LOCUS       sequence-182095-        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     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(4263..5054,5055..5123))
                     /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     4886..4905
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5124..5228)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5296..5314
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(5335..6285)
                     /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      6333..6555
                     /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 gcattcggaa acctgatcat
     3841 gtagatcgaa tggactctaa atccgttcag ccgggttaga ttcccggggt ttccgccagg
     3901 atccgtcgac ctgcagatcc ttagcgaaag ctaaggattt tttttaagct tctgccattc
     3961 atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca ccaataactg
     4021 ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     4081 attctgccga cgttcatcgg atcccgggcc cgtcgactgc agaggcctgc atgcaagctt
     4141 ggcgtaatca tgatggactt aattaacggc actgcgaaaa aaccccgccg aagcggggtt
     4201 ttttgcgaaa tataatgacc ctcttgataa cccaagaggg cattttttaa tgcccatggc
     4261 gtttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4321 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4381 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4441 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4501 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4561 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4621 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4681 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4741 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4801 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4861 tgctcttgcc cagcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4921 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4981 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5041 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5101 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5161 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5221 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5281 acattaacct ataaaaatag gcgtatcacg aggcccttag gccaaatagg ccgttcagat
     5341 ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga
     5401 gaacgaacca ttgagatcat gcttactttg catgtcactc aaaaattttg cctcaaaact
     5461 ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttacgtag
     5521 gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt
     5581 ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc
     5641 agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt
     5701 acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag
     5761 cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt
     5821 ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga
     5881 cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat
     5941 ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg
     6001 gaaaatctca aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc
     6061 tctggttgct ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc
     6121 gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg
     6181 gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg
     6241 actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc
     6301 taggtaattt taatcactat accaattgag atgggctagt caatgataat tactagtcct
     6361 tttcctttga gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa
     6421 ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattaa
     6481 agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag
     6541 ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac
     6601 gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc
     6661 aagctcgggc aaatcgctga atattccttt tgtctccaac catcaggcac ctgagtcgct
     6721 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat
     6781 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc
     6841 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt
     6901 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt
     6961 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta
     7021 cccgtcttac tgtc
//
