LOCUS       sequence-210477-        7198 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 7198)
  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 7198)
  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: pDB026a
FEATURES             Location/Qualifiers
     source          1..7198
                     /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(4427..5218,5219..5287))
                     /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     5050..5069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5288..5392)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5460..5478
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5499..6449)
                     /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      6497..6719
                     /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 aacttattac 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 cgtagcacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
     4201 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacgttca
     4261 tcggatcccg ggcccgtcga ctgcagaggc ctgcatgcaa gcttggcgta atcatgatgg
     4321 acttaattaa cggcactgcg aaaaaacccc gccgaagcgg ggttttttgc gaaatataat
     4381 gaccctcttg ataacccaag agggcatttt ttaatgccca tggcgtttac caatgcttaa
     4441 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4501 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4561 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4621 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4681 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4741 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4801 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4861 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4921 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4981 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5041 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5101 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5161 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5221 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5281 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5341 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5401 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5461 ataggcgtat cacgaggccc ttaggccaaa taggccgttc agatccttcc gtatttagcc
     5521 agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga
     5581 tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt
     5641 gcagttaaag catcgtgtag tgtttttctt agtccgttac gtaggtagga atctgatgta
     5701 atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg
     5761 agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc
     5821 ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa
     5881 acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta
     5941 aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt
     6001 aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga
     6061 ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact
     6121 aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct
     6181 ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct
     6241 aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg
     6301 aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca
     6361 cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca
     6421 tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca
     6481 ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg
     6541 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
     6601 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
     6661 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
     6721 ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta
     6781 caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg
     6841 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
     6901 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
     6961 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
     7021 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
     7081 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
     7141 tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtc
//
