LOCUS       sequence-237938-        6815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human APP fused to EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6815)
  AUTHORS   Currinn H, Guscott B, Balklava Z, Rothnie A, Wassmer T
  TITLE     APP controls the formation of PI(3,5)P vesicles through its binding 
            of the PIKfyve complex.
  JOURNAL   Cell Mol Life Sci. 2015 Jul 28.
  PUBMED    26216398
REFERENCE   2  (bases 1 to 6815)
  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: "Cell Mol 
            Life Sci. 2015 Jul 28."
COMMENT     Sequence Label: pEGFP-n1-APP
FEATURES             Location/Qualifiers
     source          1..6815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(961..972)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     CDS             1452..1472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      2755..2764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2761..2763,2764..2766,2767..3480)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2761..2763,2764..2766,2767..3477)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2806..2827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3067..3086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3414..3435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3603..3724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3640..3659)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3694..3713
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3731..4186)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3868..3889)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4080..4099
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4213..4317
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4319..4676
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4319..4510
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4342..4362)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4527..4662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4711..5505
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4711..5505
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4765..4784)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5375..5394
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5693..5712)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5737..5784
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6113..6701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6602..6621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttgcc accatgctgc ccggtttggc actgctcctg
      661 ctggccgcct ggacggctcg ggcgctggag gtacccactg atggtaatgc tggcctgctg
      721 gctgaacccc agattgccat gttctgtggc agactgaaca tgcacatgaa tgtccagaat
      781 gggaagtggg attcagatcc atcagggacc aaaacctgca ttgataccaa ggaaggcatc
      841 ctgcagtatt gccaagaagt ctaccctgaa ctgcagatca ccaatgtggt agaagccaac
      901 caaccagtga ccatccagaa ctggtgcaag cggggccgca agcagtgcaa gacccatccc
      961 cactttgtga ttccctaccg ctgcttagtt ggtgagtttg taagtgatgc ccttctcgtt
     1021 cctgacaagt gcaaattctt acaccaggag aggatggatg tttgcgaaac tcatcttcac
     1081 tggcacaccg tcgccaaaga gacatgcagt gagaagagta ccaacttgca tgactacggc
     1141 atgttgctgc cctgcggaat tgacaagttc cgaggggtag agtttgtgtg ttgcccactg
     1201 gctgaagaaa gtgacaatgt ggattctgct gatgcggagg aggatgactc ggatgtctgg
     1261 tggggcggag cagacacaga ctatgcagat gggagtgaag acaaagtagt agaagtagca
     1321 gaggaggaag aagtggctga ggtggaagaa gaagaagccg atgatgacga ggacgatgag
     1381 gatggtgatg aggtagagga agaggctgag gaaccctacg aagaagccac agagagaacc
     1441 accagcattg ccaccaccac caccaccacc acagagtctg tggaagaggt ggttcgagtt
     1501 cctacaacag cagccagtac ccctgatgcc gttgacaagt atctcgagac acctggggat
     1561 gagaatgaac atgcccattt ccagaaagcc aaagagaggc ttgaggccaa gcaccgagag
     1621 agaatgtccc aggtcatgag agaatgggaa gaggcagaac gtcaagcaaa gaacttgcct
     1681 aaagctgata agaaggcagt tatccagcat ttccaggaga aagtggaatc tttggaacag
     1741 gaagcagcca acgagagaca gcagctggtg gagacacaca tggccagagt ggaagccatg
     1801 ctcaatgacc gccgccgcct ggccctggag aactacatca ccgctctgca ggctgttcct
     1861 cctcggcctc gtcacgtgtt caatatgcta aagaagtatg tccgcgcaga acagaaggac
     1921 agacagcaca ccctaaagca tttcgagcat gtgcgcatgg tggatcccaa gaaagccgct
     1981 cagatccggt cccaggttat gacacacctc cgtgtgattt atgagcgcat gaatcagtct
     2041 ctctccctgc tctacaacgt gcctgcagtg gccgaggaga ttcaggatga agttgatgag
     2101 ctgcttcaga aagagcaaaa ctattcagat gacgtcttgg ccaacatgat tagtgaacca
     2161 aggatcagtt acggaaacga tgctctcatg ccatctttga ccgaaacgaa aaccaccgtg
     2221 gagctccttc ccgtgaatgg agagttcagc ctggacgatc tccagccgtg gcattctttt
     2281 ggggctgact ctgtgccagc caacacagaa aacgaagttg agcctgttga tgcccgccct
     2341 gctgccgacc gaggactgac cactcgacca ggttctgggt tgacaaatat caagacggag
     2401 gagatctctg aagtgaagat ggatgcagaa ttccgacatg actcaggata tgaagttcat
     2461 catcaaaaat tggtgttctt tgcagaagat gtgggttcaa acaaaggtgc aatcattgga
     2521 ctcatggtgg gcggtgttgt catagcgaca gtgatcgtca tcaccttggt gatgctgaag
     2581 aagaaacagt acacatccat tcatcatggt gtggtggagg ttgacgccgc tgtcacccca
     2641 gaggagcgcc acctgtccaa gatgcagcag aacggctacg aaaatccaac ctacaagttc
     2701 tttgagcaga tgcagaacgg gtcgacggta ccgcgggccc gggatccacc ggtcgccacc
     2761 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2821 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2881 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2941 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3001 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3061 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3121 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3181 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3241 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3301 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3361 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3421 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     3481 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3541 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3601 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3661 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3721 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3781 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3841 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3901 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3961 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4021 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4081 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4141 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4321 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4381 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4561 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4621 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4681 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4741 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4801 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4861 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4921 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4981 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5041 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5101 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5161 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5221 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5281 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5341 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5401 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5461 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5521 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5581 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5641 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5701 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5761 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5821 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5881 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5941 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     6001 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6061 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6181 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6241 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6301 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6361 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6421 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6481 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6541 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6661 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6721 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6781 ctgattctgt ggataaccgt attaccgcca tgcat
//
