LOCUS       sequence-242020-        7325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Soma-localized ChrimsonR for dense expression in excitatory neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7325)
  AUTHORS   Chettih SN, Harvey CD
  TITLE     Single-neuron perturbations reveal feature-specific competition in 
            V1.
  JOURNAL   Nature. 2019 Mar 6. pii: 10.1038/s41586-019-0997-6. doi: 
            10.1038/s41586-019-0997-6.
  PUBMED    30842660
REFERENCE   2  (bases 1 to 7325)
  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: "Nature. 
            2019 Mar 6. pii: 10.1038/s41586-019-0997-6. doi: 
            10.1038/s41586-019-0997-6."
COMMENT     Sequence Label: pAAV-CamKIIa-ChrimsonR-mScarlet-KV2.1
FEATURES             Location/Qualifiers
     source          1..7325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   80..209
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        228..1516
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1551..1560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1557..2606
                     /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"
     CDS             2811..3503
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2941..2959)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3209..3228
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3531..4119
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3584..3604)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4002..4013)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4151..4627
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4350..4369)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4667..4807
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4667..4796
                     /label=AAV2 ITR
     rep_origin      4882..5337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4969..4988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5179..5200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5354..5373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5473..5495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5533..5551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5619..5723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5724..5792,5793..6584)
                     /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(5942..5961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6755..7325,1..18)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7244..7263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
       61 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
      121 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      181 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtttaacat tatggcctta
      241 ggtcacttca tctccatggg gttcttcttc tgattttcta gaaaatgaga tgggggtgca
      301 gagagcttcc tcagtgacct gcccagggtc acatcagaaa tgtcagagct agaacttgaa
      361 ctcagattac taatcttaaa ttccatgcct tgggggcatg caagtacgat atacagaagg
      421 agtgaactca ttagggcaga tgaccaatga gtttaggaaa gaagagtcca gggcagggta
      481 catctacacc acccgcccag ccctgggtga gtccagccac gttcacctca ttatagttgc
      541 ctctctccag tcctaccttg acgggaagca caagcagaaa ctgggacagg agccccagga
      601 gaccaaatct tcatggtccc tctgggagga tgggtgggga gagctgtggc agaggcctca
      661 ggaggggccc tgctgctcag tggtgacaga taggggtgag aaagcagaca gagtcattcc
      721 gtcagcattc tgggtctgtt tggtacttct tctcacgcta aggtggcggt gtgatatgca
      781 caatggctaa aaagcaggga gagctggaaa gaaacaagga cagagacaga ggccaagtca
      841 accagaccaa ttcccagagg aagcaaagaa accattacag agactacaag ggggaaggga
      901 aggagagatg aattagcttc ccctgtaaac cttagaaccc agctgttgcc agggcaacgg
      961 ggcaatacct gtctcttcag aggagatgaa gttgccaggg taactacatc ctgtctttct
     1021 caaggaccat cccagaatgt ggcacccact agccgttacc atagcaactg cctctttgcc
     1081 ccacttaatc ccatcccgtc tgttaaaagg gccctatagt tggaggtggg ggaggtagga
     1141 agagcgatga tcacttgtgg actaagtttg ttcgcatccc cttctccaac cccctcagta
     1201 catcaccctg ggggaacagg gtccacttgc tcctgggccc acacagtcct gcagtattgt
     1261 gtatataagg ccagggcaaa gaggagcagg ttttaaagtg aaaggcaggc aggtgttggg
     1321 gaggcagtta ccggggcaac gggaacaggg cgtttcggag gtggttgcca tggggacctg
     1381 gatgctgacg aaggctcgcg aggctgtgag cagccacagt gccctgctca gaagccccaa
     1441 gctcgtcagt caagccggtt ctccgtttgc actcaggagc acgggcaggc gagtggcccc
     1501 tagttctggg ggcagctcta gagcggtacc ggatccccgg gtaccggtag gccaccatgg
     1561 ctgagctgat cagcagcgcc accagatctc tgtttgccgc cggaggcatc aacccttggc
     1621 ctaaccccta ccaccacgag gacatgggct gtggaggaat gacacctaca ggcgagtgct
     1681 tcagcaccga gtggtggtgt gacccttctt acggactgag cgacgccgga tacggatatt
     1741 gcttcgtgga ggccacaggc ggctacctgg tcgtgggagt ggagaagaag caggcttggc
     1801 tgcacagcag aggcacacca ggagaaaaga tcggcgccca ggtctgccag tggattgctt
     1861 tcagcatcgc catcgccctg ctgacattct acggcttcag cgcctggaag gccacttgcg
     1921 gttgggagga ggtctacgtc tgttgcgtcg aggtgctgtt cgtgaccctg gagatcttca
     1981 aggagttcag cagccccgcc acagtgtacc tgtctaccgg caaccacgcc tattgcctgc
     2041 gctacttcga gtggctgctg tcttgccccg tgatcctgat cagactgagc aacctgagcg
     2101 gcctgaagaa cgactacagc aagcggacca tgggcctgat cgtgtcttgc gtgggaatga
     2161 tcgtgttcgg catggccgca ggactggcta ccgattggct caagtggctg ctgtatatcg
     2221 tgtcttgcat ctacggcggc tacatgtact tccaggccgc caagtgctac gtggaagcca
     2281 accacagcgt gcctaaaggc cattgccgca tggtcgtgaa gctgatggcc tacgcttact
     2341 tcgcctcttg gggcagctac ccaatcctct gggcagtggg accagaagga ctgctgaagc
     2401 tgagccctta cgccaacagc atcggccaca gcatctgcga catcatcgcc aaggagtttt
     2461 ggaccttcct ggcccaccac ctgaggatca agatccacga gcacatcctg atccacggcg
     2521 acatccggaa gaccaccaag atggagatcg gaggcgagga ggtggaagtg gaagagttcg
     2581 tggaggagga ggacgaggac acagtggcgg ccgcccagtc ccagcccatc ctcaacacca
     2641 aggagatggc cccgcagagc aagcctccag aggagctgga gatgagcagc atgcccagcc
     2701 ccgtggcccc tctgcccgca cgcacggagg gcgtcatcga catgcggagc atgtccagca
     2761 ttgacagctt catcagctgt gccacggact tccctgaagc caccagattc gtgagcaagg
     2821 gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc tccatgaacg
     2881 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2941 ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc ctgggacatc ctgtcccctc
     3001 agttcatgta cggctccagg gccttcacca agcaccccgc cgacatcccc gactactata
     3061 agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     3121 ccgtgaccgt gacccaggac acctccctgg aggacggcac cctgatctac aaggtgaagc
     3181 tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca atgggctggg
     3241 aagcgtccac cgagcggttg taccccgagg acggcgtgct gaagggcgac attaagatgg
     3301 ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc tacaaggcca
     3361 agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg caagttggac atcacctccc
     3421 acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgagggccgc cactccaccg
     3481 gcggcatgga cgagctgtac aagtaagaat tcgatatcaa gcttatcgat aatcaacctc
     3541 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3601 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3661 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3721 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3781 ttgccaccac ctgtcagctc ctttccggaa ctttcgcttt ccccctccct attgccacgg
     3841 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3901 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     3961 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4021 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4081 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     4141 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     4201 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     4261 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     4321 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     4381 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     4441 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     4501 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     4561 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     4621 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     4681 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4741 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4801 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4861 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4921 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     4981 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5041 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     5101 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5161 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     5221 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     5281 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     5341 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5401 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5461 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5521 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5581 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5641 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5701 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5761 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5821 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5881 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5941 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6001 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6061 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6121 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6181 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6241 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6301 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6361 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6421 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6481 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6541 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6601 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6661 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6721 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6841 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7201 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7261 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7321 caggg
//
