LOCUS       sequence-328048-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HA-BACE1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  AUTHORS   Prabhu Y, Burgos PV, Schindler C, Farias GG, Magadan JG, Bonifacino 
            JS
  TITLE     Adaptor protein 2-mediated endocytosis of the beta-secretase BACE1 
            is dispensable for amyloid precursor protein processing.
  JOURNAL   Mol Biol Cell. 2012 Jun;23(12):2339-51. doi: 
            10.1091/mbc.E11-11-0944. Epub 2012 May 2.
  PUBMED    22553349
REFERENCE   2  (bases 1 to 6245)
  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: 
            "10.1091/mbc.E11-11-0944"; journalName: "Mol Biol Cell"; date: 
            "2012-06"; volume: "23"; issue: "12"; pages: "2339-51"
COMMENT     Sequence Label: GFP-HA-BACE1
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             700..726
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2125..2134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2131..2133,2134..2136,2137..2850)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2176..2197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2437..2456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2784..2805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2973..3094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3010..3029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3064..3083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3101..3556)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3238..3259)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3450..3469
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3583..3687
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3689..4046
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3689..3880
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3712..3732)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3897..4032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4081..4875
                     /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             4081..4875
                     /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(4135..4154)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4745..4764
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5063..5082)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5107..5154
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5483..6071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5972..5991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagtt caccatggcc caagccctgc cctggctcct gctgtggatg
      601 ggcgcgggag tgctgcctgc ccacggcacc cagcacggca tccggctgcc cctgcgcagc
      661 ggcctggggg gcgcccccct ggggctgcgg ctgccccggt acccttacga tgtaccagat
      721 tacgctgaga ccgacgaaga gcccgaggag cccggccgga ggggcagctt tgtggagatg
      781 gtggacaacc tgaggggcaa gtcggggcag ggctactacg tggagatgac cgtgggcagc
      841 cccccgcaga cgctcaacat cctggtggat acaggcagca gtaactttgc agtgggtgct
      901 gccccccacc ccttcctgca tcgctactac cagaggcagc tgtccagcac ataccgggac
      961 ctccggaagg gtgtgtatgt gccctacacc cagggcaagt gggaagggga gctgggcacc
     1021 gacctggtaa gcatccccca tggccccaac gtcactgtgc gtgccaacat tgctgccatc
     1081 actgaatcag acaagttctt catcaacggc tccaactggg aaggcatcct ggggctggcc
     1141 tatgctgaga ttgccaggcc tgacgactcc ctggagcctt tctttgactc tctggtaaag
     1201 cagacccacg ttcccaacct cttctccctg cagctttgtg gtgctggctt ccccctcaac
     1261 cagtctgaag tgctggcctc tgtcggaggg agcatgatca ttggaggtat cgaccactcg
     1321 ctgtacacag gcagtctctg gtatacaccc atccggcggg agtggtatta tgaggtgatc
     1381 attgtgcggg tggagatcaa tggacaggat ctgaaaatgg actgcaagga gtacaactat
     1441 gacaagagca ttgtggacag tggcaccacc aaccttcgtt tgcccaagaa agtgtttgaa
     1501 gctgcagtca aatccatcaa ggcagcctcc tccacggaga agttccctga tggtttctgg
     1561 ctaggagagc agctggtgtg ctggcaagca ggcaccaccc cttggaacat tttcccagtc
     1621 atctcactct acctaatggg tgaggttacc aaccagtcct tccgcatcac catccttccg
     1681 cagcaatacc tgcggccagt ggaagatgtg gccacgtccc aagacgactg ttacaagttt
     1741 gccatctcac agtcatccac gggcactgtt atgggagctg ttatcatgga gggcttctac
     1801 gttgtctttg atcgggcccg aaaacgaatt ggctttgctg tcagcgcttg ccatgtgcac
     1861 gatgagttca ggacggcagc ggtggaaggc ccttttgtca ccttggacat ggaagactgt
     1921 ggctacaaca ttccacagac agatgagtca accctcatga ccatagccta tgtcatggct
     1981 gccatctgcg ccctcttcat gctgccactc tgcctcatgg tgtgtcagtg gcgctgcctc
     2041 cgctgcctgc gccagcagca tgatgacttt gctgatgaca tctccctgct gaagacggta
     2101 ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     2161 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2221 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     2281 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2341 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2401 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2461 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2521 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2581 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2641 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2701 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     2761 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2821 actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc
     2881 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2941 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3001 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3061 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     3121 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3181 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3241 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3301 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3361 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3421 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3481 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3541 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3661 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3721 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3841 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3901 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3961 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4021 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4081 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4141 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4201 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4261 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4321 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4381 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4441 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4501 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4561 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4621 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4681 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4741 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4801 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4861 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4921 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4981 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5041 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5101 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5161 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5221 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5281 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5341 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5401 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5461 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5521 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5581 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5641 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5701 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5761 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5821 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5881 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5941 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6001 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6061 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6121 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6181 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     6241 ttccg
//
