LOCUS       sequence-169764-        6864 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV-mediated expression of ChrimsonR-tdTomato under the Syn 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6864)
  AUTHORS   Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK,
            Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, 
            Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, 
            Constantine-Paton M, Wong GK, Boyden ES
  TITLE     Independent optical excitation of distinct neural populations.
  JOURNAL   Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 
            2014 Feb 9.
  PUBMED    24509633
REFERENCE   2  (bases 1 to 6864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.2836"; journalName: "Nat Methods"; date: "2014-03"; 
            volume: "11"; issue: "3"; pages: "338-46"
COMMENT     Sequence Label: pAAV-Syn-ChrimsonR-tdT
FEATURES             Location/Qualifiers
     source          1..6864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     142..161
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   320..449
                     /label=AAV2 ITR
     promoter        549..996
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1009..1018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1015..2064
                     /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    3535..4123
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3588..3608)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4006..4017)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4164..4371
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4541..4670
                     /label=AAV2 ITR
     rep_origin      complement(4907..5495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4987..5006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5666..6457,6458..6526))
                     /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     6289..6308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6527..6631)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6657..6864,1..248))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6794..6815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
       61 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
      121 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
      181 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
      241 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      301 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      361 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      421 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      481 atctacgtag ccatgctcta ggaagatcgt aggttaatta atctagactg cagagggccc
      541 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
      601 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
      661 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
      721 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
      781 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
      841 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
      901 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
      961 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga gaggatccgc caccatggct
     1021 gagctgatca gcagcgccac cagatctctg tttgccgccg gaggcatcaa cccttggcct
     1081 aacccctacc accacgagga catgggctgt ggaggaatga cacctacagg cgagtgcttc
     1141 agcaccgagt ggtggtgtga cccttcttac ggactgagcg acgccggata cggatattgc
     1201 ttcgtggagg ccacaggcgg ctacctggtc gtgggagtgg agaagaagca ggcttggctg
     1261 cacagcagag gcacaccagg agaaaagatc ggcgcccagg tctgccagtg gattgctttc
     1321 agcatcgcca tcgccctgct gacattctac ggcttcagcg cctggaaggc cacttgcggt
     1381 tgggaggagg tctacgtctg ttgcgtcgag gtgctgttcg tgaccctgga gatcttcaag
     1441 gagttcagca gccccgccac agtgtacctg tctaccggca accacgccta ttgcctgcgc
     1501 tacttcgagt ggctgctgtc ttgccccgtg atcctgatca gactgagcaa cctgagcggc
     1561 ctgaagaacg actacagcaa gcggaccatg ggcctgatcg tgtcttgcgt gggaatgatc
     1621 gtgttcggca tggccgcagg actggctacc gattggctca agtggctgct gtatatcgtg
     1681 tcttgcatct acggcggcta catgtacttc caggccgcca agtgctacgt ggaagccaac
     1741 cacagcgtgc ctaaaggcca ttgccgcatg gtcgtgaagc tgatggccta cgcttacttc
     1801 gcctcttggg gcagctaccc aatcctctgg gcagtgggac cagaaggact gctgaagctg
     1861 agcccttacg ccaacagcat cggccacagc atctgcgaca tcatcgccaa ggagttttgg
     1921 accttcctgg cccaccacct gaggatcaag atccacgagc acatcctgat ccacggcgac
     1981 atccggaaga ccaccaagat ggagatcgga ggcgaggagg tggaagtgga agagttcgtg
     2041 gaggaggagg acgaggacac agtggcggca ccggtagtag cagtgagtaa gggcgaggaa
     2101 gtgatcaaag agttcatgcg gtttaaggtg agaatggaag gaagcatgaa cggccacgag
     2161 ttcgaaattg agggagaagg agagggacgg ccctacgagg gcacccagac agccaagctg
     2221 aaagtgacaa agggcgggcc tctgccattc gcttgggaca tcctgagccc acagtttatg
     2281 tacggctcca aggcctatgt gaaacatcca gctgacattc ccgattataa gaaactgagc
     2341 ttccccgagg ggtttaagtg ggaaagagtg atgaacttcg aggacggagg cctggtgact
     2401 gtgacccagg acagctccct gcaggatggg accctgatct acaaggtgaa aatgagaggg
     2461 acaaattttc cccctgatgg acctgtgatg cagaagaaaa ctatgggatg ggaggcctcc
     2521 accgaaaggc tgtatccacg cgacggggtg ctgaaaggag aaatccacca ggctctgaag
     2581 ctgaaagatg ggggacatta cctggtggag ttcaagacaa tctacatggc caagaaacct
     2641 gtgcagctgc caggctacta ttacgtggac acaaaactgg atatcacttc acacaacgag
     2701 gactacacta ttgtggagca gtatgaacgg agcgagggga gacaccatct gttcctgggc
     2761 catgggactg gaagtaccgg ctcagggtct agtggaaccg cctcaagcga ggataacaat
     2821 atggctgtga tcaaagagtt catgaggttt aaggtgcgca tggagggcag catgaatggg
     2881 cacgaatttg agattgaagg agagggcgaa gggaggcctt acgagggcac acagactgcc
     2941 aagctgaaag tgaccaaggg aggaccactg cctttcgctt gggatatcct gtctcctcag
     3001 tttatgtacg gaagtaaggc ctatgtcaag catcccgctg acattcctga ttacaagaaa
     3061 ctgtctttcc cagagggctt taagtgggag agagtgatga attttgaaga tggaggcctg
     3121 gtgaccgtga cacaggactc ctctctgcag gatggcactc tgatctacaa agtcaaaatg
     3181 cgcggcacca attttccacc cgatgggccc gtgatgcaga agaaaacaat ggggtgggag
     3241 gccagcactg aacggctgta tcctagagac ggagtgctga agggcgaaat ccaccaggcc
     3301 ctgaagctga aagacggcgg ccactacctg gtggagttca aaaccatcta catggccaag
     3361 aaaccagtgc agctgcccgg ctattactat gtggacacca agctggatat cacatcccac
     3421 aatgaagact acaccattgt ggaacagtat gagaggtctg aaggacgcca ccatctgttt
     3481 ctgtacggca tggatgagct gtataagtaa gaattcgata tcaagcttat cgataatcaa
     3541 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3601 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3661 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3721 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3781 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3841 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3901 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3961 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4021 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4081 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc
     4141 gggcggccgc tcataaagct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4201 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4261 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4321 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtccg
     4381 gactgtacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     4441 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc
     4501 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     4561 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4621 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat
     4681 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4741 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4801 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     6661 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     6721 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     6781 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     6841 caaagggcga aaaaccgtct atca
//
