LOCUS       sequence-434486-        7252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing Chrimson-tdTomato under the control of E2 
            regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7252)
  AUTHORS   Vormstein-Schneider D, Lin JD, Pelkey KA, Chittajallu R, Guo B, 
            Arias-Garcia MA, Allaway K, Sakopoulos S, Schneider G, Stevenson O, 
            Vergara J, Sharma J, Zhang Q, Franken TP, Smith J, Ibrahim LA, M 
            Astro KJ, Sabri E, Huang S, Favuzzi E, Burbridge T, Xu Q, Guo L, 
            Vogel I, Sanchez V, Saldi GA, Gorissen BL, Yuan X, Zaghloul KA, 
            Devinsky O, Sabatini BL, Batista-Brito R, Reynolds J, Feng G, Fu Z, 
            McBain CJ, Fishell G, Dimidschstein J
  TITLE     Viral manipulation of functionally distinct interneurons in mice, 
            non-human primates and humans.
  JOURNAL   Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 
            10.1038/s41593-020-0692-9. Epub 2020 Aug 17.
  PUBMED    32807948
REFERENCE   2  (bases 1 to 7252)
  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/s41593-020-0692-9"; journalName: "Nat Neurosci"; date: 
            "2020-12"; volume: "23"; issue: "12"; pages: "1629-1636"
COMMENT     Sequence Label: pAAV_S5E2_Chrimson_tdTomato
FEATURES             Location/Qualifiers
     source          1..7252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          552..684
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   1488..1617
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1679..2267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1759..1778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2438..3229,3230..3298))
                     /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     3061..3080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3299..3403)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3471..3489
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3527..3549)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3649..3668
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3685..4140)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3822..3843)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4034..4053
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4215..4355
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4226..4355
                     /label=AAV2 ITR
     primer_bind     complement(4405..4424)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    4434..4515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4459..4478
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    4684..5272
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4790..4801
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5199..5219
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(6743..7252,1..540))
                     /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"
ORIGIN
        1 caggttgctc agtctgatca ggatcacggg gcaagacagc agccactcga agtagcgcag
       61 gcaataggcg tggttgccgg tagacaggta cactgtggcg gggctgctga actccttgaa
      121 gatctccagg gtcacgaaca gcacctcgac gcaacagacg tagacctcct cccaaccgca
      181 agtggccttc caggcgctga agccgtagaa tgtcagcagg gcgatggcga tgctgaaagc
      241 aatccactgg cagacctggg cgccgatctt ttctcctggt gtgcctctgc tgtgcagcca
      301 agcctgcttc ttctccactc ccacgaccag gtagccgcct gtggcctcca cgaagcaata
      361 tccgtatccg gcgtcgctca gtccgtaaga agggtcacac caccactcgg tgctgaagca
      421 ctcgcctgta ggtgtcattc ctccacagcc catgtcctcg tggtggtagg ggttaggcca
      481 agggttgatg cctccggcgg caaacagaga tctggtggcg ctgctgatca gctcagccat
      541 ggtggcggat cctgtggaga gaaaggcaaa gtggatgtca gtaagaccaa taggtgccta
      601 tcagaaacgc aagagtcttc tctgtctcga caagcccagt ttctattggt ctccttaaac
      661 ctgtcttgta accttgatac ttacgtgccc agtgcctcac gaccaacttc tgcagcttaa
      721 ggcgccgcgc tctgcttctg gaaggctgcg ctcctcgact tgcaggctag aagcaaatgt
      781 aagcaataga tggctctgcc ctgactttta tgcccagccc gggctgcagg aagtttaaaa
      841 taactctgag caattgagac taaatgcctc tgtatagaga aaagactgcc acagacagga
      901 atgttcttgg tgtctcttgg gtgcactgtt ccatcatacc atgtgtccca tatagtgctt
      961 tcatatagat tgtaaatact gcattatttt aaggaaatca tagttaatta aggtaagtat
     1021 ttccaaaatg gggttataac aatttactcc aacatcacat gttaatatga tctgagggaa
     1081 ttatgcatag tgtttgcctt ttgtttttgt ttatttttcc attactgttt catagcaatg
     1141 aagagacaaa tgctgactct agcagaattt cctcagtctg ctaagtatag ctacataatt
     1201 gccaccttca ccagcaccat tgggaaaaaa ataacctaca ttacttgact ccatatagct
     1261 atattgccat aactttgtag ggcttacagt aatgtggttt ctgtataaaa ctgaattgaa
     1321 atctcagaat tctacagaga gaggaatggg gacaagaaaa tgaggaggga actgaagtgg
     1381 cttgtgtcaa agccttcttt taggaagtgt gcacttaact tctagtcagt aagctatagc
     1441 agccatgtta gattaaacca ctagtttaat taaacgcgtg cggccgcagg aacccctagt
     1501 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     1561 ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca
     1621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1861 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2101 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2341 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2401 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2461 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2521 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2581 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2641 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2701 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2761 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2821 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2881 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2941 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3001 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3061 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3121 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3181 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3241 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3301 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3361 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3421 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3481 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3541 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3601 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     3661 agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt
     3721 taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt
     3781 ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc
     3841 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3901 gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3961 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4021 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag
     4081 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4141 actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4201 ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     4261 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     4321 agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgcat
     4381 gcggtatcga taagctgata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4441 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4501 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4561 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     4621 ctagagtcgc ggccagcttg atggccgcta tcacagatct ctcgagcagc gctcggtatc
     4681 gatgcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg
     4741 gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg
     4801 aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacatag gcgagcagcc
     4861 aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag
     4921 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg
     4981 ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg
     5041 ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct
     5101 cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     5161 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     5221 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa
     5281 gcttgatatc gaattcttac ttatacagct catccatgcc gtacagaaac agatggtggc
     5341 gtccttcaga cctctcatac tgttccacaa tggtgtagtc ttcattgtgg gatgtgatat
     5401 ccagcttggt gtccacatag taatagccgg gcagctgcac tggtttcttg gccatgtaga
     5461 tggttttgaa ctccaccagg tagtggccgc cgtctttcag cttcagggcc tggtggattt
     5521 cgcccttcag cactccgtct ctaggataca gccgttcagt gctggcctcc caccccattg
     5581 ttttcttctg catcacgggc ccatcgggtg gaaaattggt gccgcgcatt ttgactttgt
     5641 agatcagagt gccatcctgc agagaggagt cctgtgtcac ggtcaccagg cctccatctt
     5701 caaaattcat cactctctcc cacttaaagc cctctgggaa agacagtttc ttgtaatcag
     5761 gaatgtcagc gggatgcttg acataggcct tacttccgta cataaactga ggagacagga
     5821 tatcccaagc gaaaggcagt ggtcctccct tggtcacttt cagcttggca gtctgtgtgc
     5881 cctcgtaagg cctcccttcg ccctctcctt caatctcaaa ttcgtgccca ttcatgctgc
     5941 cctccatgcg caccttaaac ctcatgaact ctttgatcac agccatattg ttatcctcgc
     6001 ttgaggcggt tccactagac cctgagccgg tacttccagt cccatggccc aggaacagat
     6061 ggtgtctccc ctcgctccgt tcatactgct ccacaatagt gtagtcctcg ttgtgtgaag
     6121 tgatatccag ttttgtgtcc acgtaatagt agcctggcag ctgcacaggt ttcttggcca
     6181 tgtagattgt cttgaactcc accaggtaat gtcccccatc tttcagcttc agagcctggt
     6241 ggatttctcc tttcagcacc ccgtcgcgtg gatacagcct ttcggtggag gcctcccatc
     6301 ccatagtttt cttctgcatc acaggtccat cagggggaaa atttgtccct ctcattttca
     6361 ccttgtagat cagggtccca tcctgcaggg agctgtcctg ggtcacagtc accaggcctc
     6421 cgtcctcgaa gttcatcact ctttcccact taaacccctc ggggaagctc agtttcttat
     6481 aatcgggaat gtcagctgga tgtttcacat aggccttgga gccgtacata aactgtgggc
     6541 tcaggatgtc ccaagcgaat ggcagaggcc cgccctttgt cactttcagc ttggctgtct
     6601 gggtgccctc gtagggccgt ccctctcctt ctccctcaat ttcgaactcg tggccgttca
     6661 tgcttccttc cattctcacc ttaaaccgca tgaactcttt gatcacttcc tcgcccttac
     6721 tcactgctac taccggtgcc gccactgtgt cctcgtcctc ctcctccacg aactcttcca
     6781 cttccacctc ctcgcctccg atctccatct tggtggtctt ccggatgtcg ccgtggatca
     6841 ggatgtgctc gtggatcttg atcctcaggt ggtgggccag gaaggtccaa aactccttgg
     6901 cgatgatgtc gcagatgctg tggccgatgc tgttggcgta agggctcagc ttcagcagtc
     6961 cttctggtcc cactgcccag aggattgggt agctgcccca agaggcgaag taagcgtagg
     7021 ccatcagctt cacgaccatg cggcaatggc ctttaggcac gctgtggttg gcttccacgt
     7081 agcacttggc ggcctggaag tacatgtagc cgccgtagat gcaagacacg atatacagca
     7141 gccacttgag ccaatcggta gccagtcctg cggccatgcc gaacacgatc attcccacgc
     7201 aagacacgat caggcccatg gtccgcttgc tgtagtcgtt cttcaggccg ct
//
