LOCUS       sequence-314980-        3118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for cloning nematode-codon-optimized Amyloid Beta(1-42)
            using 2-fragment gateway recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3118)
  AUTHORS   Kaur P, Kibat C, Teo E, Gruber J, Mathuru A, Tolwinski ANS
  TITLE     Use of Optogenetic Amyloid-beta to Monitor Protein Aggregation in 
            Drosophila melanogaster, Danio rerio and Caenorhabditis elegans.
  JOURNAL   Bio Protoc. 2020 Dec 5;10(23):e3856. doi: 10.21769/BioProtoc.3856. 
            eCollection 2020 Dec 5.
  PUBMED    33659494
REFERENCE   2  (bases 1 to 3118)
  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: "Bio 
            Protoc."; date: "2020-12-5"; volume: "10(23):e3856. doi"; pages: " 
            10.21769/BioProtoc.3856. eCollection 2020 Dec 5"
COMMENT     Sequence Label: pUCIDT-attL1-Worm ABeta-attR5
FEATURES             Location/Qualifiers
     source          1..3118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    105..204
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    394..517
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(566..582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(566..582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(579..601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    590..606
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(614..620,621..638,639..644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    659..680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(797..814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(968..1556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1048..1067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1727..2542)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2450..2469
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2543..2645)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2713..2731
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2769..2791)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2891..2910
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3104..3118,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3118,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgca acgcgatgac gatggatagc gattcatcga tgagctgacc
       61 cgatcgccgc cgccggaggg ttgcgtttga gacgggcgac agatcaaata atgattttat
      121 tttgactgat agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc
      181 caactttgta caaaaaagca ggcttaatgc ataaggtttt gctggcactg ttctttatct
      241 ttctggcacc agcaggaaca gatgctgatg cagaatttcg acatgattca ggatatgaag
      301 ttcatcatca aaaattggtg ttctttgcag aagatgtggg atcaaataaa ggagcaatta
      361 ttggactcat ggtgggagga gttgtcattg ctacaacttt gtatacaaaa gttgaacgag
      421 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
      481 cataatactg taaaacacaa catatccagt cactatgatc agttctggac cagcgagctg
      541 tgctgcgact cgtggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      601 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      661 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      781 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1081 tcgtgcgctc tcctgttccg accctgtcgc ttaccggata cctgtccgcc tttctccctt
     1141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1381 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat
     1741 cgagcatcaa gtgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     1801 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     1861 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     1921 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     1981 atggcaagag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     2041 catcaaaatc actcgcacca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     2101 gaaatacgcg atcgccgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     2161 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     2221 ggaatgctgt tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     2281 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagc ctgaccatct
     2341 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     2401 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     2461 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcttc aagcaagacg
     2521 tttcccgttg aatatggctc attttagctt ccttagctcc tgaaaatctc gataactcaa
     2581 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     2641 gatcacacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     2701 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     2761 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2821 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     2881 cttaactatg cggcatcaga gcagattgta ctgagagtgc accaaatgcg gtgtgaaata
     2941 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     3001 aactgttggg aagggcgatc ggtgcgggcc tcatcgctat tacgccagct ggcgaaaggg
     3061 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgtt
//
