LOCUS       sequence-333421-        6752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses APEX2 linked to tau for labeling microtubules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6752)
  TITLE     Expression plasmids combining APEX2 with various fluorophores and 
            targeting sequences
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6752)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV_hsyn_FLEX_tau-APEX2
FEATURES             Location/Qualifiers
     source          1..6752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   52..181
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        226..673
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    716..749
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    794..827
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(2027..2779)
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="MVGKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSA
                     GTFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVV
                     AVEVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGG
                     HTIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLV
                     DKYAADEDAFFADYAEAHQKLSELGFADA"
     protein_bind    complement(2785..2818)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2869..2902)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    2930..3518
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2983..3003)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3401..3412)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3550..4026
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3749..3768)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4066..4206
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4066..4195
                     /label=AAV2 ITR
     rep_origin      4281..4736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4368..4387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4578..4599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4753..4772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4872..4894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4932..4950)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5018..5122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5123..5191,5192..5983)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5341..5360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6154..6742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6643..6662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
       61 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
      121 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      181 tgcggccgca cgcgtgtgtc tagactgcag agggccctgc gtatgagtgc aagtgggttt
      241 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      301 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      361 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      421 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      481 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      541 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      601 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      661 cctgagagcg cagtcgagaa ggtaccggat ccaggccacc atgtctagag tcgacataac
      721 ttcgtataat gtatgctata cgaagttatg cagaatggta gcttaggctg ctattagcaa
      781 tatgaaacct cttataactt cgtataaagt atcctatacg aagttatgtt aattaatatc
      841 acaaaccctg cttggccagg gaggcagaca cctcgtcagc tagcgtggcg agctggggcg
      901 agtctaccat gtcgatgctg ccggtggagg agacattgct gagatgccgt ggagacgtgt
      961 ccccagacac cactggcgac ttgtacacga tctccgcccc gtggtctgtc ttggctttgg
     1021 cgttctcgcg gaaggtcagc ttgtgggttt caatcttttt atttcctccg ccagggacgt
     1081 gggtgatatt gtccagggac ccaatcttcg actggactct gtccttgaag tcaagcttct
     1141 cagattttac ttccacctgg ccacctcctg gtttatgatg gatgttgcct aatgagccac
     1201 acttggaggt caccttgctc aggtcaactg gtttgtagac tatttgcaca ctgccgcctc
     1261 ccgggacgtg tttgatatta tcctttgagc cacacttgga ctggacgttg ctaagatcca
     1321 gcttcttatt aattatctgc accttcccgc ctcccggctg gtgcttcagg ttctcagtgg
     1381 agccgatctt ggacttgaca ttcttcaggt ctggcatggg cacgggggct gtctgcaggc
     1441 ggctcttggc ggaagacggc gacttgggtg gagtacggac cactgccacc ttcttgggct
     1501 cccgggtggg tggggttgga agggacgggg tgcgggagcg gctgccggga gtgcctgggg
     1561 agccggggct gctgtagccg ctgcgatccc ctgattttgg aggttcacca gagctgggtg
     1621 gtgtctttgg agcgggcggg gtttttgctg gaatcctggt ggcgttggcc tggcccttct
     1681 ggcctggagg ggctgctccc cgcggtgtgg cgatcttcgt tttaccatca gcccccttgg
     1741 cttttttgtc atcgcttcca gtcccgtctt tgcttttact gaccatgcga gcttgggtca
     1801 cgtgaccagc agcttcgtct tccaggctgg gggtgtctcc aatgcctgct tcttcagctt
     1861 tcaggccagc gtccgtgtca ccctcttggt cttggtgcat ggtgtagccc ccctgatctt
     1921 tcctgtcccc caacccgtac gtcccagcgt gatcttccat cacttcgaac tcctggcggg
     1981 gctcagcgcc cgagcccgag gtcgagcccg agcccttggc gcgcccggca tcagcaaacc
     2041 caagctcgga aagcttttgg tgagcctcag cgtaatcagc aaagaaggca tcttcgtccg
     2101 ctgcatattt gtcaacgaga gggcggaata cagggtcaga caaaagagcc ttgtcagaag
     2161 gtagctgaag gagaccttcc ttctcaccac tcaacaactc cgtgaagtat gagttgtcga
     2221 aaataagagg attagaggtc cagggaccct caaatccaga acgctccttg tgtgcagctc
     2281 caatagtgtg acccccagat agagcaacga tatcttggtc agtaagcccc atagctttgc
     2341 caaacacatc tctcaaatgg tcagaaccct tagtgggatc gggcaagcga ccctctggtg
     2401 gtggctcagg cttgtcctct cttccagggt ggaatggaac cttaggtcca cccgtgacct
     2461 caacggcaac aacgccagcc aactggtaga aatcggcgta gctcaaaata gggaactccg
     2521 ccttgagtgg ctccaaaagc ctaacagcga tgtcaagacc gttgttagcg ctgtgagcca
     2581 gttcggcagg gtgcttgatg gttccgaagg gtccaccggt cttcgtgccc ttgtcaaagg
     2641 ttccagcaga gtggaatgcc aaacggagca ttagaggagc gcatctcttc tcagcgatga
     2701 agcctctgag cttcttcttc gccttctcaa cggcgtcctg gtaatcagca ctcacagttg
     2761 ggtaagactt tcctaccatg gcccataact tcgtatagca tacattatac gaagttattt
     2821 gccttaaccc agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat
     2881 aggatacttt atacgaagtt attgagaatt cgatatcaag cttatcgata atcaacctct
     2941 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3001 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3061 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3121 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3181 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3241 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3301 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     3361 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3421 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3481 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3541 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3601 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3661 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3721 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     3781 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     3841 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     3901 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3961 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     4021 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4081 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4141 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4201 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4261 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4321 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4381 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4441 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4501 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4561 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4621 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4681 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4741 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4801 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4861 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4921 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4981 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5041 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5101 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5161 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5221 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5281 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5341 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5401 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5461 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5521 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5581 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5641 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5701 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5761 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5821 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5881 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5941 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6001 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6061 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6301 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6721 aggggggcgg agcctatgga aaaacgccag ca
//
