LOCUS       sequence-9767-ex        1662 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  3rd gen lentiviral expression of humanized ChR2 with H134R mutation 
            fused to EYFP driven by CaMKIIa promoter for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1662)
  AUTHORS   Zhang F, Wang LP, Brauner M, Liewald JF, Kay K, Watzke N, Wood PG, 
            Bamberg E, Nagel G, Gottschalk A, Deisseroth K
  TITLE     Multimodal fast optical interrogation of neural circuitry.
  JOURNAL   Nature. 2007 Apr 5. 446(7136):633-9.
  PUBMED    17410168
REFERENCE   2  (bases 1 to 1662)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2007 Apr 5. 446(7136):633-9."
COMMENT     Sequence Label: pLenti-CaMKIIa-hChR2(H134R)-EYFP-WPRE
FEATURES             Location/Qualifiers
     source          1..1662
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..927
                     /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      937..946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(943..945,946..948,949..1662)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(988..1009)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1249..1268)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1596..1617
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atggactatg gcggcgcttt gtctgccgtc ggacgcgaac ttttgttcgt tactaatcct
       61 gtggtggtga acgggtccgt cctggtccct gaggatcaat gttactgtgc cggatggatt
      121 gaatctcgcg gcacgaacgg cgctcagacc gcgtcaaatg tcctgcagtg gcttgcagca
      181 ggattcagca ttttgctgct gatgttctat gcctaccaaa cctggaaatc tacatgcggc
      241 tgggaggaga tctatgtgtg cgccattgaa atggttaagg tgattctcga gttctttttt
      301 gagtttaaga atccctctat gctctacctt gccacaggac accgggtgca gtggctgcgc
      361 tatgcagagt ggctgctcac ttgtcctgtc atccttatcc gcctgagcaa cctcaccggc
      421 ctgagcaacg actacagcag gagaaccatg ggactccttg tctcagacat cgggactatc
      481 gtgtgggggg ctaccagcgc catggcaacc ggctatgtta aagtcatctt cttttgtctt
      541 ggattgtgct atggcgcgaa cacatttttt cacgccgcca aagcatatat cgagggttat
      601 catactgtgc caaagggtcg gtgccgccag gtcgtgaccg gcatggcatg gctgtttttc
      661 gtgagctggg gtatgttccc aattctcttc attttggggc ccgaaggttt tggcgtcctg
      721 agcgtctatg gctccaccgt aggtcacacg attattgatc tgatgagtaa aaattgttgg
      781 gggttgttgg gacactacct gcgcgtcctg atccacgagc acatattgat tcacggagat
      841 atccgcaaaa ccaccaaact gaacatcggc ggaacggaga tcgaggtcga gactctcgtc
      901 gaagacgaag ccgaggccgg agccgtgcca gcggccgcca ccatggtgag caagggcgag
      961 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1021 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1081 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     1141 tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1201 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1261 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1321 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1381 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1441 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1501 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1561 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1621 gccgccggga tcactctcgg catggacgag ctgtacaagt aa
//
