LOCUS       sequence-220097-        7144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a Cre-dependent red light-shifted channelrhodopsin 
            variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7144)
  AUTHORS   Messier JE, Chen H, Cai ZL, Xue M
  TITLE     Targeting light-gated chloride channels to neuronal somatodendritic 
            domain reduces their excitatory effect in the axon.
  JOURNAL   Elife. 2018 Aug 9;7. pii: 38506. doi: 10.7554/eLife.38506.
  PUBMED    30091701
REFERENCE   2  (bases 1 to 7144)
  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: "Elife."; 
            date: "2018-08-9"; volume: "7. pii: 38506. doi"; pages: " 
            10.7554/eLife.38506"
COMMENT     Sequence Label: pAAV-EF1-alpha-DIO-ReaChR-P2A-dTomato
FEATURES             Location/Qualifiers
     source          1..7144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(35..68)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(119..152)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      162..171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1233..1289
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1290..1991
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1423..1441)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1691..1710
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1885..1908
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    2009..2042
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2093..2126
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(2157..3335)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2166..3104
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(2189..2209)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   3436..3565
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3627..4215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3707..3726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4386..5177,5178..5246))
                     /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     5009..5028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5247..5351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5419..5437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5475..5497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5597..5616
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5633..6088)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5770..5791)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5982..6001
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6163..6303
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6174..6303
                     /label=AAV2 ITR
     polyA_signal    6421..6532
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    join(6566..7144,1..10)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6672..6683
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7081..7101
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 gaggttgatt atcgataagc ttgatatcga attcataact tcgtatagca tacattatac
       61 gaagttattc tttgcaccat tctaaagaat aacagtgata atttctgggt taaggcaaat
      121 aacttcgtat aggatacttt atacgaagtt atgctagcgg cgccaccatg gtgagcagaa
      181 gaccctggct gctggccctg gccctggccg tggccctggc cgccggcagc gccggcgcca
      241 gcaccggcag cgacgccacc gtgcccgtgg ccacccagga cggccccgac tacgtgttcc
      301 acagagccca cgagagaatg ctgttccaga ccagctacac cctggagaac aacggcagcg
      361 tgatctgcat ccccaacaac ggccagtgct tctgcctggc ctggctgaag agtaacggca
      421 ccaacgccga gaagctggcc gccaacatcc tgcagtgggt ggtgtttgcg ctgagcgtgg
      481 cgtgcctggg ctggtatgcg tatcaggcgt ggcgcgcgac ctgcggctgg gaagaagtgt
      541 atgtggcgct gattgaaatg atgaaaagca ttattgaagc gtttcatgaa tttgatagcc
      601 cggcgaccct gtggctgagc agcggcaacg gcgtggtgtg gatgcgctat ggcgaatggc
      661 tgctgacctg ccccgtgatt ctgattcatc tgagcaacct gaccggcctg aaagatgatt
      721 atagcaaacg caccatgggc ctgctggtga gcgacgtggg ctgcattgtg tggggcgcga
      781 ccagcgcgat gtgcaccggc tggaccaaaa ttctgttttt tctgattagc ctgagctatg
      841 gcatgtatac ctattttcat gcggccaaag tgtatattga agcgtttcat accgtgccga
      901 aaggcctgtg cagacagctg gtgagagcca tggcctggct gttcttcgtg agctggggca
      961 tgttccccgt gctgttcctg ctgggccccg agggcttcgg ccatattagc ccgtatggca
     1021 gcgcgattgg ccatagcatt ctggatctga ttgcgaagaa catgtggggc gtgctgggca
     1081 actatctgcg cgtgaaaatt catgaacata ttctgctgta tggcgatatt cgcaaaaaac
     1141 agaaaattac cattgcgggc caggaaatgg aagtggaaac cctggtggcg gaagaagaag
     1201 ataagtacga gagcagcgtt aacggaagcg gagccactaa cttctccctg ttgaaacaag
     1261 caggggatgt cgaagagaat cccgggccaa tggtgagcaa gggcgaggag gtcatcaaag
     1321 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     1381 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1441 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca
     1501 aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg
     1561 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg
     1621 actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc
     1681 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     1741 tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg
     1801 gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc
     1861 ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     1921 tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac ggcatggacg
     1981 agctgtacaa gtaagtcgac ggcgcgccat aacttcgtat aatgtatgct atacgaagtt
     2041 attaagaggt ttcatattgc taatagcagc tacaatccag ctaccattct gcataacttc
     2101 gtataaagta tcctatacga agttattccg gagtcgactc tagaggatcc ggtacctcac
     2161 gacacctgaa atggaagaaa aaaactttga accactgtct gaggcttgag aatgaaccaa
     2221 gatccaaact caaaaagggc aaattccaag gagaattaca tcaagtgcca agctggccta
     2281 acttcagtct ccacccactc agtgtgggga aactccatcg cataaaaccc ctccccccaa
     2341 cctaaagacg acgtactcca aaagctcgag aactaatcga ggtgcctgga cggcgcccgg
     2401 tactccgtgg agtcacatga agcgacggct gaggacggaa aggccctttt cctttgtgtg
     2461 ggtgactcac ccgcccgctc tcccgagcgc cgcgtcctcc attttgagct ccctgcagca
     2521 gggccgggaa gcggccatct ttccgctcac gcaactggtg ccgaccgggc cagccttgcc
     2581 gcccagggcg gggcgataca cggcggcgcg aggccaggca ccagagcagg ccggccagct
     2641 tgagactacc cccgtccgat tctcggtggc cgcgctcgca ggccccgcct cgccgaacat
     2701 gtgcgctggg acgcacgggc cccgtcgccg cccgcggccc caaaaaccga aataccagtg
     2761 tgcagatctt ggcccgcatt tacaagacta tcttgccaga aaaaaagcgt cgcagcaggt
     2821 catcaaaaat tttaaatggc tagagactta tcgaaagcag cgagacaggc gcgaaggtgc
     2881 caccagattc gcacgcggcg gccccagcgc ccaggccagg cctcaactca agcacgaggc
     2941 gaaggggctc cttaagcgca aggcctcgaa ctctcccacc cacttccaac ccgaagctcg
     3001 ggatcaagaa tcacgtactg cagccaggtg gaagtaattc aaggcacgca agggccataa
     3061 cccgtaaaga ggccaggccc gcgggaacca cacacggcac ttacctgtgt tctggcggca
     3121 aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg ttctccccca
     3181 ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac cccgcgccac
     3241 cttctctagg caccggttca attgccgacc cctcccccca acttctcggg gactgtgggc
     3301 gatgtgcgct ctgcccactg acgggcaccg gagccaattc ccactccttt caagacctag
     3361 aaggtccatt agctgcaaag attcctctct gtttaaaact ttatccatct ttgcaaagct
     3421 tacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3481 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc
     3541 gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac
     3601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5401 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5461 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5521 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5581 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta
     5641 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5701 caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg
     5761 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5821 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt
     5881 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5941 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6001 gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6061 gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa
     6121 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg
     6181 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     6241 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     6301 cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc ccaatcctcc
     6361 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     6421 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     6481 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaagat
     6541 ctctcgagca gcgctcggta tcgatgcggg gaggcggccc aaagggagat ccgactcgtc
     6601 tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc gcgggaagga
     6661 aggtccgctg gattgagggc cgaagggacg tagcagaagg acgtcccgcg cagaatccag
     6721 gtggcaacat aggcgagcag ccaaggaaag gacgatgatt tccccgacaa caccacggaa
     6781 ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt
     6841 tccgccgtgg caatagggag ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg
     6901 gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag tgcacaccac gccacgttgc
     6961 ctgacaacgg gccacaactc ctcataaaga gacagcaacc aggatttata caaggaggag
     7021 aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca
     7081 catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa
     7141 tcca
//
