LOCUS       sequence-225611-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Peroxidase reporter for multiplexed electron microscopy labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  AUTHORS   Zhang Q, Lee WA, Paul DL, Ginty DD
  TITLE     Multiplexed peroxidase-based electron microscopy labeling enables 
            simultaneous visualization of multiple cell types.
  JOURNAL   Nat Neurosci. 2019 Mar 18. pii: 10.1038/s41593-019-0358-7. doi: 
            10.1038/s41593-019-0358-7.
  PUBMED    30886406
REFERENCE   2  (bases 1 to 6357)
  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 
            Neurosci. 2019 Mar 18. pii: 10.1038/s41593-019-0358-7. doi: 
            10.1038/s41593-019-0358-7."
COMMENT     Sequence Label: pAAV-DIO-dAPEX2
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1580..2329)
                     /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="MGKSYPTVSADYQKAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPEVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
     regulatory      2326..2335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2342..2375
                     /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)."
     protein_bind    2426..2459
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2484..3072
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2537..2557)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2955..2966)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3104..3580
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3303..3322)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3620..3760
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3620..3749
                     /label=AAV2 ITR
     rep_origin      3835..4290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3922..3941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4132..4153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4307..4326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4426..4448
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4486..4504)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4572..4676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4677..4745,4746..5537)
                     /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(4895..4914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5708..6296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6197..6216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttact tgtacactag gcatcagcaa acccaagctc ggaaagcttt tggtgagcct
     1621 cagcgtaatc agcaaagaag gcatcttcgt ccgctgcata tttatcaacg agagggcgga
     1681 atacagggtc agacaaaaga gccttgtcag aaggtagctg aaggagacct tccttctcac
     1741 cactcaacaa ctccgtgaag tatgagttgt cgaaaataag aggattagag gtccagggac
     1801 cctcaaatcc agaacgctcc ttgtgtgcag ctccaatagt gtgaccccca gatagagcaa
     1861 cgatatcttg gtcagtaagc cccatagctt tgccaaacac atctctcaaa tggtcagaac
     1921 ccttagtggg atcgggcaag cgaccctctg gtggtggctc aggcttgtcc tctcttccag
     1981 ggtggaatgg aacttcaggt ccacccgtga cctcaacggc aacaacgcca gccaactggt
     2041 agaaatcggc gtagctcaaa atagggaact ccgccttgag tggctccaaa agcctaacag
     2101 cgatgtcaag accgttgtta gcgctgtgag ccagttcggc agggtgcttg atggttccga
     2161 agggtccacc ggtcttcgtg cccttgtcaa aggttccagc agagtggaat gccaaacgga
     2221 gcattagagg agcgcatctc ttctcagcga tgaagcctct gagcttcttc ttcgccttct
     2281 caacggcctt ctggtaatca gcactcacag ttgggtaaga ctttcccatg gtggcgctag
     2341 cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact
     2401 gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg
     2461 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2521 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2581 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2641 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2701 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2761 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2821 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2881 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     2941 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3001 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3061 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     3121 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     3181 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     3241 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     3301 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     3361 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     3421 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     3481 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     3541 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     3601 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3661 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3721 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3781 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3841 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3901 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3961 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4021 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4141 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     4201 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4261 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4321 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4381 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4441 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4501 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4561 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4621 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4681 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4741 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4801 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4861 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4921 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4981 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5041 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5101 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5161 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5221 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5281 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5341 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5401 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5461 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5521 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5581 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5641 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5701 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5761 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5821 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5881 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5941 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6001 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6061 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6121 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6181 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6241 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6301 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
