LOCUS       sequence-215859-        7148 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 7148)
  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 7148)
  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..7148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1420
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1411
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1368..1388
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1451..1484)
                     /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)."
     protein_bind    complement(1535..1568)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1586..2287)
                     /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(1669..1692)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1867..1886)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2136..2154
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(2288..2344)
                     /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"
     regulatory      3406..3415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3425..3458
                     /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)."
     protein_bind    3509..3542
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3567..4155
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3620..3640)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4038..4049)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4188..4299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4411..4551
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4411..4540
                     /label=AAV2 ITR
     rep_origin      4626..5081
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4713..4732)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4923..4944
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5098..5117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5217..5239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5277..5295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5363..5467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5468..5536,5537..6328)
                     /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(5686..5705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6499..7087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6988..7007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
      601 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
      661 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      721 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      781 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
      841 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      901 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      961 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1021 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     1081 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     1141 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1201 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1261 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1321 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1381 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagagt
     1441 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
     1501 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
     1561 gaagttatgg cgcgccgtcg acttacttgt acagctcgtc catgccgtac aggaacaggt
     1621 ggtggcggcc ctcggagcgc tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg
     1681 tgatgtccag cttggtgtcc acgtagtagt agccgggcag ttgcacgggc ttcttggcca
     1741 tgtagatggt cttgaactcc accaggtagt ggccgccgtc cttcagcttc agggcctggt
     1801 ggatctcgcc cttcagcacg ccgtcgcggg ggtacaggcg ctcggtggag gcctcccagc
     1861 ccatggtctt cttctgcatt acggggccgt cgggggggaa gttggtgccg cgcatcttca
     1921 ccttgtagat cagcgtgccg tcctgcaggg aggagtcctg ggtcacggtc accagaccgc
     1981 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcttgt
     2041 aatcggggat gtcggcgggg tgcttcacgt acgccttgga gccgtacatg aactgggggg
     2101 acaggatgtc ccaggcgaag ggcagggggc cgcccttggt caccttcagc ttggcggtct
     2161 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     2221 tggagccctc catgcgcacc ttgaagcgca tgaactcttt gatgacctcc tcgcccttgc
     2281 tcaccattgg cccgggattc tcttcgacat cccctgcttg tttcaacagg gagaagttag
     2341 tggctccgct tccgttaacg ctgctctcgt acttatcttc ttcttccgcc accagggttt
     2401 ccacttccat ttcctggccc gcaatggtaa ttttctgttt tttgcgaata tcgccataca
     2461 gcagaatatg ttcatgaatt ttcacgcgca gatagttgcc cagcacgccc cacatgttct
     2521 tcgcaatcag atccagaatg ctatggccaa tcgcgctgcc atacgggcta atatggccga
     2581 agccctcggg gcccagcagg aacagcacgg ggaacatgcc ccagctcacg aagaacagcc
     2641 aggccatggc tctcaccagc tgtctgcaca ggcctttcgg cacggtatga aacgcttcaa
     2701 tatacacttt ggccgcatga aaataggtat acatgccata gctcaggcta atcagaaaaa
     2761 acagaatttt ggtccagccg gtgcacatcg cgctggtcgc gccccacaca atgcagccca
     2821 cgtcgctcac cagcaggccc atggtgcgtt tgctataatc atctttcagg ccggtcaggt
     2881 tgctcagatg aatcagaatc acggggcagg tcagcagcca ttcgccatag cgcatccaca
     2941 ccacgccgtt gccgctgctc agccacaggg tcgccgggct atcaaattca tgaaacgctt
     3001 caataatgct tttcatcatt tcaatcagcg ccacatacac ttcttcccag ccgcaggtcg
     3061 cgcgccacgc ctgatacgca taccagccca ggcacgccac gctcagcgca aacaccaccc
     3121 actgcaggat gttggcggcc agcttctcgg cgttggtgcc gttactcttc agccaggcca
     3181 ggcagaagca ctggccgttg ttggggatgc agatcacgct gccgttgttc tccagggtgt
     3241 agctggtctg gaacagcatt ctctcgtggg ctctgtggaa cacgtagtcg gggccgtcct
     3301 gggtggccac gggcacggtg gcgtcgctgc cggtgctggc gccggcgctg ccggcggcca
     3361 gggccacggc cagggccagg gccagcagcc agggtcttct gctcaccatg gtggcgccgc
     3421 tagcataact tcgtataaag tatcctatac gaagttattt gccttaaccc agaaattatc
     3481 actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct atacgaagtt
     3541 atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3601 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3661 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3721 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3781 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3841 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3901 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3961 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     4021 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     4081 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     4141 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctcgactg tgccttctag
     4201 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4261 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4321 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4381 caggcatgca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     4441 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4501 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     4561 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     4621 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4681 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4741 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4801 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     4861 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4921 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     4981 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5041 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5101 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5161 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5221 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5281 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5341 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5401 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5461 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5521 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5581 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5641 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5701 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5761 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5821 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5881 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5941 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6001 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6061 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6121 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6181 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6241 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6301 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6361 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6421 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6481 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6541 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6601 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     6661 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6721 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6781 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6841 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6901 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6961 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7021 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7081 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7141 tcacatgt
//
