LOCUS       sequence-198819-        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Accessory plasmid for PACE of MjTyrRS variants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7061)
  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 7061)
  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: pDB007(+)
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..85
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        66..84
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             122..1396
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     CDS             1396..2475
                     /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             2539..3519
                     /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(join(4271..5062,5063..5131))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4894..4913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5132..5236)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5304..5322
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5343..6293)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      6341..6563
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 taaaacgaaa ggctcagtcg aaagacttgg cctttcgttt tgctgaggag acttagggac
       61 cctactaata cgactcacta tagggagaaa gacctgcagg tgcagtaagg aggaaaaaaa
      121 aatgaaaaaa ttattattcg caattccttt agttgttcct ttctattctc actccgctga
      181 aactgttgaa agttgtttag caaaacccca tacagaaaat tcatttacta acgtctggaa
      241 agacgacaaa actttagatc gttacgctaa ctatgagggc tgtctgtgga atgctacagg
      301 cgttgtagtt tgtactggtg acgaaactca gtgttacggt acatgggttc ctattgggct
      361 tgctatccct gaaaatgagg gtggtggctc tgagggtggc ggttctgagg gtggcggttc
      421 tgagggtggc ggtactaaac ctcctgagta cggtgataca cctattccgg gctatactta
      481 tatcaaccct ctcgacggca cttatccgcc tggtactgag caaaaccccg ctaatcctaa
      541 tccttctctt gaggagtctc agcctcttaa tactttcatg tttcagaata ataggttccg
      601 aaataggcag ggggcattaa ctgtttatac gggcactgtt actcaaggca ctgaccccgt
      661 taaaacttat taccagtaca ctcctgtatc atcaaaagcc atgtatgacg cttactggaa
      721 cggtaaattc agagactgcg ctttccattc tggctttaat gaggatccat tcgtttgtga
      781 atatcaaggc caatcgtctg acctgcctca acctcctgtc aatgctggcg gcggctctgg
      841 tggtggttct ggtggcggct ctgagggtgg tggctctgag ggtggcggtt ctgagggtgg
      901 cggctctgag ggaggcggtt ccggtggtgg ctctggttcc ggtgattttg attatgaaaa
      961 gatggcaaac gctaataagg gggctatgac cgaaaatgcc gatgaaaacg cgctacagtc
     1021 tgacgctaaa ggcaaacttg attctgtcgc tactgattac ggtgctgcta tcgatggttt
     1081 cattggtgac gtttccggcc ttgctaatgg taatggtgct actggtgatt ttgctggctc
     1141 taattcccaa atggctcaag tcggtgacgg tgataattca cctttaatga ataatttccg
     1201 tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc ccttttgtct ttggcgctgg
     1261 taaaccttac gagttcagta tcgactgcga taagatcaac ctgttccgcg gtgtctttgc
     1321 gtttctttta tatgttgcca cctttatgta tgtattttct acgtttgcta acatactgcg
     1381 taataaggag tcttaatgaa atttggaaac tttttgctta cataccaacc tccccaattt
     1441 tcccaaacag aggtaatgaa acgtttggtt aaattaggtc gcatctctga ggagtgtggt
     1501 tttgataccg tatggttact ggagcatcat ttcacggagt ttggtttgct tggtaaccct
     1561 tatgtcgctg ctgcatattt acttggcgcg actaaaaaat tgaatgtagg aactgccgct
     1621 attgttcttc ccacagccca tccagtacgc caacttgaag atgtgaattt attggatcaa
     1681 atgtcaaaag gacgatttcg gtttggtatt tgccgagggc tttacaacaa ggactttcgc
     1741 gtattcggca cagatatgaa taacagtcgc gccttagcgg aatgctggta cgggctgata
     1801 aagaatggca tgacagaggg atatatggaa gctgataatg aacatatcaa gttccataag
     1861 gtaaaagtaa accccgcggc gtatagcaga ggtggcgcac cggtttatgt ggtggctgaa
     1921 tcagcttcga cgactgagtg ggctgctcaa tttggcctac cgatgatatt aagttggatt
     1981 ataaatacta acgaaaagaa agcacaactt gagctttata atgaagtggc tcaagaatat
     2041 gggcacgata ttcataatat cgaccattgc ttatcatata taacatctgt agatcatgac
     2101 tcaattaaag cgaaagagat ttgccggaaa tttctggggc attggtatga ttcttatgtg
     2161 aatgctacga ctatttttga tgattcagac caaacaagag gttatgattt caataaaggg
     2221 cagtggcgtg actttgtatt aaaaggacat aaagatacta atcgccgtat tgattacagt
     2281 tacgaaatca atcccgtggg aacgccgcag gaatgtattg acataattca aaaagacatt
     2341 gatgctacag gaatatcaaa tatttgttgt ggatttgaag ctaatggaac agtagacgaa
     2401 attattgctt ccatgaagct cttccagtct gatgtcatgc catttcttaa agaaaaacaa
     2461 cgttcgctat tatattatgg cggtggcggt agcggcggtg gcggtagcgg cggtggcggt
     2521 agcggcggtg gcggtagcaa atttggattg ttcttcctta acttcatcaa ttcaacaact
     2581 gttcaagaac agagtatagt tcgcatgcag gaaataacgg agtatgttga taagttgaat
     2641 tttgaacaga ttttagtgta tgaaaatcat ttttcagata atggtgttgt cggcgctcct
     2701 ctgactgttt ctggttttct gctcggttta acagagaaaa ttaaaattgg ttcattaaat
     2761 cacatcatta caactcatca tcctgtccgc atagcggagg aagcttgctt attggatcag
     2821 ttaagtgaag ggagatttat tttagggttt agtgattgcg aaaaaaaaga tgaaatgcat
     2881 ttttttaatc gcccggttga atatcaacag caactatttg aagagtgtta tgaaatcatt
     2941 aacgatgctt taacaacagg ctattgtaat ccagataacg atttttatag cttccctaaa
     3001 atatctgtaa atccccatgc ttatacgcca ggcggacctc ggaaatatgt aacagcaacc
     3061 agtcatcata ttgttgagtg ggcggccaaa aaaggtattc ctctcatctt taagtgggat
     3121 gattctaatg atgttagata tgaatatgct gaaagatata aagccgttgc ggataaatat
     3181 gacgttgacc tatcagagat agaccatcag ttaatgatat tagttaacta taacgaagat
     3241 agtaataaag ctaaacaaga gacgcgtgca tttattagtg attatgttct tgaaatgcac
     3301 cctaatgaaa atttcgaaaa taaacttgaa gaaataattg cagaaaacgc tgtcggaaat
     3361 tatacggagt gtataactgc ggctaagttg gcaattgaaa agtgtggtgc gaaaagtgta
     3421 ttgctgtcct ttgaaccaat gaatgatttg atgagccaaa aaaatgtaat caatattgtt
     3481 gatgataata ttaagaagta ccacacggaa tatacctaaa cggagccaat gtacgcaaaa
     3541 aaccccgctt cagcggggtt ttttcgccaa aaaaaacccc gccctgtcag gggcggggtt
     3601 tttttttagg gccctactga ctgtccgtgt gcttctcaaa tgcctgaggc cagtttgctc
     3661 aggctctccc cgtggaggta ataattgacg atatgatcag tgcacggcta actaagcggc
     3721 ctgctgactt tctcgccgat caaaaggcat tttgctatta agggattgac gagggcgtat
     3781 ctgcgcagta agatgcgccc cgcattccgg cggtagttca gcagggcaga acggcggact
     3841 ctaaatccgc atggcagggg ttcaaatccc ctccgccgga ccaaattcga aaagcctgct
     3901 caacgagcag gcttttttgc atgctcgagc agctcagggt cgaatttgct ttcgaatttc
     3961 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     4021 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     4081 cattaagcat tctgccgacg ttcatcggat cccgggcccg tcgactgcag aggcctgcat
     4141 gcaagcttgg cgtaatcatg atggacttaa ttaacggcac tgcgaaaaaa ccccgccgaa
     4201 gcggggtttt ttgcgaaata taatgaccct cttgataacc caagagggca ttttttaatg
     4261 cccatggcgt ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4321 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4381 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4441 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4501 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4561 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4621 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4681 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4741 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4801 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4861 gaccgagttg ctcttgccca gcgtcaatac gggataatac cgcgccacat agcagaactt
     4921 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4981 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5041 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5101 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5161 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5221 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     5281 ttatcatgac attaacctat aaaaataggc gtatcacgag gcccttaggc caaataggcc
     5341 gttcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg tttttgcgtg
     5401 agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa aaattttgcc
     5461 tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg
     5521 ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc
     5581 tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc
     5641 aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt
     5701 aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta
     5761 ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga
     5821 gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt
     5881 tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa
     5941 ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt
     6001 gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc
     6061 atcagctctc tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc
     6121 atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca
     6181 atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag
     6241 tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga catacatctc
     6301 aattggtcta ggtaatttta atcactatac caattgagat gggctagtca atgataatta
     6361 ctagtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa
     6421 cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt
     6481 tatattaaag tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata
     6541 gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg
     6601 tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc
     6661 accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccaacca tcaggcacct
     6721 gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg
     6781 gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac ccataataca
     6841 agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat
     6901 ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat
     6961 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa
     7021 caggcttacc cgtcttactg tcaagaggac atccggtcaa a
//
