LOCUS       sequence-257684-        6954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ChrimsonR-tdTomato under the Syn 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 6954)
  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 6954)
  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-Syn-FLPX-rc [ChrimsonR-tdTomato]
FEATURES             Location/Qualifiers
     source          1..6954
                     /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        172..619
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    632..665
                     /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(2205..3254)
                     /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      3251..3260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3278..3311)
                     /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    3401..3989
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3454..3474)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3872..3883)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   4217..4357
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4217..4346
                     /label=AAV2 ITR
     rep_origin      4432..4887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4519..4538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4729..4750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4904..4923)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5023..5045
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5083..5101)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5169..5273
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5274..5342,5343..6134)
                     /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(5492..5511)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6305..6893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6794..6813
                     /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 gtcgacttaa ttaatctaga ctgcagaggg ccctgcgtat gagtgcaagt
      181 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc
      241 actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag
      301 gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc
      361 ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc
      421 actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc
      481 cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct
      541 gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt
      601 gtcgtgcctg agagcgcagt cgagagctag cgaagttcct attctctaga aagtatagga
      661 acttcgcaga atggtagctg gattgtagct gctattagca atatgaaacc tcttagaagt
      721 tcctattctt caaaaggtat aggaacttcg aattctcatt acttatacag ctcatccatg
      781 ccgtacagaa acagatggtg gcgtccttca gacctctcat actgttccac aatggtgtag
      841 tcttcattgt gggatgtgat atccagcttg gtgtccacat agtaatagcc gggcagctgc
      901 actggtttct tggccatgta gatggttttg aactccacca ggtagtggcc gccgtctttc
      961 agcttcaggg cctggtggat ttcgcccttc agcactccgt ctctaggata cagccgttca
     1021 gtgctggcct cccaccccat tgttttcttc tgcatcacgg gcccatcggg tggaaaattg
     1081 gtgccgcgca ttttgacttt gtagatcaga gtgccatcct gcagagagga gtcctgtgtc
     1141 acggtcacca ggcctccatc ttcaaaattc atcactctct cccacttaaa gccctctggg
     1201 aaagacagtt tcttgtaatc aggaatgtca gcgggatgct tgacataggc cttacttccg
     1261 tacataaact gaggagacag gatatcccaa gcgaaaggca gtggtcctcc cttggtcact
     1321 ttcagcttgg cagtctgtgt gccctcgtaa ggcctccctt cgccctctcc ttcaatctca
     1381 aattcgtgcc cattcatgct gccctccatg cgcaccttaa acctcatgaa ctctttgatc
     1441 acagccatat tgttatcctc gcttgaggcg gttccactag accctgagcc ggtacttcca
     1501 gtcccatggc ccaggaacag atggtgtctc ccctcgctcc gttcatactg ctccacaata
     1561 gtgtagtcct cgttgtgtga agtgatatcc agttttgtgt ccacgtaata gtagcctggc
     1621 agctgcacag gtttcttggc catgtagatt gtcttgaact ccaccaggta atgtccccca
     1681 tctttcagct tcagagcctg gtggatttct cctttcagca ccccgtcgcg tggatacagc
     1741 ctttcggtgg aggcctccca tcccatagtt ttcttctgca tcacaggtcc atcaggggga
     1801 aaatttgtcc ctctcatttt caccttgtag atcagggtcc catcctgcag ggagctgtcc
     1861 tgggtcacag tcaccaggcc tccgtcctcg aagttcatca ctctttccca cttaaacccc
     1921 tcggggaagc tcagtttctt ataatcggga atgtcagctg gatgtttcac ataggccttg
     1981 gagccgtaca taaactgtgg gctcaggatg tcccaagcga atggcagagg cccgcccttt
     2041 gtcactttca gcttggctgt ctgggtgccc tcgtagggcc gtccctctcc ttctccctca
     2101 atttcgaact cgtggccgtt catgcttcct tccattctca ccttaaaccg catgaactct
     2161 ttgatcactt cctcgccctt actcactgct actaccggtg ccgccactgt gtcctcgtcc
     2221 tcctcctcca cgaactcttc cacttccacc tcctcgcctc cgatctccat cttggtggtc
     2281 ttccggatgt cgccgtggat caggatgtgc tcgtggatct tgatcctcag gtggtgggcc
     2341 aggaaggtcc aaaactcctt ggcgatgatg tcgcagatgc tgtggccgat gctgttggcg
     2401 taagggctca gcttcagcag tccttctggt cccactgccc agaggattgg gtagctgccc
     2461 caagaggcga agtaagcgta ggccatcagc ttcacgacca tgcggcaatg gcctttaggc
     2521 acgctgtggt tggcttccac gtagcacttg gcggcctgga agtacatgta gccgccgtag
     2581 atgcaagaca cgatatacag cagccacttg agccaatcgg tagccagtcc tgcggccatg
     2641 ccgaacacga tcattcccac gcaagacacg atcaggccca tggtccgctt gctgtagtcg
     2701 ttcttcaggc cgctcaggtt gctcagtctg atcaggatca cggggcaaga cagcagccac
     2761 tcgaagtagc gcaggcaata ggcgtggttg ccggtagaca ggtacactgt ggcggggctg
     2821 ctgaactcct tgaagatctc cagggtcacg aacagcacct cgacgcaaca gacgtagacc
     2881 tcctcccaac cgcaagtggc cttccaggcg ctgaagccgt agaatgtcag cagggcgatg
     2941 gcgatgctga aagcaatcca ctggcagacc tgggcgccga tcttttctcc tggtgtgcct
     3001 ctgctgtgca gccaagcctg cttcttctcc actcccacga ccaggtagcc gcctgtggcc
     3061 tccacgaagc aatatccgta tccggcgtcg ctcagtccgt aagaagggtc acaccaccac
     3121 tcggtgctga agcactcgcc tgtaggtgtc attcctccac agcccatgtc ctcgtggtgg
     3181 taggggttag gccaagggtt gatgcctccg gcggcaaaca gagatctggt ggcgctgctg
     3241 atcagctcag ccatggtggc ggatccggcg gcgcgccgaa gttcctatac tttctagaga
     3301 ataggaactt cttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     3361 agaagttcct ataccttttg aagaatagga acttctccgg aatcaacctc tggattacaa
     3421 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3481 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3541 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3601 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3661 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3721 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3781 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3841 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3901 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3961 tcggatctcc ctttgggccg cctccccgcc ttgtgccttc tagttgccag ccatctgttg
     4021 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4081 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4141 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     4201 cggtgggctc tatggcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4261 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4321 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4381 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4441 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4501 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4561 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4621 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4681 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4741 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     4801 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4861 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4921 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4981 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5041 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5101 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5161 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5221 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5281 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5341 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5401 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5461 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5521 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5581 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5641 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5701 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5761 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5821 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5881 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5941 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6001 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6061 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6121 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6181 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6241 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6301 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6361 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6421 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6481 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6541 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6601 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6661 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6721 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6781 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6841 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6901 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
