LOCUS       sequence-314978-        3061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for cloning human 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 3061)
  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 3061)
  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-Human ABeta-attR5
FEATURES             Location/Qualifiers
     source          1..3061
                     /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    337..460
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(509..525)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(509..525)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(522..544)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    533..549
                     /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(557..563,564..581,582..587))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    602..623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(740..757)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(911..1499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(991..1010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1670..2485)
                     /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     2393..2412
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2486..2588)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2656..2674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2712..2734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2834..2853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3047..3061,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3061,1..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 ggcttaatgg atgcagaatt ccgacatgac tcaggatatg
      241 aagttcatca tcaaaaattg gtgttctttg cagaagatgt gggttcaaac aaaggtgcaa
      301 tcattggact catggtgggc ggtgttgtca tagcgacaac tttgtataca aaagttgaac
      361 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
      421 ctacataata ctgtaaaaca caacatatcc agtcactatg atcagttctg gaccagcgag
      481 ctgtgctgcg actcgtggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      541 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      601 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      661 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      721 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      781 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      841 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      901 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      961 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1021 ccctcgtgcg ctctcctgtt ccgaccctgt cgcttaccgg atacctgtcc gcctttctcc
     1081 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1141 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1201 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1261 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1321 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1381 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1441 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1501 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1561 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1621 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt tagaaaaact
     1681 catcgagcat caagtgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1741 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1801 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     1861 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     1921 agaatggcaa gagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     1981 cgtcatcaaa atcactcgca ccaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2041 gacgaaatac gcgatcgccg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2101 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2161 cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2221 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agcctgacca
     2281 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2341 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2401 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcaagcaag
     2461 acgtttcccg ttgaatatgg ctcattttag cttccttagc tcctgaaaat ctcgataact
     2521 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     2581 gccgatcaca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     2641 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     2701 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2761 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2821 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccaaat gcggtgtgaa
     2881 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     2941 cgcaactgtt gggaagggcg atcggtgcgg gcctcatcgc tattacgcca gctggcgaaa
     3001 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     3061 t
//
