LOCUS       sequence-334105-        7248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dre-Dependent ChR2-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7248)
  AUTHORS   Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone
            J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H,
            Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, 
            Deisseroth K
  TITLE     Targeting cells with single vectors using multiple-feature Boolean 
            logic.
  JOURNAL   Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 
            2014 Jun 8.
  PUBMED    24908100
REFERENCE   2  (bases 1 to 7248)
  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.2996"; journalName: "Nat Methods"; date: "2014-07"; 
            volume: "11"; issue: "7"; pages: "763-72"
COMMENT     Sequence Label: pAAV-Ef1a-dDIO hChR2(H134R)-EYFP
FEATURES             Location/Qualifiers
     source          1..7248
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1440..1471
                     /label=rox
                     /bound_moiety="Dre recombinase"
     CDS             complement(join(1565..2278,2279..2281,2282..2284))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1610..1631)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1959..1978
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2218..2239
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2281..2290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2300..3226)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      3223..3232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3237..3268
                     /label=rox
                     /bound_moiety="Dre recombinase"
     misc_feature    3375..3963
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3428..3448)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3846..3857)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3995..4471
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4194..4213)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4511..4651
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4511..4640
                     /label=AAV2 ITR
     rep_origin      4726..5181
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4813..4832)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5023..5044
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5198..5217)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5317..5339
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5377..5395)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5463..5567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5568..5636,5637..6428)
                     /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(5786..5805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6599..7187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7088..7107
                     /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 gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggat
     1441 aactttaaat aatgccaatt atttaaagtt agcagaatgg tagctggatt gtagctgcta
     1501 ttagcaatat gaaacctctt ataactttaa ataatgtcga ttatttaaag ttaggcgcgc
     1561 cctattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     1621 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tggtagctca
     1681 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     1741 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     1801 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     1861 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     1921 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     1981 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     2041 gctgcttcat gtggtcgggg tagcgggcga agcactgcag gccgtagccg aaggtggtca
     2101 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     2161 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     2221 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     2281 ccatggtggc ggccgctggc acggctccgg cctcggcttc gtcttcgacg agagtctcga
     2341 cctcgatctc cgttccgccg atgttcagtt tggtggtttt gcggatatct ccgtgaatca
     2401 atatgtgctc gtggatcagg acgcgcaggt agtgtcccaa caacccccaa caatttttac
     2461 tcatcagatc aataatcgtg tgacctacgg tggagccata gacgctcagg acgccaaaac
     2521 cttcgggccc caaaatgaag agaattggga acatacccca gctcacgaaa aacagccatg
     2581 ccatgccggt cacgacctgg cggcaccgac cctttggcac agtatgataa ccctcgatat
     2641 atgctttggc ggcgtgaaaa aatgtgttcg cgccatagca caatccaaga caaaagaaga
     2701 tgactttaac atagccggtt gccatggcgc tggtagcccc ccacacgata gtcccgatgt
     2761 ctgagacaag gagtcccatg gttctcctgc tgtagtcgtt gctcaggccg gtgaggttgc
     2821 tcaggcggat aaggatgaca ggacaagtga gcagccactc tgcatagcgc agccactgca
     2881 cccggtgtcc tgtggcaagg tagagcatag agggattctt aaactcaaaa aagaactcga
     2941 gaatcacctt aaccatttca atggcgcaca catagatctc ctcccagccg catgtagatt
     3001 tccaggtttg gtaggcatag aacatcagca gcaaaatgct gaatcctgct gcaagccact
     3061 gcaggacatt tgacgcggtc tgagcgccgt tcgtgccgcg agattcaatc catccggcac
     3121 agtaacattg atcctcaggg accaggacgg acccgttcac caccacagga ttagtaacga
     3181 acaaaagttc gcgtccgacg gcagacaaag cgccgccata gtccatggtg gctagctaac
     3241 tttaaataat tggcattatt taaagttatt gccttaaccc agaaattatc actgttattc
     3301 tttagaatgg tgcaaagata actttaaata atcgacatta tttaaagtta gaattcgata
     3361 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3421 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3481 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3541 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3601 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3661 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3721 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3781 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3841 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3901 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3961 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     4021 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     4081 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     4141 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     4201 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     4261 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     4321 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     4381 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     4441 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     4501 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     4561 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     4621 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     4681 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     4741 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4801 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4861 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4921 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4981 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5041 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     5101 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5161 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     5221 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5281 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5341 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5401 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5461 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5521 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5581 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5641 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5701 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5761 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5821 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5881 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5941 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6001 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6061 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6121 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6181 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6241 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6301 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6361 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6421 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6481 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6541 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6601 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6661 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6721 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6781 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6841 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6901 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6961 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7021 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7081 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7141 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7201 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
