 atgtattgac ataattcaaa aagacattga tgctacagga atatcaaata tttgttgtgg
     3841 atttgaagct aatggaacag tagacgaaat tattgcttcc atgaagctct tccagtctga
     3901 tgtcatgcca tttcttaaag aaaaacaacg ttcgctatta tattatggcg gtggcggtag
     3961 cggcggtggc ggtagcggcg gtggcggtag cggcggtggc ggtagcaaat ttggattgtt
     4021 cttccttaac ttcatcaatt caacaactgt tcaagaacag agtatagttc gcatgcagga
     4081 aataacggag tatgttgata agttgaattt tgaacagatt ttagtgtatg aaaatcattt
     4141 ttcagataat ggtgttgtcg gcgctcctct gactgtttct ggttttctgc tcggtttaac
     4201 agagaaaatt aaaattggtt cattaaatca catcattaca actcatcatc ctgtccgcat
     4261 agcggaggaa gcttgcttat tggatcagtt aagtgaaggg agatttattt tagggtttag
     4321 tgattgcgaa aaaaaagatg aaatgcattt ttttaatcgc ccggttgaat atcaacagca
     4381 actatttgaa gagtgttatg aaatcattaa cgatgcttta acaacaggct attgtaatcc
     4441 agataacgat ttttatagct tccctaaaat atctgtaaat ccccatgctt atacgccagg
     4501 cggacctcgg aaatatgtaa cagcaaccag tcatcatatt gttgagtggg cggccaaaaa
     4561 aggtattcct ctcatcttta agtgggatga ttctaatgat gttagatatg aatatgctga
     4621 aagatataaa gccgttgcgg ataaatatga cgttgaccta tcagagatag accatcagtt
     4681 aatgatatta gttaactata acgaagatag taataaagct aaacaagaga cgcgtgcatt
     4741 tattagtgat tatgttcttg aaatgcaccc taatgaaaat ttcgaaaata aacttgaaga
     4801 aataattgca gaaaacgctg tcggaaatta tacggagtgt ataactgcgg ctaagttggc
     4861 aattgaaaag tgtggtgcga aaagtgtatt gctgtccttt gaaccaatga atgatttgat
     4921 gagccaaaaa aatgtaatca atattgttga tgataatatt aagaagtacc acacggaata
     4981 tacctaaacg gagccaatgt acgcaaaaaa ccccgcttca gcggggtttt ttcgccaaaa
     5041 aaaaccccgc cctgtcaggg gcggggtttt tttttagggc cctactgact gtccgtgtgc
     5101 ttctcaaatg cctgaggcca gtttgctcag gctctccccg tggaggtaat aattgacgat
     5161 atgatcagtg cacggctaac taagcggcct gctgactttc tcgccgatca aaaggcattt
     5221 tgctattaag ggattgacga gggcgtatct gcgcagtaag atgcgccccg cattcggaaa
     5281 cctgatcatg tagatcgaat ggactctaaa tccgttcagc cgggttagat tcccggggtt
     5341 tccgccagga tccgtcgacc tgcagatcct tagcgaaagc taaggatttt ttttaagctt
     5401 ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt ttaagggcac
     5461 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     5521 tcattaagca ttctgccgac gttcatcgga tcccgggccc gtcgactgca gaggcctgca
     5581 tgcaagcttg gcgtaatcat gatggactta attaacggca ctcctcagca aatataatga
     5641 ccctcttgat aacccaagag ggcatttttt aatgcccatg gcgtttattt gccgactacc
     5701 ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc
     5761 aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac
     5821 tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg
     5881 gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc
     5941 cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt
     6001 tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct
     6061 cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct
     6121 gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc
     6181 cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc
     6241 tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca
     6301 tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca
     6361 tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg
     6421 acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcatactc
     6481 ttcctttttc aatattattg aagcatttat cagggttatt gtttcatgag cgtatacata
     6541 tttgaatgta tttagaaaaa tattaattgc gttgcgctca ctgcccgctt tccaggccaa
     6601 a
//
