LOCUS       sequence-192214-        7034 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 7034)
  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 7034)
  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: pDB021CH(+)
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..99
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        80..98
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             136..1410
                     /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             1410..2489
                     /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             2553..3533
                     /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(4258..5049,5050..5118))
                     /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     4881..4900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5119..5223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5291..5309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5330..6280)
                     /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      6328..6550
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gacatccggt caaataaaac gaaaggctca gtcgaaagac ttggcctttc gttttgctga
       61 ggagacttag ggaccctact aatacgactc actataggga gaaagacctg caggtgcagt
      121 aaggaggaaa aaaaaatgaa aaaattatta ttcgcaattc ctttagttgt tcctttctat
      181 tctcactccg ctgaaactgt tgaaagttgt ttagcaaaac cccatacaga aaattcattt
      241 actaacgtct ggaaagacga caaaacttta gatcgttacg ctaactatga gggctgtctg
      301 tggaatgcta caggcgttgt agtttgtact ggtgacgaaa ctcagtgtta cggtacatgg
      361 gttcctattg ggcttgctat ccctgaaaat gagggtggtg gctctgaggg tggcggttct
      421 gagggtggcg gttctgaggg tggcggtact aaacctcctg agtacggtga tacacctatt
      481 ccgggctata cttatatcaa ccctctcgac ggcacttatc cgcctggtac tgagcaaaac
      541 cccgctaatc ctaatccttc tcttgaggag tctcagcctc ttaatacttt catgtttcag
      601 aataataggt tccgaaatag gcagggggca ttaactgttt atacgggcac tgttactcaa
      661 ggcactgacc ccgttaaaac ttattaccag tacactcctg tatcatcaaa agccatgtat
      721 gacgcttact ggaacggtaa attcagagac tgcgctttcc attctggctt taatgaggat
      781 ccattcgttt gtgaatatca aggccaatcg tctgacctgc ctcaacctcc tgtcaatgct
      841 ggcggcggct ctggtggtgg ttctggtggc ggctctgagg gtggtggctc tgagggtggc
      901 ggttctgagg gtggcggctc tgagggaggc ggttccggtg gtggctctgg ttccggtgat
      961 tttgattatg aaaagatggc aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa
     1021 aacgcgctac agtctgacgc taaaggcaaa cttgattctg tcgctactga ttacggtgct
     1081 gctatcgatg gtttcattgg tgacgtttcc ggccttgcta atggtaatgg tgctactggt
     1141 gattttgctg gctctaattc ccaaatggct caagtcggtg acggtgataa ttcaccttta
     1201 atgaataatt tccgtcaata tttaccttcc ctccctcaat cggttgaatg tcgccctttt
     1261 gtctttggcg ctggtaaacc ttacgagttc agtatcgact gcgataagat caacctgttc
     1321 cgcggtgtct ttgcgtttct tttatatgtt gccaccttta tgtatgtatt ttctacgttt
     1381 gctaacatac tgcgtaataa ggagtcttaa tgaaatttgg aaactttttg cttacatacc
     1441 aacctcccca attttcccaa acagaggtaa tgaaacgttt ggttaaatta ggtcgcatct
     1501 ctgaggagtg tggttttgat accgtatggt tactggagca tcatttcacg gagtttggtt
     1561 tgcttggtaa cccttatgtc gctgctgcat atttacttgg cgcgactaaa aaattgaatg
     1621 taggaactgc cgctattgtt cttcccacag cccatccagt acgccaactt gaagatgtga
     1681 atttattgga tcaaatgtca aaaggacgat ttcggtttgg tatttgccga gggctttaca
     1741 acaaggactt tcgcgtattc ggcacagata tgaataacag tcgcgcctta gcggaatgct
     1801 ggtacgggct gataaagaat ggcatgacag agggatatat ggaagctgat aatgaacata
     1861 tcaagttcca taaggtaaaa gtaaaccccg cggcgtatag cagaggtggc gcaccggttt
     1921 atgtggtggc tgaatcagct tcgacgactg agtgggctgc tcaatttggc ctaccgatga
     1981 tattaagttg gattataaat actaacgaaa agaaagcaca acttgagctt tataatgaag
     2041 tggctcaaga atatgggcac gatattcata atatcgacca ttgcttatca tatataacat
     2101 ctgtagatca tgactcaatt aaagcgaaag agatttgccg gaaatttctg gggcattggt
     2161 atgattctta tgtgaatgct acgactattt ttgatgattc agaccaaaca agaggttatg
     2221 atttcaataa agggcagtgg cgtgactttg tattaaaagg acataaagat actaatcgcc
     2281 gtattgatta cagttacgaa atcaatcccg tgggaacgcc gcaggaatgt attgacataa
     2341 ttcaaaaaga cattgatgct acaggaatat caaatatttg ttgtggattt gaagctaatg
     2401 gaacagtaga cgaaattatt gcttccatga agctcttcca gtctgatgtc atgccatttc
     2461 ttaaagaaaa acaacgttcg ctattatatt atggcggtgg cggtagcggc ggtggcggta
     2521 gcggcggtgg cggtagcggc ggtggcggta gcaaatttgg attgttcttc cttaacttca
     2581 tcaattcaac aactgttcaa gaacagagta tagttcgcat gcaggaaata acggagtatg
     2641 ttgataagtt gaattttgaa cagattttag tgtatgaaaa tcatttttca gataatggtg
     2701 ttgtcggcgc tcctctgact gtttctggtt ttctgctcgg tttaacagag aaaattaaaa
     2761 ttggttcatt aaatcacatc attacaactc atcatcctgt ccgcatagcg gaggaagctt
     2821 gcttattgga tcagttaagt gaagggagat ttattttagg gtttagtgat tgcgaaaaaa
     2881 aagatgaaat gcattttttt aatcgcccgg ttgaatatca acagcaacta tttgaagagt
     2941 gttatgaaat cattaacgat gctttaacaa caggctattg taatccagat aacgattttt
     3001 atagcttccc taaaatatct gtaaatcccc atgcttatac gccaggcgga cctcggaaat
     3061 atgtaacagc aaccagtcat catattgttg agtgggcggc caaaaaaggt attcctctca
     3121 tctttaagtg ggatgattct aatgatgtta gatatgaata tgctgaaaga tataaagccg
     3181 ttgcggataa atatgacgtt gacctatcag agatagacca tcagttaatg atattagtta
     3241 actataacga agatagtaat aaagctaaac aagagacgcg tgcatttatt agtgattatg
     3301 ttcttgaaat gcaccctaat gaaaatttcg aaaataaact tgaagaaata attgcagaaa
     3361 acgctgtcgg aaattatacg gagtgtataa ctgcggctaa gttggcaatt gaaaagtgtg
     3421 gtgcgaaaag tgtattgctg tcctttgaac caatgaatga tttgatgagc caaaaaaatg
     3481 taatcaatat tgttgatgat aatattaaga agtaccacac ggaatatacc taaacggagc
     3541 caatgtacgc aaaaaacccc gcttcagcgg ggttttttcg ccaaaaaaaa ccccgccctg
     3601 tcaggggcgg ggtttttttt tagggcccta ctgactgtcc gtgtgcttct caaatgcctg
     3661 aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga tcagtgcacg
     3721 gctaactaag cggcctgctg actttctcgc cgatcaaaag gcattttgct attaagggat
     3781 tgacgagggc gtatctgcgc agtaagatgc gccccgcatt cggaaacctg atcatgtaga
     3841 tcgaatggac tctaaatccg ttcagccggg ttagattccc ggggtttccg ccaggatccg
     3901 tcgacctgca gatccttagc gaaagctaag gatttttttt aagcttctgc cattcatccg
     3961 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     4021 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     4081 gccgacgttc atcggatccc gggcccgtcg actgcagagg cctgcatgca agcttggcgt
     4141 aatcatgatg gacttaatta acggcactgc gaaaaaaccc cgccgaagcg gggttttttg
     4201 cgaaatataa tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgttta
     4261 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4321 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4381 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4441 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4501 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4561 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4621 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4681 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4741 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4801 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4861 ttgcccagcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4921 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4981 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5041 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5101 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5161 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5221 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5281 aacctataaa aataggcgta tcacgaggcc cttaggccaa ataggccgtt cagatccttc
     5341 cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg
     5401 aaccattgag atcatgctta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga
     5461 gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta cgtaggtagg
     5521 aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa
     5581 gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg
     5641 ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta
     5701 ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta
     5761 acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg
     5821 ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa
     5881 catgttccag attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt
     5941 cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa
     6001 tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg
     6061 ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt
     6121 ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga
     6181 gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa
     6241 tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt
     6301 aattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc
     6361 tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg
     6421 ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attaaagtgg
     6481 ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg
     6541 tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt
     6601 ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct
     6661 cgggcaaatc gctgaatatt ccttttgtct ccaaccatca ggcacctgag tcgctgtctt
     6721 tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac
     6781 tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc
     6841 acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct
     6901 gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag
     6961 gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt
     7021 cttactgtca agag
//
