LOCUS       sequence-225121-        7482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid name in publication: pAAV-DIO-SV-HRP. Peroxidase reporter 
            for multiplexed electron microscopy labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7482)
  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 7482)
  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-SYP-HRP
FEATURES             Location/Qualifiers
     source          1..7482
                     /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)."
     regulatory      3450..3459
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3467..3500
                     /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    3551..3584
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3609..4197
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3662..3682)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4080..4091)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4229..4705
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4428..4447)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4745..4885
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4745..4874
                     /label=AAV2 ITR
     rep_origin      4960..5415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5047..5066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5257..5278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5432..5451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5551..5573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5611..5629)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5697..5801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5802..5870,5871..6662)
                     /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(6020..6039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6833..7421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7322..7341
                     /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 tgttacatct gattggagaa ggaggtgggc gcaccctgtt ggccgtagcc
     1621 ttgctgccca tagtcgccct gaggcccgta gccaccgcca ccgccgctgg ctggctgccc
     1681 gtaatcgggt tgataaccac cctgaggccc gtaggagtcc tggggcccat agcctccggg
     1741 gccctgccca tagcccgcat cgccgtaggc atcgccagga gctggttgct tttctggggc
     1801 gcctggaggt gcgcgcatga atggggcggc ccagcctgtc tccttgaaca cgaaccatag
     1861 gttgccaacc cagagcacca ggttcaggaa gccaaacacc accgaggtgt tgagtcctga
     1921 agtcacaggg tccctcagtt ccttgcatgt gattcgagct ccaccggtag agttgctgtt
     1981 gaccactctg cagttcagac gaatctggcc ttgggtaccc gtcagagggg taatgttacc
     2041 catacggtcc atggcttcca cgaaggcgtt aaagaaggtt tgagtagagt tagcaaaact
     2101 tctcaccagt gggatggtgt cagtggcgtt tggactgcta aacagttctt gatcactctg
     2161 tatcaggcct ttctgctcct ctagattcac atagtacttg ttatcgaaga tggttggggt
     2221 ccgcagatca aagtccacta gtgcactgag gttgccattc agtgggcaca agcctctcag
     2281 tgtctggaga tacgtagtgt tcagcgtggg gtcaggtaac ccagtgttgc tgaaattgta
     2341 gagcctatcc atgatgaacc tacactggtt ctttccaaat gtgtgtcctc cggacagagc
     2401 cacaaggtca ctcgagcgat tcagacccac gtttctaaag ctatccttca gctggggcag
     2461 ggtgaagaat ggagcaggca agttggcgtt ggccagatct aggaatgcct gtagggagtc
     2521 acgtcgaccg agcggcactc tccaggacgg tccgcctgca agagtcacgc tctgttgcgc
     2581 agctatagtc agcaggtctg cacaactgac tgttcgtggg catgctgact caacggcagc
     2641 cttcatgcga tcgatcactg gaaagcccct ggcgctgtta gcgttcccga atgcatcctt
     2701 ttcagtgcgg aaactggtgg tgttgtccag taatatgcta gcgtcgcaac cattcacgaa
     2761 gcagtcatgg aagtgcagac gtaatattga agcagcgatc ctgggatcgg atctgagctc
     2821 gttgacgatt gtgtcgcgaa cgatgttgga cacgttggga cagctattgt cgtagaatgt
     2881 aggggttaac tgggtggcta gatccatatc tgtctggcgg cacataggca tctccttgat
     2941 aatgttctct gggtcagtgg ccatcttcac atcggacagg cctttggccc aggcggatga
     3001 gctaactagc cacatgaaag cgaacactgc tgtggccagg aagtccatca ttggcccttt
     3061 gttgttctct cggtacttgt tctgcaggaa gatgtaggtg gccagggccc ccatggagta
     3121 gaggaaggca aacacagcca cggtgacaaa gaattcagcc gaggaggagt agtcaccaac
     3181 taggaagatc ttggtagtgc cccctttaac gcaggagggt gcatcaaagt acacttggtg
     3241 cagcctgaat gggtactcaa attcgacttc gatgttgagg gcactctccg tcttgttggc
     3301 acactccacg ctcagccgaa gctctccggt gtagctgccg cacgtagcaa aggcgaagat
     3361 ggcaaagacc cactgcagca ccttcacgaa gccaaggggc tccttgacca cccggaactg
     3421 acccccagcc accagctgat tcaccacgtc catggtggcc atgagcataa cttcgtataa
     3481 agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat
     3541 ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc
     3601 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3661 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3721 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3781 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3841 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3901 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3961 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     4021 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     4081 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     4141 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     4201 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     4261 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     4321 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     4381 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     4441 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4501 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4561 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4621 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4681 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4741 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4801 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4861 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4921 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4981 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     5041 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5101 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5161 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5221 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5281 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     5341 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5401 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5461 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5521 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5581 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5641 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5701 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5761 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5821 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5881 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5941 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6001 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6061 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6121 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6181 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6241 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6301 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6361 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6421 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6481 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6541 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6601 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6661 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6721 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6781 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6841 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6901 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6961 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7021 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7081 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7141 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7201 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7261 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7321 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7381 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7441 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
