LOCUS       sequence-454429-        6526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementary plasmid providing gIII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6526)
  AUTHORS   Yang L, Zhang J, Andon JS, Li L, Wang T
  TITLE     Rapid discovery of cyclic peptide protein aggregation inhibitors by 
            continuous selection.
  JOURNAL   Nat Chem Biol. 2025 Jan 13. doi: 10.1038/s41589-024-01823-x.
  PUBMED    39806068
REFERENCE   2  (bases 1 to 6526)
  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. 2025 Jan 13. doi: 10.1038/s41589-024-01823-x."
COMMENT     Sequence Label: pLY034b
FEATURES             Location/Qualifiers
     source          1..6526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..951)
                     /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      999..1221
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1978..3252
                     /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             3252..4331
                     /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             4395..5375
                     /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(5455..6246,6247..6315))
                     /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     6078..6097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6316..6420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6488..6506
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcttactgtc aagaggacat ccggtcaaat aaaacgaaag gctcagtcga
     1741 aagactgggc ctttcgtttt gctgaggaga cttagggacc ctgatgaaat tcgccacttg
     1801 ttagtgtaat tcgctaactc atcctggcat gttgctgttg attcttcaat cagatcttta
     1861 taaatcaaaa agataaaaaa ttggcacgca aattgtatta acagttcagc aggacaatcc
     1921 tgaacgcaga aatcaagagg acaacattac ctgcaggtgc agtaaaaaag gaaaaaaatg
     1981 aaaaaattat tattcgcaat tcctttagtt gttcctttct attctcactc cgctgaaact
     2041 gttgaaagtt gtttagcaaa accccataca gaaaattcat ttactaacgt ctggaaagac
     2101 gacaaaactt tagatcgtta cgctaactat gagggctgtc tgtggaatgc tacaggcgtt
     2161 gtagtttgta ctggtgacga aactcagtgt tacggtacat gggttcctat tgggcttgct
     2221 atccctgaaa atgagggtgg tggctctgag ggtggcggtt ctgagggtgg cggttctgag
     2281 ggtggcggta ctaaacctcc tgagtacggt gatacaccta ttccgggcta tacttatatc
     2341 aaccctctcg acggcactta tccgcctggt actgagcaaa accccgctaa tcctaatcct
     2401 tctcttgagg agtctcagcc tcttaatact ttcatgtttc agaataatag gttccgaaat
     2461 aggcaggggg cattaactgt ttatacgggc actgttactc aaggcactga ccccgttaaa
     2521 acttattacc agtacactcc tgtatcatca aaagccatgt atgacgctta ctggaacggt
     2581 aaattcagag actgcgcttt ccattctggc tttaatgagg atccattcgt ttgtgaatat
     2641 caaggccaat cgtctgacct gcctcaacct cctgtcaatg ctggcggcgg ctctggtggt
     2701 ggttctggtg gcggctctga gggtggtggc tctgagggtg gcggttctga gggtggcggc
     2761 tctgagggag gcggttccgg tggtggctct ggttccggtg attttgatta tgaaaagatg
     2821 gcaaacgcta ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac
     2881 gctaaaggca aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt
     2941 ggtgacgttt ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat
     3001 tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa
     3061 tatttacctt ccctccctca atcggttgaa tgtcgccctt ttgtctttgg cgctggtaaa
     3121 ccttacgagt tcagtatcga ctgcgataag atcaacctgt tccgcggtgt ctttgcgttt
     3181 cttttatatg ttgccacctt tatgtatgta ttttctacgt ttgctaacat actgcgtaat
     3241 aaggagtctt aatgaaattt ggaaactttt tgcttacata ccaacctccc caattttccc
     3301 aaacagaggt aatgaaacgt ttggttaaat taggtcgcat ctctgaggag tgtggttttg
     3361 ataccgtatg gttactggag catcatttca cggagtttgg tttgcttggt aacccttatg
     3421 tcgctgctgc atatttactt ggcgcgacta aaaaattgaa tgtaggaact gccgctattg
     3481 ttcttcccac agcccatcca gtacgccaac ttgaagatgt gaatttattg gatcaaatgt
     3541 caaaaggacg atttcggttt ggtatttgcc gagggcttta caacaaggac tttcgcgtat
     3601 tcggcacaga tatgaataac agtcgcgcct tagcggaatg ctggtacggg ctgataaaga
     3661 atggcatgac agagggatat atggaagctg ataatgaaca tatcaagttc cataaggtaa
     3721 aagtaaaccc cgcggcgtat agcagaggtg gcgcaccggt ttatgtggtg gctgaatcag
     3781 cttcgacgac tgagtgggct gctcaatttg gcctaccgat gatattaagt tggattataa
     3841 atactaacga aaagaaagca caacttgagc tttataatga agtggctcaa gaatatgggc
     3901 acgatattca taatatcgac cattgcttat catatataac atctgtagat catgactcaa
     3961 ttaaagcgaa agagatttgc cggaaatttc tggggcattg gtatgattct tatgtgaatg
     4021 ctacgactat ttttgatgat tcagaccaaa caagaggtta tgatttcaat aaagggcagt
     4081 ggcgtgactt tgtattaaaa ggacataaag atactaatcg ccgtattgat tacagttacg
     4141 aaatcaatcc cgtgggaacg ccgcaggaat gtattgacat aattcaaaaa gacattgatg
     4201 ctacaggaat atcaaatatt tgttgtggat ttgaagctaa tggaacagta gacgaaatta
     4261 ttgcttccat gaagctcttc cagtctgatg tcatgccatt tcttaaagaa aaacaacgtt
     4321 cgctattata ttatggcggt ggcggtagcg gcggtggcgg tagcggcggt ggcggtagcg
     4381 gcggtggcgg tagcaaattt ggattgttct tccttaactt catcaattca acaactgttc
     4441 aagaacagag tatagttcgc atgcaggaaa taacggagta tgttgataag ttgaattttg
     4501 aacagatttt agtgtatgaa aatcattttt cagataatgg tgttgtcggc gctcctctga
     4561 ctgtttctgg ttttctgctc ggtttaacag agaaaattaa aattggttca ttaaatcaca
     4621 tcattacaac tcatcatcct gtccgcatag cggaggaagc ttgcttattg gatcagttaa
     4681 gtgaagggag atttatttta gggtttagtg attgcgaaaa aaaagatgaa atgcattttt
     4741 ttaatcgccc ggttgaatat caacagcaac tatttgaaga gtgttatgaa atcattaacg
     4801 atgctttaac aacaggctat tgtaatccag ataacgattt ttatagcttc cctaaaatat
     4861 ctgtaaatcc ccatgcttat acgccaggcg gacctcggaa atatgtaaca gcaaccagtc
     4921 atcatattgt tgagtgggcg gccaaaaaag gtattcctct catctttaag tgggatgatt
     4981 ctaatgatgt tagatatgaa tatgctgaaa gatataaagc cgttgcggat aaatatgacg
     5041 ttgacctatc agagatagac catcagttaa tgatattagt taactataac gaagatagta
     5101 ataaagctaa acaagagacg cgtgcattta ttagtgatta tgttcttgaa atgcacccta
     5161 atgaaaattt cgaaaataaa cttgaagaaa taattgcaga aaacgctgtc ggaaattata
     5221 cggagtgtat aactgcggct aagttggcaa ttgaaaagtg tggtgcgaaa agtgtattgc
     5281 tgtcctttga accaatgaat gatttgatga gccaaaaaaa tgtaatcaat attgttgatg
     5341 ataatattaa gaagtaccac acggaatata cctaaactta attaacggca ctcctcagca
     5401 aatataatga ccctcttgat aacccaagag ggcatttttt aatgcccatg gcgtttacca
     5461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6421 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6481 ctataaaaat aggcgtatca cgaggccctt aggccaaata ggccgt
//
