LOCUS       sequence-380127-        4809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for human APP isoform having 751 amino acid residues.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4809)
  AUTHORS   Park H, Hundley FV, Yu Q, Overmyer KA, Brademan DR, Serrano L, Paulo
            JA, Paoli JC, Swarup S, Coon JJ, Gygi SP, Wade Harper J
  TITLE     Spatial snapshots of amyloid precursor protein intramembrane 
            processing via early endosome proteomics.
  JOURNAL   Nat Commun. 2022 Oct 16;13(1):6112. doi: 10.1038/s41467-022-33881-x.
  PUBMED    36245040
REFERENCE   2  (bases 1 to 4809)
  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 
            Commun."; date: "2022-10-16"; volume: "13(1):6112. doi"; pages: " 
            10.1038/s41467-022-33881-x"
COMMENT     Sequence Label: pENTR221-APP751
FEATURES             Location/Qualifiers
     source          1..4809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(125..152)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(244..330)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     379..401
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     393..410
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     394..410
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    426..525
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(858..869)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     CDS             1349..1369
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    complement(2791..2890)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2907..2926)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2908..2926)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2931..2947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2931..2947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3049..3068)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3060..3869
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3127..3146)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4016..4604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4505..4524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4758..4775
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
      121 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
      181 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
      241 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      301 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      361 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      421 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
      481 agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccacc atgctgcccg
      541 gtttggcact gctcctgctg gccgcctgga cggctcgggc gctggaggta cccactgatg
      601 gtaatgctgg cctgctggct gaaccccaga ttgccatgtt ctgtggcaga ctgaacatgc
      661 acatgaatgt ccagaatggg aagtgggatt cagatccatc agggaccaaa acctgcattg
      721 ataccaagga aggcatcctg cagtattgcc aagaagtcta ccctgaactg cagatcacca
      781 atgtggtaga agccaaccaa ccagtgacca tccagaactg gtgcaagcgg ggccgcaagc
      841 agtgcaagac ccatccccac tttgtgattc cctaccgctg cttagttggt gagtttgtaa
      901 gtgatgccct tctcgttcct gacaagtgca aattcttaca ccaggagagg atggatgttt
      961 gcgaaactca tcttcactgg cacaccgtcg ccaaagagac atgcagtgag aagagtacca
     1021 acttgcatga ctacggcatg ttgctgccct gcggaattga caagttccga ggggtagagt
     1081 ttgtgtgttg cccactggct gaagaaagtg acaatgtgga ttctgctgat gcggaggagg
     1141 atgactcgga tgtctggtgg ggcggagcag acacagacta tgcagatggg agtgaagaca
     1201 aagtagtaga agtagcagag gaggaagaag tggctgaggt ggaagaagaa gaagccgatg
     1261 atgacgagga cgatgaggat ggtgatgagg tagaggaaga ggctgaggaa ccctacgaag
     1321 aagccacaga gagaaccacc agcattgcca ccaccaccac caccaccaca gagtctgtgg
     1381 aagaggtggt tcgagaggtg tgctctgaac aagccgagac ggggccgtgc cgagcaatga
     1441 tctcccgctg gtactttgat gtgactgaag ggaagtgtgc cccattcttt tacggcggat
     1501 gtggcggcaa ccggaacaac tttgacacag aagagtactg catggccgtg tgtggcagcg
     1561 ccattcctac aacagcagcc agtacccctg atgccgttga caagtatctc gagacacctg
     1621 gggatgagaa tgaacatgcc catttccaga aagccaaaga gaggcttgag gccaagcacc
     1681 gagagagaat gtcccaggtc atgagagaat gggaagaggc agaacgtcaa gcaaagaact
     1741 tgcctaaagc tgataagaag gcagttatcc agcatttcca ggagaaagtg gaatctttgg
     1801 aacaggaagc agccaacgag agacagcagc tggtggagac acacatggcc agagtggaag
     1861 ccatgctcaa tgaccgccgc cgcctggccc tggagaacta catcaccgct ctgcaggctg
     1921 ttcctcctcg gcctcgtcac gtgttcaata tgctaaagaa gtatgtccgc gcagaacaga
     1981 aggacagaca gcacacccta aagcatttcg agcatgtgcg catggtggat cccaagaaag
     2041 ccgctcagat ccggtcccag gttatgacac acctccgtgt gatttatgag cgcatgaatc
     2101 agtctctctc cctgctctac aacgtgcctg cagtggccga ggagattcag gatgaagttg
     2161 atgagctgct tcagaaagag caaaactatt cagatgacgt cttggccaac atgattagtg
     2221 aaccaaggat cagttacgga aacgatgctc tcatgccatc tttgaccgaa acgaaaacca
     2281 ccgtggagct ccttcccgtg aatggagagt tcagcctgga cgatctccag ccgtggcatt
     2341 cttttggggc tgactctgtg ccagccaaca cagaaaacga agttgagcct gttgatgccc
     2401 gccctgctgc cgaccgagga ctgaccactc gaccaggttc tgggttgaca aatatcaaga
     2461 cggaggagat ctctgaagtg aagatggatg cagaattccg acatgactca ggatatgaag
     2521 ttcatcatca aaaattggtg ttctttgcag aagatgtggg ttcaaacaaa ggtgcaatca
     2581 ttggactcat ggtgggcggt gttgtcatag cgacagtgat cgtcatcacc ttggtgatgc
     2641 tgaagaagaa acagtacaca tccattcatc atggtgtggt ggaggttgac gccgctgtca
     2701 ccccagagga gcgccacctg tccaagatgc agcagaacgg ctacgaaaat ccaacctaca
     2761 agttctttga gcagatgcag aactagatcc acccagcttt cttgtacaaa gttggcatta
     2821 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     2881 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     2941 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     3001 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     3061 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     3121 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     3181 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     3241 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     3301 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3361 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3421 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3481 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3541 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3601 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3661 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3721 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3781 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3841 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     3901 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     3961 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4021 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4081 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4141 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4201 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4261 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4321 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4381 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4441 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4501 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4561 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4621 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4681 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4741 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4801 aaccgcctc
//
