LOCUS       sequence-371327-        6310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry flanked by DIO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  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 6310)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..6310
                     /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"
     protein_bind    2295..2328
                     /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    2379..2412
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2437..3025
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2490..2510)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2908..2919)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3057..3533
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3256..3275)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3573..3713
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3573..3702
                     /label=AAV2 ITR
     rep_origin      3788..4243
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3875..3894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4085..4106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4260..4279)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4379..4401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4439..4457)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4525..4629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4630..4698,4699..5490)
                     /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(4848..4867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5661..6249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6150..6169
                     /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 atggtggcgc tagcataact tcgtataaag tatcctatac gaagttattt gccttaaccc
     2341 agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct
     2401 atacgaagtt atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     2461 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2521 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2581 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2641 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2701 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2761 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2821 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     2881 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2941 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3001 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     3061 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     3121 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     3181 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     3241 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     3301 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3361 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3421 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3481 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3541 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3601 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3661 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3721 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3781 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3841 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3901 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3961 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4021 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4081 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4141 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4201 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4261 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4321 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4381 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4441 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4501 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4561 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4621 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4681 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4741 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4801 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4861 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4921 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4981 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5041 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5101 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5161 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5221 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5281 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5341 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5401 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5461 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5521 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5581 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6301 gctcacatgt
//
