LOCUS       sequence-192359-        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: pDB038a
FEATURES             Location/Qualifiers
     source          1..6601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     757..774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    789..929
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(855..874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1015..1037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1031..1222)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             2871..3950
                     /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             4014..4994
                     /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(5692..6483)
                     /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(6484..6555)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggccaaatag gccgttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccacc tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      721 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      781 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      841 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
      901 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
      961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     1021 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     1081 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     1141 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     1201 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     1261 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacaaa
     1321 gaggacatcc ggtcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttgc
     1381 tgaggagact tagggaccct gatgaaattc gccacttgtt agtgtaattc gctaactcat
     1441 cctggcatgt tgctgttgat tcttcaatca gatctttata aatcaaaaag ataaaaaatt
     1501 ggcacgcaaa ttgtattaac agttcagcag gacaatcctg aacgcagaaa tcaagaggac
     1561 aacattacct gcaggtgcag taaggaggaa aaaaaaatga aaaaattatt attcgcaatt
     1621 cctttagttg ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa
     1681 tagcatacag aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac
     1741 gctaactatg agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa
     1801 actcagtgtt acggtacatg ggttcctatt gggcttgcta tccctgaaaa tgagggtggt
     1861 ggctctgagg gtggcggttc tgagggtggc ggttctgagg gtggcggtac taaacctcct
     1921 gagtacggtg atacacctat tccgggctat acttatatca accctctcga cggcacttat
     1981 ccgcctggta ctgagcaaaa ccccgctaat cctaatcctt ctcttgagga gtctcagcct
     2041 cttaatactt tcatgtttca gaataatagg ttccgaaata ggcagggggc attaactgtt
     2101 tatacgggca ctgttactca aggcactgac cccgttaaaa cttattagca gtacactcct
     2161 gtatcatcaa aagccatgta tgacgcttac tggaacggta aattcagaga ctgcgctttc
     2221 cattctggct ttaatgagga tccattcgtt tgtgaatatc aaggccaatc gtctgacctg
     2281 cctcaacctc ctgtcaatgc tggcggcggc tctggtggtg gttctggtgg cggctctgag
     2341 ggtggtggct ctgagggtgg cggttctgag ggtggcggct ctgagggagg cggttccggt
     2401 ggtggctctg gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct
     2461 atgaccgaaa atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct
     2521 gtcgctactg attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct
     2581 aatggtaatg gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt
     2641 gacggtgata attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa
     2701 tcggttgaat gtcgcccttt tgtctttggc gctggtaaac cttacgagtt cagtatcgac
     2761 tgcgataaga tcaacctgtt ccgcggtgtc tttgcgtttc ttttatatgt tgccaccttt
     2821 atgtatgtat tttctacgtt tgctaacata ctgcgtaata aggagtctta atgaaatttg
     2881 gaaacttttt gcttacatac caacctcccc aattttccca aacagaggta atgaaacgtt
     2941 tggttaaatt aggtcgcatc tctgaggagt gtggttttga taccgtatgg ttactggagc
     3001 atcatttcac ggagtttggt ttgcttggta acccttatgt cgctgctgca tatttacttg
     3061 gcgcgactaa aaaattgaat gtaggaactg ccgctattgt tcttcccaca gcccatccag
     3121 tacgccaact tgaagatgtg aatttattgg atcaaatgtc aaaaggacga tttcggtttg
     3181 gtatttgccg agggctttac aacaaggact ttcgcgtatt cggcacagat atgaataaca
     3241 gtcgcgcctt agcggaatgc tggtacgggc tgataaagaa tggcatgaca gagggatata
     3301 tggaagctga taatgaacat atcaagttcc ataaggtaaa agtaaacccc gcggcgtata
     3361 gcagaggtgg cgcaccggtt tatgtggtgg ctgaatcagc ttcgacgact gagtgggctg
     3421 ctcaatttgg cctaccgatg atattaagtt ggattataaa tactaacgaa aagaaagcac
     3481 aacttgagct ttataatgaa gtggctcaag aatatgggca cgatattcat aatatcgacc
     3541 attgcttatc atatataaca tctgtagatc atgactcaat taaagcgaaa gagatttgcc
     3601 ggaaatttct ggggcattgg tatgattctt atgtgaatgc tacgactatt tttgatgatt
     3661 cagaccaaac aagaggttat gatttcaata aagggcagtg gcgtgacttt gtattaaaag
     3721 gacataaaga tactaatcgc cgtattgatt acagttacga aatcaatccc gtgggaacgc
     3781 cgcaggaatg tattgacata attcaaaaag acattgatgc tacaggaata tcaaatattt
     3841 gttgtggatt tgaagctaat ggaacagtag acgaaattat tgcttccatg aagctcttcc
     3901 agtctgatgt catgccattt cttaaagaaa aacaacgttc gctattatat tatggcggtg
     3961 gcggtagcgg cggtggcggt agcggcggtg gcggtagcgg cggtggcggt agcaaatttg
     4021 gattgttctt ccttaacttc atcaattcaa caactgttca agaacagagt atagttcgca
     4081 tgcaggaaat aacggagtat gttgataagt tgaattttga acagatttta gtgtatgaaa
     4141 atcatttttc agataatggt gttgtcggcg ctcctctgac tgtttctggt tttctgctcg
     4201 gtttaacaga gaaaattaaa attggttcat taaatcacat cattacaact catcatcctg
     4261 tccgcatagc ggaggaagct tgcttattgg atcagttaag tgaagggaga tttattttag
     4321 ggtttagtga ttgcgaaaaa aaagatgaaa tgcatttttt taatcgcccg gttgaatatc
     4381 aacagcaact atttgaagag tgttatgaaa tcattaacga tgctttaaca acaggctatt
     4441 gtaatccaga taacgatttt tatagcttcc ctaaaatatc tgtaaatccc catgcttata
     4501 cgccaggcgg acctcggaaa tatgtaacag caaccagtca tcatattgtt gagtgggcgg
     4561 ccaaaaaagg tattcctctc atctttaagt gggatgattc taatgatgtt agatatgaat
     4621 atgctgaaag atataaagcc gttgcggata aatatgacgt tgacctatca gagatagacc
     4681 atcagttaat gatattagtt aactataacg aagatagtaa taaagctaaa caagagacgc
     4741 gtgcatttat tagtgattat gttcttgaaa tgcaccctaa tgaaaatttc gaaaataaac
     4801 ttgaagaaat aattgcagaa aacgctgtcg gaaattatac ggagtgtata actgcggcta
     4861 agttggcaat tgaaaagtgt ggtgcgaaaa gtgtattgct gtcctttgaa ccaatgaatg
     4921 atttgatgag ccaaaaaaat gtaatcaata ttgttgatga taatattaag aagtaccaca
     4981 cggaatatac ctaaacggag ccaatgtacg caaaaaaccc cgcttcagcg gggttttttc
     5041 gccaaaaaaa accccgccct gtcaggggcg gggttttttt ttagggccct actgactgtc
     5101 cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggtaataat
     5161 tgacgatatg atcagtgcac ggctaactaa gcggcctgct gactttctcg ccgatcaaaa
     5221 ggcattttgc tattaaggga ttgacgaggg cgtatctgcg cagtaagatg cgccccgcat
     5281 tcggaaacct gatcatgtag atcgaatgga ctctaaatcc gttcagccgg gttagattcc
     5341 cggggtttcc gccaggatcc gtcgacctgc agatccttag cgaaagctaa ggattttttt
     5401 taagcttctg ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta
     5461 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg
     5521 ttgtaattca ttaagcattc tgccgacgtt catcggatcc cgggcccgtc gactgcagag
     5581 gcctgcatgc aagcttggcg taatcatgat ggacttaatt aacggcactc ctcagcaaat
     5641 ataatgaccc tcttgataac ccaagagggc attttttaat gcccatggcg tttatttgcc
     5701 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     5761 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     5821 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     5881 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     5941 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     6001 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     6061 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     6121 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     6181 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     6241 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     6301 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     6361 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     6421 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     6481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtt tcatgagcgt
     6541 atacatattt gaatgtattt agaaaaatat taattgcgtt gcgctcactg cccgctttcc
     6601 a
//
