LOCUS       sequence-238046-        7341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""Activate neurons with 2p stimulation"".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  AUTHORS   Daie K, Svoboda K, Druckmann S
  TITLE     Targeted photostimulation uncovers circuit motifs supporting 
            short-term memory.
  JOURNAL   Nat Neurosci. 2021 Feb;24(2):259-265. doi: 
            10.1038/s41593-020-00776-3. Epub 2021 Jan 25.
  PUBMED    33495637
REFERENCE   2  (bases 1 to 7341)
  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"; doi: 
            "10.1038/s41593-020-00776-3"; journalName: "Nat Neurosci"; date: 
            "2021-02"; volume: "24"; issue: "2"; pages: "259-265"
COMMENT     Sequence Label: pAAV-CamKIIa-ChrimsonR::FusionRed::Kv2.1
FEATURES             Location/Qualifiers
     source          1..7341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     161..180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   322..451
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        470..1758
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1791..1800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1797..2846
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=ChrimsonR
                     /note="K176R mutant with accelerated closing kinetics"
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     misc_feature    3789..4377
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3842..3862)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4260..4271)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4409..4885
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4608..4627)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4925..5065
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4925..5054
                     /label=AAV2 ITR
     rep_origin      5140..5595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5227..5246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5437..5458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5612..5631)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5731..5753
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5791..5809)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5877..5981
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5982..6050,6051..6842)
                     /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(6200..6219)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7013..7341,1..260)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
       61 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      301 gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg
      361 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      421 ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct
      481 taggtcactt catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg
      541 cagagagctt cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg
      601 aactcagatt actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa
      661 ggagtgaact cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg
      721 tacatctaca ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt
      781 gcctctctcc agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag
      841 gagaccaaat cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct
      901 caggaggggc cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt
      961 ccgtcagcat tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg
     1021 cacaatggct aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt
     1081 caaccagacc aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg
     1141 gaaggagaga tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac
     1201 ggggcaatac ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt
     1261 ctcaaggacc atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg
     1321 ccccacttaa tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag
     1381 gaagagcgat gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag
     1441 tacatcaccc tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt
     1501 gtgtatataa ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg
     1561 gggaggcagt taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc
     1621 tggatgctga cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc
     1681 aagctcgtca gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc
     1741 cctagttctg ggggcagctc tagagcggta ccggatcccc gggtaccggt gccaccatgg
     1801 ctgagctcat ttcatctgct acacgctctc tgtttgccgc cgggggaata aacccttggc
     1861 ctaatcctta ccaccacgaa gacatgggtt gtggaggaat gacccccact ggagaatgtt
     1921 tctctaccga gtggtggtgt gatccttcat atggtctcag cgatgctggt tacggctatt
     1981 gtttcgtgga agctactgga ggctatctgg ttgttggggt cgagaaaaaa caggcttggc
     2041 ttcactcccg gggcacccca ggagagaaaa ttggagccca ggtatgtcag tggattgcct
     2101 tttctatagc aatcgccctc cttacatttt atgggttttc agcatggaag gctacatgcg
     2161 gttgggaaga ggtttacgtc tgttgtgtcg aagttttgtt cgtcaccctc gagatcttta
     2221 aagagttttc ttcaccagct acagtctacc tttctactgg caaccatgca tactgcctgc
     2281 gatatttcga gtggctcctg tcttgccccg tcatactcat tcgcctttct aacttgagtg
     2341 ggcttaagaa cgactacagt aaaagaacta tgggtcttat cgtgtcatgt gtcggcatga
     2401 tagtgttcgg gatggccgcc ggcctcgcta ccgattggtt gaaatggctc ctttatatcg
     2461 tttcttgcat ttacggaggg tacatgtatt tccaggcagc caaatgttat gttgaagcca
     2521 atcattccgt accaaaaggc cactgtcgaa tggtagttaa gctgatggct tacgcttact
     2581 ttgcaagttg gggatcatat ccaatactgt gggcagtggg gcctgaaggc ttgcttaagc
     2641 tctctccata tgctaactct ataggtcatt caatctgtga cataatagca aaagaatttt
     2701 ggaccttcct cgctcatcat ctgcgcatca agatccatga gcatatcctg atacacggtg
     2761 acatacgaaa gaccaccaaa atggaaattg gcggagaaga ggtggaggta gaggagtttg
     2821 ttgaagagga agacgaagat actgttgcag caccagttgt tgctgtttct gagctcatca
     2881 aagaaaacat gccaatgaag ttgtacatgg agggtactgt caacaatcac cattttaagt
     2941 gtaccagcga gggggagggg aagccctatg aagggactca gaccatgagg ataaaagttg
     3001 tagaaggggg gccccttcct ttcgcctttg acattctcgc cacatctttt atgtatggta
     3061 gccgcacctt cattaagcat ccccctggga ttcccgattt ctttaaacaa tcttttccag
     3121 aaggcttcac ttgggaaagg gtaactactt atgaggacgg cggagtgctt acagccaccc
     3181 aagatacctc tctgcaggat ggctgcttga tatacaacgt aaaggtacgg ggcgttaact
     3241 tccctgcaaa tggtcctgtt atgcaaaaga aaaccctcgg atgggaggcc tctactgaga
     3301 ccatgtaccc tgcagatggg ggccttgagg gggcatgtga tatggcactt aaattggtag
     3361 gtggtggaca cttgatatgc aacctggaga ctacttaccg gagcaaaaaa cccgcaacca
     3421 atctcaaaat gccaggcgtt tacaacgtgg accatcgact tgagcggata aaggaggccg
     3481 acgatgagac ttatgttgag cagcatgagg tcgctgtagc taggtactca accggcgggg
     3541 ccggggatgg tggtaagtac aaacaatctc agcctatctt gaatactaag gagatggctc
     3601 cacagagtaa accaccagaa gagttggaga tgtcatccat gccatcacca gtggcacccc
     3661 tcccagccag aaccgagggc gtgattgaca tgcgatccat gagttcaata gatagcttca
     3721 ttagttgtgc aactgacttt cccgaagcaa ccagattcaa gtaagaattc gatatcaagc
     3781 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3841 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3901 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3961 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     4021 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccggaact ttcgctttcc
     4081 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4141 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     4201 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     4261 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     4321 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     4381 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     4441 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     4501 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     4561 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     4621 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4681 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4741 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4801 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4861 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4921 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4981 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     5041 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     5101 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     5161 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     5221 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5281 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5341 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5401 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5461 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     5521 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5581 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5641 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5701 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5761 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5821 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5881 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5941 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6001 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6061 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6121 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6181 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6241 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6301 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6361 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6421 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6481 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6541 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6601 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6661 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6721 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6781 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6841 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6901 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6961 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7021 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7081 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7141 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7201 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7261 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7321 ggtcgggctg aacggggggt t
//
