LOCUS       sequence-430558-        7275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid to express H2B-BFP and codon-optimized Rabies G
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7275)
  AUTHORS   Patino M, Lagos WN, Patne NS, Miyazaki PA, Bhamidipati SK, Collman 
            F, Callaway EM
  TITLE     Postsynaptic cell type and synaptic distance do not determine 
            efficiency of monosynaptic rabies virus spread measured at synaptic 
            resolution.
  JOURNAL   Elife. 2023 Dec 14;12:e89297. doi: 10.7554/eLife.89297.
  PUBMED    38096019
REFERENCE   2  (bases 1 to 7275)
  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: "Elife."; 
            date: "2023-12-14"; volume: "12:e89297. doi"; pages: " 
            10.7554/eLife.89297"
COMMENT     Sequence Label: pAAV-hSyn-FLEX-H2BmTagBFP2-oG
FEATURES             Location/Qualifiers
     source          1..7275
                     /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        158..605
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      612..621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    627..660
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    711..744
                     /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(2326..2382)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(2398..2424)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2428..2454)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2458..2484)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3192..3201
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3214..3591)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     protein_bind    complement(3597..3630)
                     /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(3681..3714)
                     /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    3716..4304
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3769..3789)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4187..4198)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4312..4536
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4538..4678
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4538..4667
                     /label=AAV2 ITR
     rep_origin      4753..5208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4840..4859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5050..5071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5225..5244)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5344..5366
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5404..5422)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5490..5594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5595..5663,5664..6455)
                     /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(5813..5832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6626..7214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7115..7134
                     /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 gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagtcagc cgccaccatg gttaatataa cttcgtataa tgtatgctat acgaagttat
      661 gcagaatggt agctggattg tagctgctat tagcaatatg aaacctctta ataacttcgt
      721 ataaagtatc ctatacgaag ttatggtacc ttagagccgt gtctcgcccc cagatttatg
      781 actttcccaa ctgctgatga ttttaccgct ctggggtgtg acagacactt ctctaccggt
      841 ccccctcaga ttgtgctgag ttggctcgga gcggttcact cgtcgccagc aggtcatcag
      901 gaaaatgatc agcatgagag cagtcagtgc tccagcggag agcagcacgt acttgcccca
      961 attaggcagt ccgagatcga cgccactgat ccgttcgtgc acgtcgggca ggtggacctc
     1021 cacaaaatct tcggcctcgt cgccgttttt gaacactgtg cttgggtctg cgagaggatg
     1081 catcagggga atgacgcttg acacgagcag ttccatgtgc tgctggagca gactagactg
     1141 catctcgggg atcagcacgt taccatctgg ccccagaatg ataccattaa agaagacccc
     1201 gttcacatgt gggtgacatc gacccccgac tcgcaggcat cccttactag gaatgatttc
     1261 attccaggtt ctcacagatt tataatgggc gtctgcctcc atcagtgtct tgttaaagat
     1321 ggtgtatgct tttccgaagc cagggacgag cttcctcagg tggctcagcc tcctgaagct
     1381 cacgctcttg gtagtcataa ttgactccag agcatcgaga cattcctccc ttttcttcac
     1441 gagttcctcg accaccagat gttcgatctc atcgcttcgg aagtcgtgca gattcacgag
     1501 ctgcccaggg gggcaccact tagtctcgtt agatgtctgc atagcgaccc aggttccgtc
     1561 catcagccgg agtcccagca cgccacagag tttcagcttg caggcacctt tgaggctctt
     1621 atacagccct ctttcatcca caaatccaca agtctcgctg ccttttgagg ccctcttgcc
     1681 cctgctgttg gtgaagatgt cgcaggacat ccccagcctt ggattctcag gcatccagat
     1741 tgtataatca tggttggtgg aacagtaagt ggagctgacg gccactccgg agcagtttcc
     1801 gccgggaaac actggactgt ggagagatcg gtcgtagggg tccaggtcgg cgacagatgg
     1861 ggaaatgatg accaggctct cttttgtggt cttcacagtg cgcagccaat gatagtctgg
     1921 gtaaggattg tgcaggcttt cctcatagcg ggggtcgcca gccatcttcc agttgtaagc
     1981 tgcccgacag gcgtctggtg ttggccgaaa gtgttttctc ttgaaagttg tggtgacata
     2041 tcccacaaaa tttgtgtagg tttctgcctc ggtgaccaca ccagtgcatg tgaacccgtt
     2101 catcttaata gcagagatat agcccacttt cagctccatg taggagaatc cgctcaggtt
     2161 ggtacagcct tcgtcctcga ccacgagatt gttagggcat gacagatggt gaatatcgat
     2221 aggactccag gggcccagct tgtcggggat agtgtagata gggaatttac caaagcagag
     2281 tgggaagacg agcagaggga cgaagaggag agcctgtggg accatgggtc cagggttctc
     2341 ctccacgtct ccagcctgct tcagcaggct gaagttagta gctccggatc ctccgccagc
     2401 gtaatctgga acgtcatatg gatatcctgc atagtccggg acgtcatagg gatagcccgc
     2461 atagtcagga acatcgtatg ggtagttgag cttatgaccc aacttactag gcaaatcaca
     2521 gtagcgggcg acggcgactt cgtgctgctc gacgtaggtt tcgttgttag cttccttaat
     2581 gcgctccagc ctatagtcca cgtagtacac accgggcatc ttcaggttct tcgctggctt
     2641 cttactacgg tacgtagtct tggcattggc tataaggtgc gaaccgccca ccagcttcag
     2701 agccatatcg ttgcgacctt caagaccgcc atcggcgggg tacagcgtct ccgtaaaggc
     2761 ctcccatccg agcgtctttt tctgcataac cgggccgttc gaggtaaagt tcacgccgcg
     2821 tatcttcacg ttgtaaatca agcaaccatc ctgcaggcta gtatcttggg tggcagtcag
     2881 aactccgccg tcctcatagg tagtaacgcg ttcccatgtg aagccttcag ggaaggattg
     2941 cttgaagaaa tctggtattc cctgagtatg attgatgaat gtcttgctgc catagaggaa
     3001 actggtggcc aggatatcga atgcaaatgg cagtggacct ccctcgacaa ctttaatacg
     3061 catggtctgt gtgccctcgt agggctttcc ctcaccctcg ctggtgcact taaagtggtg
     3121 gttgtcgacg gttccctcca tatacagctt catgtgcata ttctctttga tcagctcctc
     3181 tcccttcgaa accatggtgg cgaccggtgg atccttagcg ctggtgtact tggtgatggc
     3241 cttagtaccc tcggacacgg cgtgcttggc caactcccca ggcagcagca ggcgcacggc
     3301 cgtctggatc tccctggagg tgatggtcga gcgcttgttg taatgcgcca ggcgggaagc
     3361 ctcacctgcg atgcgctcga aaatgtcgtt cacaaacgaa ttcatgatgc ccatggcctt
     3421 ggacgaaatg ccggtgtcag ggtggacctg cttcagaacc ttgtacacat agatggaata
     3481 gctctccttg cggctgcgct tgcgcttctt gccgcctttc ttctgcgcct tagtcaccgc
     3541 cttcttggag ccctttttcg gggcgggagc agacttcgct ggctctggca tacctaataa
     3601 cttcgtatag catacattat acgaagttat ttgccttaac ccagaaatta tcactgttat
     3661 tctttagaat gatgcaaaga ataacttcgt ataggatact ttatacgaag ttatgaatca
     3721 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3781 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3841 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3901 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3961 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4021 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4081 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     4141 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4201 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4261 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggat cctgtgcctt
     4321 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4381 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4441 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4501 atagcaggca tgctggggat gcggtgggct ctatggcagg aacccctagt gatggagttg
     4561 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4621 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     4681 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     4741 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4801 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     4861 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4921 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4981 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5041 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     5101 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     5161 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5221 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5281 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5341 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5401 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5461 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5521 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5581 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5641 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5701 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5761 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5821 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5881 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5941 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6001 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6061 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6121 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6181 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6241 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6301 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6361 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6421 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6481 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6541 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6601 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6661 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6721 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6781 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6841 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6901 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6961 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7021 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7081 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7141 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7201 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7261 cttttgctca catgt
//
