LOCUS       sequence-412569-        7152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synaptic activity can be imaged with Synaptophysin GCaMP, while 
            simultaneously, a red fluorescence structural marker can be employed
            to provide a visual reference for the cellular architecture. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7152)
  TITLE     Prigge Lab AAV vectors
REFERENCE   2  (bases 1 to 7152)
  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"
COMMENT     Sequence Label: pAAV_CbH_SIO_SyGCaMP8s_p2a_mRuby
FEATURES             Location/Qualifiers
     source          1..7152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        145..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          655..882
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     protein_bind    complement(894..927)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(940..1647)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     CDS             complement(1648..1704)
                     /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"
     primer_bind     2270..2289
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2529..2550
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2656..2677)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3885..3894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3903..3936
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4007..4027)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4415..4555
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4415..4544
                     /label=AAV2 ITR
     rep_origin      4630..5085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4717..4736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4927..4948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5102..5121)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5221..5243
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5281..5299)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5367..5471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5472..5540,5541..6332)
                     /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(5690..5709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6503..7091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6992..7011
                     /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 gcgtcgttac ataacttacg gtaaatggcc cgcctggctg
      181 accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg
      241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc
      361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      421 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      481 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      541 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      601 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      661 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
      721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      781 tgtaattagc tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg
      841 tttaattacc tggagcacct gcctgaaatc actttttttc aggttgggga tcctaccgtt
      901 cgtatagcat acattatacg aagttatgat ttctaattac ttgtacagct cgtccatgcc
      961 accaccaagg ttgctgtatt tggcaactgc cacttctctt tgcactacgt aggtttcatt
     1021 gtcactctcc tcgatccttt ccaggcggtg atcaacggca tggacaccgg gcatcttgat
     1081 gttcccgacg gtcttttttg acctgtaagt tgtcacgaag ttgcagtgca gatggccacc
     1141 accatcaact ttcagtgcga tgtcagtgta tcctctcaga ccaccatctg ctggatacat
     1201 catctctgta ttaggctccc aacccttggt cttcttctgc atcacgggac cattggaggg
     1261 aaagtttacc cctctgacct tgacgttgta gacgagctcg ccatcctcaa ggctggtgtc
     1321 ctgcgtgacg gtgacgactc caccatcttc gtatctcgta actctttccc aagtaaaacc
     1381 ctcaggaaag gactgtttaa agaaatcagg gatgtcggcc gggtacttga taaaggtacg
     1441 gctgccatac atgaacgacg tggcaagaat gtcaaaggca aatggcaggg gtcctccctc
     1501 gatgactttg atcctcatgg tttgcactcc ctcgtacggt ctgccttctc cttcacctgt
     1561 gcatttgaat tggtggccgt tgaccgaacc ttccatgacc accttcatac gcatattttc
     1621 cttgatcagc tcttcgccct tagacacagg accggggttt tcttccacgt ctcctgcttg
     1681 ctttaacaga gagaagttcg tggctccgga gcccttcgct gtcatcattt gtacaaactc
     1741 ttcgtagttt acctgaccat ccccatcgat gtctgcttcc ctgatcattt catcaacctc
     1801 ttcatctgtt aacttctctc caaggtttgt catcacgtgg cgaagctctg ctgcactgat
     1861 gtagccattg ccatccttat caaacacacc gaacgcttct ctaatttctt cttccgtgtc
     1921 cctgtatttc atttttcttg ccatcattgt caggaactca gggaagtcga ttgtgccgtc
     1981 accgtcggca tctacttcat tgatcatgtc ctgcagctct gcttctgtgg ggttgtgtcc
     2041 gagagaccgc atcaccgtcc ccagctcctt ggttgttatt gtcccatccc cgtccttgtc
     2101 aaatagggag aaagcctctt tgtattctgc gatctgctct tcagtcagtt ggtccggcag
     2161 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     2221 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     2281 gccgtcgtcc ttgaagaaga tggtgcgctc ctggatgtag ccttcgggca tggcggactt
     2341 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     2401 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     2461 cagggtcagc ttgccgtagg tggcatcgcc ctcaccctcg ccggacacgc tgaacttgtg
     2521 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     2581 gcccttgctc accatgctcc ctccggtacc gcccttgtac agctcgtcca tgccgagagt
     2641 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     2701 tttcgaaagt ttggactcca cgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     2761 gccgatgggg gtgttctgct ggtagtggta ggcgagctgc acgccgccgt cctcgatgtt
     2821 gtggcggatg tggaagttcg ccttgatgcc gttcttctgc ttgtcggcct tgatatagac
     2881 gttgatcttc aggcccatca gcatggcgat gatcttcacg gcggtggcca cctccttgaa
     2941 ggtcttcttg cgacgcgtta ccggtggagc ctgattggag aaggaggtgg gcgcaccctg
     3001 ttggccatag ccttgctgcc catagtcgcc ctgaggcccg tagccaccgc caccgctggc
     3061 tggctgcccg taatcgggtt gataaccacc ctgaggcccg taggagtcct ggggcccata
     3121 gcctccgggg ccctgcccgt agcccgcatc gccgtaggca tcgccaggtg ctggttgctt
     3181 ttccggggcg cctggaggtg cgcgcatgaa tggggctgcc cagcctgtct ccttgaacac
     3241 gaaccataag ttgccaaccc agagcaccag gttcaggaag ccaaacacca ctgtggtgtt
     3301 gagtcctgaa gtcacagggt ccctcagttc cttgcatgtg ttccctgtct ggcggcacat
     3361 gggcatctcc ttgataatgt tctctgggtc cgtggccatc ttcacatcgg acaggccttt
     3421 ggcccaggct gatgaactaa ctagccacat gaaagcgaac acggctgtag ccagaaagtc
     3481 catcataggc cctttgttgt tctctcggta cttgttctgc aggaagatgt aggtggccag
     3541 ggcccccatg gagtagagga aggcaaacac agccacggtg acaaagaact cagccgacga
     3601 ggagtagtcc cccaccagga agatcttggt agtgccccct ttgacgcagg agggtgcatc
     3661 aaagtacact tggtgcagcc tgaaggggta ctcgaactca acttcgatgt tgagggcact
     3721 ctccgtcttg ttggcacact ccacgctcag ccgaagctcc ccggtgtagc tgccacacgt
     3781 agcaaaggcg aagatggcaa agacccactg cagcaccttc acgaagccaa ggggctcctt
     3841 gaccacccgg aactgacccc cagccaccag ctgattcacc acgtccatgg tggcggctag
     3901 cctaccgttc gtataatgta tgctatacga agttatggaa ttccgctcga gataatcaac
     3961 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     4021 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     4081 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     4141 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4201 ttatttgtga aatttgtgat gctattgctt tatttgtaac catctagctt tatttgtgaa
     4261 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4321 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     4381 ggccgcggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4441 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4501 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4561 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4621 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4681 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     4741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4921 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4981 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     5041 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5101 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5161 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5221 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5281 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5341 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5401 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5461 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5521 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5581 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5641 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5701 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5761 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5821 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5881 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5941 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6001 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6061 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6121 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6181 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6241 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6301 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6361 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6421 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6481 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6541 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6601 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6661 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6721 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6781 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6841 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6901 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6961 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7021 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7081 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7141 ttgctcacat gt
//
