LOCUS       sequence-454426-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low-stringency accessory plasmid for selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6453)
  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 6453)
  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: pTW357b
FEATURES             Location/Qualifiers
     source          1..6453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     292..311
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     545..562
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    577..717
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(643..662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     803..825
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(819..1010)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             1245..2519
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     terminator      complement(4943..4970)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5062..5148)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5301..5318
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(5364..6155)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(6156..6227)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6256..6453,1..391)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
       61 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      121 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      181 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      241 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      301 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      361 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      421 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      481 ggataaccgt attacagcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      541 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
      601 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
      661 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
      721 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtcttctccc ggcatccgct
      781 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
      841 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
      901 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
      961 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     1021 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     1081 acgatacggg ttactgatga tgaacaagag gacatccggt caaataaaac gaaaggctca
     1141 gtcgaaagac tgggcctttc gttttgctga ggagacttag ggaccctact aatacgtctc
     1201 actataggga gaaagacctg caggtgcagt aaaaaaggaa aaaaatgaaa aaattattat
     1261 tcgcaattcc tttagttgtt cctttctatt ctcactccgc tgaaactgtt gaaagttgtt
     1321 tagcaaaacc ccatacagaa aattcattta ctaacgtctg gaaagacgac aaaactttag
     1381 atcgttacgc taactatgag ggctgtctgt ggaatgctac aggcgttgta gtttgtactg
     1441 gtgacgaaac tcagtgttac ggtacatggg ttcctattgg gcttgctatc cctgaaaatg
     1501 agggtggtgg ctctgagggt ggcggttctg agggtggcgg ttctgagggt ggcggtacta
     1561 aacctcctga gtacggtgat acacctattc cgggctatac ttatatcaac cctctcgacg
     1621 gcacttatcc gcctggtact gagcaaaacc ccgctaatcc taatccttct cttgaggagt
     1681 ctcagcctct taatactttc atgtttcaga ataataggtt ccgaaatagg cagggggcat
     1741 taactgttta tacgggcact gttactcaag gcactgaccc cgttaaaact tattaccagt
     1801 acactcctgt atcatcaaaa gccatgtatg acgcttactg gaacggtaaa ttcagagact
     1861 gcgctttcca ttctggcttt aatgaggatc cattcgtttg tgaatatcaa ggccaatcgt
     1921 ctgacctgcc tcaacctcct gtcaatgctg gcggcggctc tggtggtggt tctggtggcg
     1981 gctctgaggg tggtggctct gagggtggcg gttctgaggg tggcggctct gagggaggcg
     2041 gttccggtgg tggctctggt tccggtgatt ttgattatga aaagatggca aacgctaata
     2101 agggggctat gaccgaaaat gccgatgaaa acgcgctaca gtctgacgct aaaggcaaac
     2161 ttgattctgt cgctactgat tacggtgctg ctatcgatgg tttcattggt gacgtttccg
     2221 gccttgctaa tggtaatggt gctactggtg attttgctgg ctctaattcc caaatggctc
     2281 aagtcggtga cggtgataat tcacctttaa tgaataattt ccgtcaatat ttaccttccc
     2341 tccctcaatc ggttgaatgt cgcccttttg tctttggcgc tggtaaacct tacgagttca
     2401 gtatcgactg cgataagatc aacctgttcc gcggtgtctt tgcgtttctt ttatatgttg
     2461 ccacctttat gtatgtattt tctacgtttg ctaacatact gcgtaataag gagtcttaaa
     2521 cggagccaat gtacgcaaaa aaccccgctt cagcggggtt ttttcgccaa aaaaaacccc
     2581 gccctgtcag gggcggggtt tttttttagg gccctactga ctgttacgcg aacgcgaagt
     2641 ccgactctaa gatgtcacgg aggttcaagt tacctttagc cggaagtgct ggcattttgt
     2701 ccaattgaga ctcgtgcaac tggtcagcga actggtcgta gaaatcagcc agtacatcac
     2761 aagactcata tgtgtcaacc atagtttcgc gcactgcttt gaacaggttc gcagcgtcag
     2821 ccggaatggt accgaaggag tcgtgaatca gtgcaaaaga ttcgattccg tacttctcgt
     2881 gtgcccacac tacagtctta cgaaggtggc taccatcttg gctgtgtaca aagttaggag
     2941 cgataccaga ctcctgtttg tgtgcatcaa tctcgctatc tttgttggtg ttaatggtag
     3001 gctgtaagcg gaactgaccg aggaacatca ggttcaagcg cgtctgaata ggcttcttgt
     3061 attcctgcca cacagggaaa ccatcaggag ttacccaatg cacagcgcaa cgcttgcgaa
     3121 gaatctctcc agtcttctta tctttgacct cagcagccag cagcttagca gcagacttaa
     3181 gccagttcat tgcttcaacc gcagctacca ccgtcacgct cacagattcc caaatcagct
     3241 tagccatgta tccagcagcc tgattcggct gagtgaacat cagacccttg ccggaatcaa
     3301 tagctggctg aatggtatct tccagcactt gttgacggaa gccgaactct ttggacccgt
     3361 aagccagcgt catgactgaa ctcttagtca cactgcgagt aacaccgtaa gccagccatt
     3421 gaccagccag tgccttagtg cccagcttga ctttctcaga gatttcacca gtgttctcat
     3481 cggtcacggt aactacttcg ttatcggtcc cattgattgc gtctgcttgt agaatctcgt
     3541 tgactttctt agcaacaatc ccgtagatgt cctgaacggt ttcactagga agcaagttaa
     3601 ccgcgcgacc acctacctca tctcggagca tcgcggagaa gtgctggatg ccagagcaag
     3661 acccgtcaaa cgccagcgga agggagcagt tatagctcag gccgtggtgc tgtaccccag
     3721 cgtactcaaa gcagaacgca aggaagcaga acggagaatc ttgctcagcc caccaagtgt
     3781 tctccagtgg agacttagcg caagccatga tgttctcgtg gttttcctca atgaacttga
     3841 tgcgctcagg gaacggaacc ttatcgacac ccgcacagtt tgcaccgtgg attttcagcc
     3901 agtagtaacc ttccttaccg attggtttac ctttcgccag cgtaagcagt cctttggtca
     3961 tatcgttacc ttgcgggttg aacattgaca cagcgtaaac acgaccgcgc cagtccatgt
     4021 tgtaagggaa ccagatggcc ttatggttag caaacttatt ggcttgctca agcatgaact
     4081 caaggctgat acggcgagac ttgcgagcct tgtccttgcg gtacacagca gcggcagcac
     4141 gtttccacgc ggtgagagcc tcaggattca tgtcgatgtc ttccggtttc atcgggagtt
     4201 cttcacgctc aatcgcaggg atgtcctcga ccggacaatg cttccacttg gtgattacgt
     4261 tggcgaccgc taggactttc ttgttgattt tccatgcggt gttttgcgca atgttaatcg
     4321 ctttgtacac ctcaggcatg taaacgtctt cgtagcgcat cagtgctttc ttactgtgag
     4381 tacgcaccag cgccagagga cgacgaccgt tagcccaata gccaccacca gtaatgccag
     4441 tccacggctt aggaggaact acgcaaggtt ggaacatcgg agagatgcca gccagcgcac
     4501 ctgcacgggt tgcgatagcc tcagcgtatt caggtgcgag ttcgatagtc tcagagtctt
     4561 gacctactac gccagcattt tggcggtgta agctaaccat tccggttgac tcaatgagca
     4621 tctcgatgca gcgtactcct acatgaatag agtcttcctt atgccacgaa gaccacgcct
     4681 cgccaccgag tagaccctta gagagcatgt cagcctcgac aacttgcata aatgctttca
     4741 tttttttttc ctccttactc tagtaaaagt taaacaaaat tatttgtaga gggaaaccgt
     4801 tgtggtctcc ctgaatatag ataccgagcc ttatgcatgc ccgtaaagtt atccagcaac
     4861 cactcataga cctagggcag cagataggga cgacgtggtg ttagctgtgc tctagaatcc
     4921 tcagcgagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttaatttg
     4981 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcgcaac
     5041 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     5101 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggcagt
     5161 tccctactct cgcatgggga gaccccacac taccatcggc gctacggcgt ttcacttctg
     5221 agttcggcat ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa ttctgtttta
     5281 tcagaccgct tctgcgttct gatttaatct gtatcaggct gaaaatcttc tctcatccgc
     5341 caaaacagcc atgcccatgg cgtttatttg ccgactacct tggtgatctc gcctttcacg
     5401 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
     5461 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     5521 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     5581 cgggacaacg taagcactac atttcgctca tctccagccc agtcgggcgg cgagttccat
     5641 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     5701 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     5761 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     5821 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     5881 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     5941 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     6001 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     6061 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     6121 tgagtcgata cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga
     6181 atcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6241 aggccaaata ggccgttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccac ctctttttcc
     6361 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6421 gttaggccac cacttcaaga actctgtagc acc
//
