LOCUS       sequence-182087-        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     85..104
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        85..103
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             141..1415
                     /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             1415..2494
                     /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             2558..3538
                     /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(4290..5081,5082..5150))
                     /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     4913..4932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5151..5255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5323..5341
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5362..6312)
                     /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      6360..6582
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 aagaggacat ccggtcaaat aaaacgaaag gctcagtcga aagacttggc ctttcgtttt
       61 gctgaggaga cttagggacc ctactaatac gactcactat agggagaaag acctgcaggt
      121 gcagtaagga ggaaaaaaaa atgaaaaaat tattattcgc aattccttta gttgttcctt
      181 tctattctca ctccgctgaa actgttgaaa gttgtttagc aaaaccccat acagaaaatt
      241 catttactaa cgtctggaaa gacgacaaaa ctttagatcg ttacgctaac tatgagggct
      301 gtctgtggaa tgctacaggc gttgtagttt gtactggtga cgaaactcag tgttacggta
      361 catgggttcc tattgggctt gctatccctg aaaatgaggg tggtggctct gagggtggcg
      421 gttctgaggg tggcggttct gagggtggcg gtactaaacc tcctgagtac ggtgatacac
      481 ctattccggg ctatacttat atcaaccctc tcgacggcac ttatccgcct ggtactgagc
      541 aaaaccccgc taatcctaat ccttctcttg aggagtctca gcctcttaat actttcatgt
      601 ttcagaataa taggttccga aataggcagg gggcattaac tgtttatacg ggcactgtta
      661 ctcaaggcac tgaccccgtt aaaacttatt accagtacac tcctgtatca tcaaaagcca
      721 tgtatgacgc ttactggaac ggtaaattca gagactgcgc tttccattct ggctttaatg
      781 aggatccatt cgtttgtgaa tatcaaggcc aatcgtctga cctgcctcaa cctcctgtca
      841 atgctggcgg cggctctggt ggtggttctg gtggcggctc tgagggtggt ggctctgagg
      901 gtggcggttc tgagggtggc ggctctgagg gaggcggttc cggtggtggc tctggttccg
      961 gtgattttga ttatgaaaag atggcaaacg ctaataaggg ggctatgacc gaaaatgccg
     1021 atgaaaacgc gctacagtct gacgctaaag gcaaacttga ttctgtcgct actgattacg
     1081 gtgctgctat cgatggtttc attggtgacg tttccggcct tgctaatggt aatggtgcta
     1141 ctggtgattt tgctggctct aattcccaaa tggctcaagt cggtgacggt gataattcac
     1201 ctttaatgaa taatttccgt caatatttac cttccctccc tcaatcggtt gaatgtcgcc
     1261 cttttgtctt tggcgctggt aaaccttacg agttcagtat cgactgcgat aagatcaacc
     1321 tgttccgcgg tgtctttgcg tttcttttat atgttgccac ctttatgtat gtattttcta
     1381 cgtttgctaa catactgcgt aataaggagt cttaatgaaa tttggaaact ttttgcttac
     1441 ataccaacct ccccaatttt cccaaacaga ggtaatgaaa cgtttggtta aattaggtcg
     1501 catctctgag gagtgtggtt ttgataccgt atggttactg gagcatcatt tcacggagtt
     1561 tggtttgctt ggtaaccctt atgtcgctgc tgcatattta cttggcgcga ctaaaaaatt
     1621 gaatgtagga actgccgcta ttgttcttcc cacagcccat ccagtacgcc aacttgaaga
     1681 tgtgaattta ttggatcaaa tgtcaaaagg acgatttcgg tttggtattt gccgagggct
     1741 ttacaacaag gactttcgcg tattcggcac agatatgaat aacagtcgcg ccttagcgga
     1801 atgctggtac gggctgataa agaatggcat gacagaggga tatatggaag ctgataatga
     1861 acatatcaag ttccataagg taaaagtaaa ccccgcggcg tatagcagag gtggcgcacc
     1921 ggtttatgtg gtggctgaat cagcttcgac gactgagtgg gctgctcaat ttggcctacc
     1981 gatgatatta agttggatta taaatactaa cgaaaagaaa gcacaacttg agctttataa
     2041 tgaagtggct caagaatatg ggcacgatat tcataatatc gaccattgct tatcatatat
     2101 aacatctgta gatcatgact caattaaagc gaaagagatt tgccggaaat ttctggggca
     2161 ttggtatgat tcttatgtga atgctacgac tatttttgat gattcagacc aaacaagagg
     2221 ttatgatttc aataaagggc agtggcgtga ctttgtatta aaaggacata aagatactaa
     2281 tcgccgtatt gattacagtt acgaaatcaa tcccgtggga acgccgcagg aatgtattga
     2341 cataattcaa aaagacattg atgctacagg aatatcaaat atttgttgtg gatttgaagc
     2401 taatggaaca gtagacgaaa ttattgcttc catgaagctc ttccagtctg atgtcatgcc
     2461 atttcttaaa gaaaaacaac gttcgctatt atattatggc ggtggcggta gcggcggtgg
     2521 cggtagcggc ggtggcggta gcggcggtgg cggtagcaaa tttggattgt tcttccttaa
     2581 cttcatcaat tcaacaactg ttcaagaaca gagtatagtt cgcatgcagg aaataacgga
     2641 gtatgttgat aagttgaatt ttgaacagat tttagtgtat gaaaatcatt tttcagataa
     2701 tggtgttgtc ggcgctcctc tgactgtttc tggttttctg ctcggtttaa cagagaaaat
     2761 taaaattggt tcattaaatc acatcattac aactcatcat cctgtccgca tagcggagga
     2821 agcttgctta ttggatcagt taagtgaagg gagatttatt ttagggttta gtgattgcga
     2881 aaaaaaagat gaaatgcatt tttttaatcg cccggttgaa tatcaacagc aactatttga
     2941 agagtgttat gaaatcatta acgatgcttt aacaacaggc tattgtaatc cagataacga
     3001 tttttatagc ttccctaaaa tatctgtaaa tccccatgct tatacgccag gcggacctcg
     3061 gaaatatgta acagcaacca gtcatcatat tgttgagtgg gcggccaaaa aaggtattcc
     3121 tctcatcttt aagtgggatg attctaatga tgttagatat gaatatgctg aaagatataa
     3181 agccgttgcg gataaatatg acgttgacct atcagagata gaccatcagt taatgatatt
     3241 agttaactat aacgaagata gtaataaagc taaacaagag acgcgtgcat ttattagtga
     3301 ttatgttctt gaaatgcacc ctaatgaaaa tttcgaaaat aaacttgaag aaataattgc
     3361 agaaaacgct gtcggaaatt atacggagtg tataactgcg gctaagttgg caattgaaaa
     3421 gtgtggtgcg aaaagtgtat tgctgtcctt tgaaccaatg aatgatttga tgagccaaaa
     3481 aaatgtaatc aatattgttg atgataatat taagaagtac cacacggaat atacctaaac
     3541 ggagccaatg tacgcaaaaa accccgcttc agcggggttt tttcgccaaa aaaaaccccg
     3601 ccctgtcagg ggcggggttt ttttttaggg ccctactgac tgtccgtgtg cttctcaaat
     3661 gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga tatgatcagt
     3721 gcacggctaa ctaagcggcc tgctgacttt ctcgccgatc aaaaggcatt ttgctattaa
     3781 gggattgacg agggcgtatc tgcgcagtaa gatgcgcccc gcattccggc ggtagttcag
     3841 cagggcagaa cggcggactc taaatccgca tggcaggggt tcaaatcccc tccgccggac
     3901 caaattcgaa aagcctgctc aacgagcagg cttttttgca tgctcgagca gctcagggtc
     3961 gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcaa
     4021 ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
     4081 tcgcagtact gttgtaattc attaagcatt ctgccgacgt tcatcggatc ccgggcccgt
     4141 cgactgcaga ggcctgcatg caagcttggc gtaatcatga tggacttaat taacggcact
     4201 gcgaaaaaac cccgccgaag cggggttttt tgcgaaatat aatgaccctc ttgataaccc
     4261 aagagggcat tttttaatgc ccatggcgtt taccaatgct taatcagtga ggcacctatc
     4321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4381 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4441 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4501 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4801 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4861 tgagaatagt gtatgcggcg accgagttgc tcttgcccag cgtcaatacg ggataatacc
     4921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5041 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5101 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5161 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5221 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5281 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5341 cccttaggcc aaataggccg ttcagatcct tccgtattta gccagtatgt tctctagtgt
     5401 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat
     5461 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     5521 tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt
     5581 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     5641 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     5701 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     5761 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     5821 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     5881 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     5941 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     6001 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     6061 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     6121 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     6181 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     6241 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     6301 taattcagac atacatctca attggtctag gtaattttaa tcactatacc aattgagatg
     6361 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg
     6421 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     6481 tttgtgtgtt ttttttgttt atattaaagt ggttataatt tatagaataa agaaagaata
     6541 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     6601 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     6661 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt
     6721 ctccaaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
     6781 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
     6841 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg
     6901 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc
     6961 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt
     7021 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt c
//
