LOCUS       sequence-210479-        7199 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 7199)
  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 7199)
  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: pDB026e
FEATURES             Location/Qualifiers
     source          1..7199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1553..2632
                     /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             2696..3676
                     /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(4428..5219,5220..5288))
                     /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     5051..5070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5289..5393)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5461..5479
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5500..6450)
                     /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      6498..6720
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 aagaggacat ccggtcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
       61 gctgaggaga cttagggacc ctgatgaaat tcgccacttg ttagtgtaat tcgctaactc
      121 atcctggcat gttgctgttg attcttcaat cagatcttta taaatcaaaa agataaaaaa
      181 ttggcacgca aattgtatta acagttcagc aggacaatcc tgaacgcaga aatcaagagg
      241 acaacattac ctgcaggtgc agtaaggagg aaaaaaaaat gaaaaaatta ttattcgcaa
      301 ttcctttagt tgttcctttc tattctcact ccgctgaaac tgttgaaagt tgtttagcaa
      361 aatagcatac agaaaattca tttactaacg tctggaaaga cgacaaaact ttagatcgtt
      421 acgctaacta tgagggctgt ctgtggaatg ctacaggcgt tgtagtttgt actggtgacg
      481 aaactcagtg ttacggtaca tgggttccta ttgggcttgc tatccctgaa aatgagggtg
      541 gtggctctga gggtggcggt tctgagggtg gcggttctga gggtggcggt actaaacctc
      601 ctgagtacgg tgatacacct attccgggct atacttatat caaccctctc gacggcactt
      661 atccgcctgg tactgagcaa aaccccgcta atcctaatcc ttctcttgag gagtctcagc
      721 ctcttaatac tttcatgttt cagaataata ggttccgaaa taggcagggg gcattaactg
      781 tttatacggg cactgttact caaggcactg accccgttaa aacttattag cagtacactc
      841 ctgtatcatc aaaagccatg tatgacgctt actggaacgg taaattcaga gactgcgctt
      901 tccattctgg ctttaatgag gatccattcg tttgtgaata tcaaggccaa tcgtctgacc
      961 tgcctcaacc tcctgtcaat gctggcggcg gctctggtgg tggttctggt ggcggctctg
     1021 agggtggtgg ctctgagggt ggcggttctg agggtggcgg ctctgaggga ggcggttccg
     1081 gtggtggctc tggttccggt gattttgatt atgaaaagat ggcaaacgct aataaggggg
     1141 ctatgaccga aaatgccgat gaaaacgcgc tacagtctga cgctaaaggc aaacttgatt
     1201 ctgtcgctac tgattacggt gctgctatcg atggtttcat tggtgacgtt tccggccttg
     1261 ctaatggtaa tggtgctact ggtgattttg ctggctctaa ttcccaaatg gctcaagtcg
     1321 gtgacggtga taattcacct ttaatgaata atttccgtca atatttacct tccctccctc
     1381 aatcggttga atgtcgccct tttgtctttg gcgctggtaa accttacgag ttcagtatcg
     1441 actgcgataa gatcaacctg ttccgcggtg tctttgcgtt tcttttatat gttgccacct
     1501 ttatgtatgt attttctacg tttgctaaca tactgcgtaa taaggagtct taatgaaatt
     1561 tggaaacttt ttgcttacat accaacctcc ccaattttcc caaacagagg taatgaaacg
     1621 tttggttaaa ttaggtcgca tctctgagga gtgtggtttt gataccgtat ggttactgga
     1681 gcatcatttc acggagtttg gtttgcttgg taacccttat gtcgctgctg catatttact
     1741 tggcgcgact aaaaaattga atgtaggaac tgccgctatt gttcttccca cagcccatcc
     1801 agtacgccaa cttgaagatg tgaatttatt ggatcaaatg tcaaaaggac gatttcggtt
     1861 tggtatttgc cgagggcttt acaacaagga ctttcgcgta ttcggcacag atatgaataa
     1921 cagtcgcgcc ttagcggaat gctggtacgg gctgataaag aatggcatga cagagggata
     1981 tatggaagct gataatgaac atatcaagtt ccataaggta aaagtaaacc ccgcggcgta
     2041 tagcagaggt ggcgcaccgg tttatgtggt ggctgaatca gcttcgacga ctgagtgggc
     2101 tgctcaattt ggcctaccga tgatattaag ttggattata aatactaacg aaaagaaagc
     2161 acaacttgag ctttataatg aagtggctca agaatatggg cacgatattc ataatatcga
     2221 ccattgctta tcatatataa catctgtaga tcatgactca attaaagcga aagagatttg
     2281 ccggaaattt ctggggcatt ggtatgattc ttatgtgaat gctacgacta tttttgatga
     2341 ttcagaccaa acaagaggtt atgatttcaa taaagggcag tggcgtgact ttgtattaaa
     2401 aggacataaa gatactaatc gccgtattga ttacagttac gaaatcaatc ccgtgggaac
     2461 gccgcaggaa tgtattgaca taattcaaaa agacattgat gctacaggaa tatcaaatat
     2521 ttgttgtgga tttgaagcta atggaacagt agacgaaatt attgcttcca tgaagctctt
     2581 ccagtctgat gtcatgccat ttcttaaaga aaaacaacgt tcgctattat attatggcgg
     2641 tggcggtagc ggcggtggcg gtagcggcgg tggcggtagc ggcggtggcg gtagcaaatt
     2701 tggattgttc ttccttaact tcatcaattc aacaactgtt caagaacaga gtatagttcg
     2761 catgcaggaa ataacggagt atgttgataa gttgaatttt gaacagattt tagtgtatga
     2821 aaatcatttt tcagataatg gtgttgtcgg cgctcctctg actgtttctg gttttctgct
     2881 cggtttaaca gagaaaatta aaattggttc attaaatcac atcattacaa ctcatcatcc
     2941 tgtccgcata gcggaggaag cttgcttatt ggatcagtta agtgaaggga gatttatttt
     3001 agggtttagt gattgcgaaa aaaaagatga aatgcatttt tttaatcgcc cggttgaata
     3061 tcaacagcaa ctatttgaag agtgttatga aatcattaac gatgctttaa caacaggcta
     3121 ttgtaatcca gataacgatt tttatagctt ccctaaaata tctgtaaatc cccatgctta
     3181 tacgccaggc ggacctcgga aatatgtaac agcaaccagt catcatattg ttgagtgggc
     3241 ggccaaaaaa ggtattcctc tcatctttaa gtgggatgat tctaatgatg ttagatatga
     3301 atatgctgaa agatataaag ccgttgcgga taaatatgac gttgacctat cagagataga
     3361 ccatcagtta atgatattag ttaactataa cgaagatagt aataaagcta aacaagagac
     3421 gcgtgcattt attagtgatt atgttcttga aatgcaccct aatgaaaatt tcgaaaataa
     3481 acttgaagaa ataattgcag aaaacgctgt cggaaattat acggagtgta taactgcggc
     3541 taagttggca attgaaaagt gtggtgcgaa aagtgtattg ctgtcctttg aaccaatgaa
     3601 tgatttgatg agccaaaaaa atgtaatcaa tattgttgat gataatatta agaagtacca
     3661 cacggaatat acctaaacgg agccaatgta cgcaaaaaac cccgcttcag cggggttttt
     3721 tcgccaaaaa aaaccccgcc ctgtcagggg cggggttttt ttttagggcc ctactgactg
     3781 tccgtgtgct tctcaaatgc ctgaggccag tttgctcagg ctctccccgt ggaggtaata
     3841 attgacgata tgatcagtgc acggctaact aagcggcctg ctgactttct cgccgatcaa
     3901 aaggcatttt gctattaagg gattgacgag ggcgtatctg cgcagtaaga tgcgccccgc
     3961 attccggcgg tagttcagca gggcagaacg gcggactcta aatccgcatg gcaggggttc
     4021 aaatcccctc cgccggacca aattcgaaaa gcctgctcaa cgagcaggct tttttgcatg
     4081 ctcgagcagc tcagggtcga atttgctttc gaatttctgc cattcatccg cttattatca
     4141 cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac
     4201 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacgttc
     4261 atcggatccc gggcccgtcg actgcagagg cctgcatgca agcttggcgt aatcatgatg
     4321 gacttaatta acggcactgc gaaaaaaccc cgccgaagcg gggttttttg cgaaatataa
     4381 tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgttta ccaatgctta
     4441 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4501 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4561 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4621 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4681 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4741 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4801 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4861 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4921 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4981 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccagcg
     5041 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5101 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5161 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5221 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5281 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5341 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5401 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5461 aataggcgta tcacgaggcc cttaggccaa ataggccgtt cagatccttc cgtatttagc
     5521 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     5581 atcatgctta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     5641 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta cgtaggtagg aatctgatgt
     5701 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     5761 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     5821 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     5881 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     5941 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     6001 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     6061 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     6121 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     6181 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     6241 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     6301 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     6361 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     6421 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     6481 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     6541 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     6601 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     6661 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     6721 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     6781 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     6841 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     6901 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     6961 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     7021 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     7081 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     7141 ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtc
//
