LOCUS       sequence-386452-        7042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Bace1 with an HA tag at N-terminal for single molecule 
            tracking and other measusers in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7042)
  AUTHORS   Capitini C, Bigi A, Parenti N, Emanuele M, Bianchi N, Cascella R, 
            Cecchi C, Maggi L, Annunziato F, Pavone FS, Calamai M
  TITLE     APP and Bace1: Differential effect of cholesterol enrichment on 
            processing and plasma membrane mobility.
  JOURNAL   iScience. 2023 Apr 11;26(5):106611. doi: 10.1016/j.isci.2023.106611.
            eCollection 2023 May 19.
  PUBMED    37128606
REFERENCE   2  (bases 1 to 7042)
  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: "iScience.";
            date: "2023-04-11"; volume: "26(5):106611. doi"; pages: " 
            10.1016/j.isci.2023.106611. eCollection 2023 May 19"
COMMENT     Sequence Label: HA-BACE1-mBFP
FEATURES             Location/Qualifiers
     source          1..7042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1098..1124
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2592..3293
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(3299..3315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3299..3315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3326..3948
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3525..3544)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4097..4685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4177..4196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5014..5061
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5086..5105
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5293..6087)
                     /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             complement(5293..6087)
                     /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(5404..5423)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6014..6033
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6122..6479)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6136..6271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6288..6479
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6436..6456
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6481..6585)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6612..7040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6699..6718)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6909..6930
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggcccaagcc ctgccctggc tcctgctgtg gatgggcgcg ggagtgctgc
     1081 ctgcccacgg cgctagctat ccgtacgacg taccagacta cgcaggcgcg cctacccagc
     1141 acggcatccg gctgcccctg cgcagcggcc tggggggcgc ccccctgggg ctgcggctgc
     1201 cccgggagac cgacgaagag cccgaggagc ccggccggag gggcagcttt gtggagatgg
     1261 tggacaacct gaggggcaag tcggggcagg gctactacgt ggagatgacc gtgggcagcc
     1321 ccccgcagac gctcaacatc ctggtggata caggcagcag taactttgca gtgggtgctg
     1381 ccccccaccc cttcctgcat cgctactacc agaggcagct gtccagcaca taccgggacc
     1441 tccggaaggg tgtgtatgtg ccctacaccc agggcaagtg ggaaggggag ctgggcaccg
     1501 acctggtaag catcccccat ggccccaacg tcactgtgcg tgccaacatt gctgccatca
     1561 ctgaatcaga caagttcttc atcaacggct ccaactggga aggcatcctg gggctggcct
     1621 atgctgagat tgccaggcct gacgactccc tggagccttt ctttgactct ctggtaaagc
     1681 agacccacgt tcccaacctc ttctccctgc agctttgtgg tgctggcttc cccctcaacc
     1741 agtctgaagt gctggcctct gtcggaggga gcatgatcat tggaggtatc gaccactcgc
     1801 tgtacacagg cagtctctgg tatacaccca tccggcggga gtggtattat gaggtgatca
     1861 ttgtgcgggt ggagatcaat ggacaggatc tgaaaatgga ctgcaaggag tacaactatg
     1921 acaagagcat tgtggacagt ggcaccacca accttcgttt gcccaagaaa gtgtttgaag
     1981 ctgcagtcaa atccatcaag gcagcctcct ccacggagaa gttccctgat ggtttctggc
     2041 taggagagca gctggtgtgc tggcaagcag gcaccacccc ttggaacatt ttcccagtca
     2101 tctcactcta cctaatgggt gaggttacca accagtcctt ccgcatcacc atccttccgc
     2161 agcaatacct gcggccagtg gaagatgtgg ccacgtccca agacgactgt tacaagtttg
     2221 ccatctcaca gtcatccacg ggcactgtta tgggagctgt tatcatggag ggcttctacg
     2281 ttgtctttga tcgggcccga aaacgaattg gctttgctgt cagcgcttgc catgtgcacg
     2341 atgagttcag gacggcagcg gtggaaggcc cttttgtcac cttggacatg gaagactgtg
     2401 gctacaacat tccacagaca gatgagtcaa ccctcatgac catagcctat gtcatggctg
     2461 ccatctgcgc cctcttcatg ctgccactct gcctcatggt gtgtcagtgg cgctgcctcc
     2521 gctgcctgcg ccagcagcat gatgactttg ctgatgacat ctccctgctg aagacgcgta
     2581 cgcggccgcg aatgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
     2641 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     2701 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     2761 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
     2821 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     2881 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     2941 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
     3001 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
     3061 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac gccaagacca
     3121 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact
     3181 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg
     3241 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat taaccgcggt
     3301 catagctgtt tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct
     3361 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3421 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3481 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3541 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3601 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3661 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3721 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3781 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
     3841 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
     3901 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
     3961 agcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4021 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4081 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4141 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4201 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4261 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4321 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4381 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4441 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4501 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4561 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4621 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4681 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4741 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4801 aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc
     4861 ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg
     4921 gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag
     4981 agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt
     5041 tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg
     5101 tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga
     5161 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     5221 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc
     5281 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     5341 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     5401 agcaatatca cgggtagcca acgctatgtc ctgatagcga tccgccacac ccagccggcc
     5461 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     5521 gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag
     5581 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     5641 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     5701 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     5761 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     5821 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     5881 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt
     5941 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     6001 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     6061 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat
     6121 ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     6181 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     6241 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     6301 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     6361 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     6421 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg
     6481 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6541 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6601 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6961 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7021 acaaaaattt aacgcgaatt tt
//
