LOCUS       sequence-228467-        4735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the amyloid-beta protein (40 aa), containing an exogenous 
            methionine (Abeta(M1-40)), 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 4735)
  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 4735)
  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(M1-40)
FEATURES             Location/Qualifiers
     source          1..4735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(265..283)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      279..326
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(692..697,698..714,715..720))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(747..765)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        833..937
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(938..1006,1007..1798)
                     /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(1156..1175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1969..2557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2458..2477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2711..2728
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2743..2882
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2808..2827)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2968..2990
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2984..3175)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4633..4652
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atggacgctg aattccgtca cgactctggt tacgaagttc
      121 accaccagaa gctggtgttc ttcgctgaag acgtgggttc taacaagggt gctatcatcg
      181 gtctgatggt tggtggcgtt gtgtaatagg agctcgatcc ggctgctaac aaagcccgaa
      241 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      301 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg
      361 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg
      421 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc
      481 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg
      541 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
      601 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
      661 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
      721 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
      781 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
      841 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      901 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      961 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1021 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1081 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1141 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1201 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1261 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1321 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1381 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1441 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1501 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1561 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1621 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1681 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1741 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1801 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1861 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1921 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1981 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2041 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2101 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2161 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2221 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2281 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2341 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2401 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2461 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2521 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2581 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2641 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2701 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2761 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2821 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2881 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2941 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3001 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3061 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3121 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3181 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3241 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3301 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3361 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3421 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3481 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3541 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3601 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     3661 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     3721 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     3781 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     3841 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     3901 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     3961 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     4021 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     4081 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     4141 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     4201 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     4261 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     4321 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     4381 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg
     4441 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     4501 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     4561 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     4621 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     4681 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaat
//
