LOCUS       sequence-454434-        3387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express A-beta-42-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3387)
  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 3387)
  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: pBL066c
FEATURES             Location/Qualifiers
     source          1..3387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     489..508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     742..759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    774..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    1404..1422
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1423..1477
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1428..1446
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1458..1476
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1704..2417
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1734..1762)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2357..2377
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(2495..3286)
                     /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(3287..3378)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(3387,1..588)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
       61 gcggtggttt gtttgccgga tcaagagcta ccacctcttt ttccgaaggt aactggcttc
      121 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
      781 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acaagaggac atccggtcaa
     1321 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttgctgagga gacttaggga
     1381 ccctccatcg aatggccaga tgatccctat cagtgataga gattgacact ctatcattga
     1441 tagagttatt ttaccactcc ctatcagtga tagagaaaag tgaaatgaat agttcgacaa
     1501 aaatctagac ctgcaggtgc agtaaggagg aaaaaaaaat ggacgcagag ttccgtcacg
     1561 acagtggcta tgaagtccac caccagaaat tggttttttt cgccgaggat gttggtagca
     1621 ataaaggggc tatcatcggt ttaatggtag gtggagttgt tattgcaggt tctgctggat
     1681 ctgctgctgg ttcgggggaa ttcatgagta aaggagaaga acttttcact ggagttgtcc
     1741 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
     1801 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
     1861 tacctgttcc atggccaaca cttgtcacta ctttaactta tggtgttcaa tgcttttcaa
     1921 gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgcct gaaggttatg
     1981 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
     2041 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
     2101 atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca
     2161 tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag
     2221 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     2281 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
     2341 aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca
     2401 tggatgaact atacaaataa attaacggca ctcctcagca aatataatga ccctcttgat
     2461 aacccaagag ggcatttttt aatgcccatg gcgtttattt gccgactacc ttggtgatct
     2521 cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt
     2581 cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca
     2641 ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc
     2701 tgtaccaaat gcgggacaac gtaagcacta catttcgctc atctccagcc cagtcgggcg
     2761 gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg
     2821 gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg
     2881 tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt
     2941 cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga
     3001 tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg
     3061 aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta
     3121 cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg
     3181 agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta
     3241 cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcatactc ttcctttttc
     3301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3361 tttagaaaaa taggccaaat aggccgt
//
