LOCUS       sequence-257665-        7401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ChrimsonR-tdTomato under the CAG promoter
            in frt/reversed (Flp-dependent) manner. tdTomato has codons varied 
            to reduce recombination..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  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 7401)
  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/nmeth.2836"; journalName: "Nat Methods"; date: "2014-03"; 
            volume: "11"; issue: "3"; pages: "338-46"
COMMENT     Sequence Label: AAV-CAG-FLPX-rc [ChrimsonR-tdTomato]
FEATURES             Location/Qualifiers
     source          1..7401
                     /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        387..664
                     /label=chicken beta-actin promoter
     primer_bind     943..965
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1027..1046
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1079..1112
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(2652..3701)
                     /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"
     regulatory      3698..3707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3725..3758)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3848..4436
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3901..3921)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4319..4330)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   4664..4804
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4664..4793
                     /label=AAV2 ITR
     rep_origin      4879..5334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4966..4985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5176..5197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5351..5370)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5470..5492
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5530..5548)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5616..5720
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5721..5789,5790..6581)
                     /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(5939..5958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6752..7340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7241..7260
                     /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 gtcgacccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggggcg cgcgccrggg gggggggggg ggggsggggg
      541 ggggggsggg gsgrggcgga grggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
      721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
      841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggctgtc
      901 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
      961 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1021 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgctagcga
     1081 agttcctatt ctctagaaag tataggaact tcgcagaatg gtagctggat tgtagctgct
     1141 attagcaata tgaaacctct tagaagttcc tattcttcaa aaggtatagg aacttcgaat
     1201 tctcattact tatacagctc atccatgccg tacagaaaca gatggtggcg tccttcagac
     1261 ctctcatact gttccacaat ggtgtagtct tcattgtggg atgtgatatc cagcttggtg
     1321 tccacatagt aatagccggg cagctgcact ggtttcttgg ccatgtagat ggttttgaac
     1381 tccaccaggt agtggccgcc gtctttcagc ttcagggcct ggtggatttc gcccttcagc
     1441 actccgtctc taggatacag ccgttcagtg ctggcctccc accccattgt tttcttctgc
     1501 atcacgggcc catcgggtgg aaaattggtg ccgcgcattt tgactttgta gatcagagtg
     1561 ccatcctgca gagaggagtc ctgtgtcacg gtcaccaggc ctccatcttc aaaattcatc
     1621 actctctccc acttaaagcc ctctgggaaa gacagtttct tgtaatcagg aatgtcagcg
     1681 ggatgcttga cataggcctt acttccgtac ataaactgag gagacaggat atcccaagcg
     1741 aaaggcagtg gtcctccctt ggtcactttc agcttggcag tctgtgtgcc ctcgtaaggc
     1801 ctcccttcgc cctctccttc aatctcaaat tcgtgcccat tcatgctgcc ctccatgcgc
     1861 accttaaacc tcatgaactc tttgatcaca gccatattgt tatcctcgct tgaggcggtt
     1921 ccactagacc ctgagccggt acttccagtc ccatggccca ggaacagatg gtgtctcccc
     1981 tcgctccgtt catactgctc cacaatagtg tagtcctcgt tgtgtgaagt gatatccagt
     2041 tttgtgtcca cgtaatagta gcctggcagc tgcacaggtt tcttggccat gtagattgtc
     2101 ttgaactcca ccaggtaatg tcccccatct ttcagcttca gagcctggtg gatttctcct
     2161 ttcagcaccc cgtcgcgtgg atacagcctt tcggtggagg cctcccatcc catagttttc
     2221 ttctgcatca caggtccatc agggggaaaa tttgtccctc tcattttcac cttgtagatc
     2281 agggtcccat cctgcaggga gctgtcctgg gtcacagtca ccaggcctcc gtcctcgaag
     2341 ttcatcactc tttcccactt aaacccctcg gggaagctca gtttcttata atcgggaatg
     2401 tcagctggat gtttcacata ggccttggag ccgtacataa actgtgggct caggatgtcc
     2461 caagcgaatg gcagaggccc gccctttgtc actttcagct tggctgtctg ggtgccctcg
     2521 tagggccgtc cctctccttc tccctcaatt tcgaactcgt ggccgttcat gcttccttcc
     2581 attctcacct taaaccgcat gaactctttg atcacttcct cgcccttact cactgctact
     2641 accggtgccg ccactgtgtc ctcgtcctcc tcctccacga actcttccac ttccacctcc
     2701 tcgcctccga tctccatctt ggtggtcttc cggatgtcgc cgtggatcag gatgtgctcg
     2761 tggatcttga tcctcaggtg gtgggccagg aaggtccaaa actccttggc gatgatgtcg
     2821 cagatgctgt ggccgatgct gttggcgtaa gggctcagct tcagcagtcc ttctggtccc
     2881 actgcccaga ggattgggta gctgccccaa gaggcgaagt aagcgtaggc catcagcttc
     2941 acgaccatgc ggcaatggcc tttaggcacg ctgtggttgg cttccacgta gcacttggcg
     3001 gcctggaagt acatgtagcc gccgtagatg caagacacga tatacagcag ccacttgagc
     3061 caatcggtag ccagtcctgc ggccatgccg aacacgatca ttcccacgca agacacgatc
     3121 aggcccatgg tccgcttgct gtagtcgttc ttcaggccgc tcaggttgct cagtctgatc
     3181 aggatcacgg ggcaagacag cagccactcg aagtagcgca ggcaataggc gtggttgccg
     3241 gtagacaggt acactgtggc ggggctgctg aactccttga agatctccag ggtcacgaac
     3301 agcacctcga cgcaacagac gtagacctcc tcccaaccgc aagtggcctt ccaggcgctg
     3361 aagccgtaga atgtcagcag ggcgatggcg atgctgaaag caatccactg gcagacctgg
     3421 gcgccgatct tttctcctgg tgtgcctctg ctgtgcagcc aagcctgctt cttctccact
     3481 cccacgacca ggtagccgcc tgtggcctcc acgaagcaat atccgtatcc ggcgtcgctc
     3541 agtccgtaag aagggtcaca ccaccactcg gtgctgaagc actcgcctgt aggtgtcatt
     3601 cctccacagc ccatgtcctc gtggtggtag gggttaggcc aagggttgat gcctccggcg
     3661 gcaaacagag atctggtggc gctgctgatc agctcagcca tggtggcgga tccggcggcg
     3721 cgccgaagtt cctatacttt ctagagaata ggaacttctt gccttaaccc agaaattatc
     3781 actgttattc tttagaatgg tgcaaagaga agttcctata ccttttgaag aataggaact
     3841 tctccggaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3901 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3961 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4021 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4081 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4141 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4201 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4261 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4321 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4381 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgccttg
     4441 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4501 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4561 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4621 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcaggaacc cctagtgatg
     4681 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4741 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4801 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4861 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4921 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4981 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5041 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5101 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5161 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     5221 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     5281 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5341 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5401 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5461 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5521 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5581 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5641 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5701 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5761 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5821 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5881 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5941 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6001 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6061 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6121 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6181 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6241 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6301 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6361 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6421 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6481 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6541 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6601 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6661 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6721 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6781 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6841 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6901 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6961 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7021 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7081 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7141 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7201 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7261 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7321 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7381 gctggccttt tgctcacatg t
//
