LOCUS       sequence-371325-        7768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iDREADD fused to mCherry flanked by DIO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7768)
  AUTHORS   Nasrallah K, Frechou MA, Yoon YJ, Persaud S, Goncalves JT, Castillo 
            PE
  TITLE     Seizure-induced strengthening of a recurrent excitatory circuit in 
            the dentate gyrus is proconvulsant.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Aug 9;119(32):e2201151119. doi: 
            10.1073/pnas.2201151119. Epub 2022 Aug 5.
  PUBMED    35930664
REFERENCE   2  (bases 1 to 7768)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-08-9"; volume: "119(32):e2201151119. 
            doi"; pages: " 10.1073/pnas.2201151119. Epub 2022 Aug 5"
COMMENT     Sequence Label: pAAV-CamKII-DIO-hM4D(Gi)-mCherry
FEATURES             Location/Qualifiers
     source          1..7768
                     /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        145..1433
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     protein_bind    complement(1446..1479)
                     /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(1530..1563)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1572..2282)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1850..1869)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2119..2137
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2279..2288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3737..3746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3753..3786
                     /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    3837..3870
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3895..4483
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3948..3968)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4366..4377)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4515..4991
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4714..4733)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5031..5171
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5031..5160
                     /label=AAV2 ITR
     rep_origin      5246..5701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5333..5352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5543..5564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5718..5737)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5837..5859
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5897..5915)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5983..6087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6088..6156,6157..6948)
                     /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(6306..6325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7119..7707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7608..7627
                     /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 gcgtcattat ggccttaggt cacttcatct ccatggggtt
      181 cttcttctga ttttctagaa aatgagatgg gggtgcagag agcttcctca gtgacctgcc
      241 cagggtcaca tcagaaatgt cagagctaga acttgaactc agattactaa tcttaaattc
      301 catgccttgg gggcatgcaa gtacgatata cagaaggagt gaactcatta gggcagatga
      361 ccaatgagtt taggaaagaa gagtccaggg cagggtacat ctacaccacc cgcccagccc
      421 tgggtgagtc cagccacgtt cacctcatta tagttgcctc tctccagtcc taccttgacg
      481 ggaagcacaa gcagaaactg ggacaggagc cccaggagac caaatcttca tggtccctct
      541 gggaggatgg gtggggagag ctgtggcaga ggcctcagga ggggccctgc tgctcagtgg
      601 tgacagatag gggtgagaaa gcagacagag tcattccgtc agcattctgg gtctgtttgg
      661 tacttcttct cacgctaagg tggcggtgtg atatgcacaa tggctaaaaa gcagggagag
      721 ctggaaagaa acaaggacag agacagaggc caagtcaacc agaccaattc ccagaggaag
      781 caaagaaacc attacagaga ctacaagggg gaagggaagg agagatgaat tagcttcccc
      841 tgtaaacctt agaacccagc tgttgccagg gcaacggggc aatacctgtc tcttcagagg
      901 agatgaagtt gccagggtaa ctacatcctg tctttctcaa ggaccatccc agaatgtggc
      961 acccactagc cgttaccata gcaactgcct ctttgcccca cttaatccca tcccgtctgt
     1021 taaaagggcc ctatagttgg aggtggggga ggtaggaaga gcgatgatca cttgtggact
     1081 aagtttgttc gcatcccctt ctccaacccc ctcagtacat caccctgggg gaacagggtc
     1141 cacttgctcc tgggcccaca cagtcctgca gtattgtgta tataaggcca gggcaaagag
     1201 gagcaggttt taaagtgaaa ggcaggcagg tgttggggag gcagttaccg gggcaacggg
     1261 aacagggcgt ttcggaggtg gttgccatgg ggacctggat gctgacgaag gctcgcgagg
     1321 ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct cgtcagtcaa gccggttctc
     1381 cgtttgcact caggagcacg ggcaggcgag tggcccctag ttctgggggc agcgtcgact
     1441 ccggaataac ttcgtatagg atactttata cgaagttatg cagaatggta gctggattgt
     1501 agctgctatt agcaatatga aacctcttaa taacttcgta tagcatacat tatacgaagt
     1561 tatggcgcgc cttacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg
     1621 cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg
     1681 ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc
     1741 tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg
     1801 gcgccgtcct cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc
     1861 attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg
     1921 ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc
     1981 acgcgctccc acttgaagcc ctcggggaag gacagcttca agtagtcggg gatgtcggcg
     2041 gggtgcttca cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg
     2101 aagggcaggg ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg
     2161 cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc
     2221 accttgaagc gcatgaactc cttgatgatg gccatgttat cctcctcgcc cttgctcacc
     2281 atggtggcga ccggtggatc ccgcctggca gtgccgatgt tccgatactg gcacagcagc
     2341 aggtgccgga aggtcttttt aaaggtggcg ttgcacagag catagcaggc agggttgatg
     2401 gtgctgttga cgtagcagag ccagtagcca atggaccaca ccgtgtcagg gatgcagctc
     2461 tggcagaagg tgttcaccag gaccatgacg ttgtagggcg tccaggtgag gatgaaggcc
     2521 agcagaatgg caaagatcgt tcgtgtcact ttgcgctccc gggccgccat ctgccgcttc
     2581 ttgcgcacct ggttgcgagc gatgctggcg aacttgcggg ccacgttggc cgcagggcgc
     2641 atgccagccg gcgtggcagg cacaatctca atggctgtca cacactcatt gcctgtctgc
     2701 ttcgtcacaa tctggatctt ggaccatctg gaggctgggt tgagggcccg cggctgcagg
     2761 ggaggggcgg gcatggcggg cgtggtggcc tctgtggtgg acagctctgt ggctgggcgt
     2821 tccttggtgt tctgggtggc actgcctgag ctggactcat tggaagtgtc cttatcagcc
     2881 acggggcgcg gtggcggtgg cagcgctggc gggggggcct cctccagctt gccattgcgc
     2941 agctcctccc gggcggcctc cccgggcggg ggcttcttga cgctctgctt cattagtggg
     3001 ctcttgagga aggccagcgt cttggctttc ttctccttcg ggccctcggg ccggtgcttg
     3061 tggactcggc tgcgactggc cagggagatg tggatgtaca gcaccgtcat gatgaccaca
     3121 ggcaggtaga agccagcaat ggctgtgcca aaggtcactg ctgggttgga caggaactgg
     3181 atgaagcact ggttgtcggg caccgtccgc ttacccacca caaactgcca gaacaagatg
     3241 gcaggcgccc agagcacgaa ggacagtacc caggcagcag caatcatgag gcctgccatc
     3301 ttggtggtgc gccgggcagg gtaggtgaga ggcttggtga cgcagaagta gcggtcaaag
     3361 ctgatgatga gaaggttcat gacggaggcg ttgctcacca cgcagtccag ggccagccac
     3421 aggtcgcaga ccacggcgcc caggggccag tagcccttga tgatgtacac ggtgtagagg
     3481 ttcatggaga aggcgcctat gatgagatca gcacacgcca ggctgaagag gaagtagttg
     3541 ttgactgtct gcagctgcct gttgaccttg atggacagca tcaccaggat gttgcccacg
     3601 acagtcacca ggctcaggga gcctgtcact gtggcaatga agaccatttc caccgtctca
     3661 tagcgattgt gggatgatga cgtgaccagg cgcacggact gattgcccga gctgccattg
     3721 acaggtgtga agttggccat ggtggcgcta gcataacttc gtataaagta tcctatacga
     3781 agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa
     3841 cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat cgataatcaa
     3901 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3961 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4021 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4081 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4141 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4201 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4261 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     4321 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4381 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4441 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     4501 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     4561 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     4621 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     4681 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     4741 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     4801 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     4861 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     4921 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     4981 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     5041 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     5101 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     5161 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5221 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5281 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     5341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5461 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     5581 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     5641 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5701 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5761 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5821 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5881 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5941 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6001 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6061 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6121 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6181 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6241 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6301 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6361 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6421 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6481 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6541 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6601 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6661 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6721 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6781 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6841 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6901 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6961 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7021 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7081 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7141 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7201 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7261 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7321 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7381 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7441 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7501 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7561 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7621 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7681 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7741 gccttttgct ggccttttgc tcacatgt
//
