LOCUS       sequence-322446-        4741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the amyloid-beta protein (42 aa), containing an exogenous 
            methionine (Abeta(M1-42)), when expressed in BL21* DE3 p LysS E. 
            coli cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4741)
  AUTHORS   Walsh DM, Thulin E, Minogue AM, Gustavsson N, Pang E, Teplow DB, 
            Linse S
  TITLE     A facile method for expression and purification of the Alzheimer's 
            disease-associated amyloid beta-peptide.
  JOURNAL   FEBS J. 2009 Mar;276(5):1266-81. doi: 
            10.1111/j.1742-4658.2008.06862.x.
  PUBMED    19175671
REFERENCE   2  (bases 1 to 4741)
  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"; doi: 
            "10.1111/j.1742-4658.2008.06862"; journalName: "FEBS J"; date: 
            "2009-03"; volume: "276"; issue: "5"; pages: "1266-81"
COMMENT     Sequence Label: pET-Sac-Abeta(M1-42)
FEATURES             Location/Qualifiers
     source          1..4741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     375..394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     628..645
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    660..799
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(725..744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     885..907
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(901..1092)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2550..2569
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2653..2672
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2653..2671
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2703..2725
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2923..2941)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2937..2984
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3350..3355,3356..3372,3373..3378))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3405..3423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3491..3595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3596..3664,3665..4456)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3814..3833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4627..4741,1..474)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
       61 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      541 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      601 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      661 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
      721 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
      781 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      841 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
      901 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
      961 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     1021 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     1081 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     1141 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     1201 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     1261 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     1321 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     1381 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     1441 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     1501 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     1561 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
     1621 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc
     1681 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag
     1741 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg
     1801 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg
     1861 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt
     1921 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg
     1981 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc
     2041 agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg
     2101 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     2161 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     2221 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     2281 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     2341 aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     2401 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     2461 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     2521 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     2581 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     2641 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
     2701 attttgttta actttaagaa ggagatatac atatggacgc tgaattccgt cacgactctg
     2761 gttacgaagt tcaccaccag aagctggtgt tcttcgctga agacgtgggt tctaacaagg
     2821 gtgctatcat cggtctgatg gttggtggcg ttgtgatcgc ttaataggag ctcgatccgg
     2881 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     2941 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     3001 tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca
     3061 agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg
     3121 ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg
     3181 atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     3241 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
     3301 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
     3361 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
     3421 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3481 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3541 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3601 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3661 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3721 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3781 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     3841 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3901 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3961 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4021 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4081 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     4141 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4201 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4261 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4321 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4381 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4441 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4501 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4561 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4621 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4681 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4741 t
//
