LOCUS       sequence-172908-        6434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Channelrhodopsin localized to neuronal soma via a Kv2.1 targeting 
            motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6434)
  AUTHORS   Baker CA, Elyada YM, Parra A, Bolton MM
  TITLE     Cellular resolution circuit mapping with temporal-focused excitation
            of soma-targeted channelrhodopsin.
  JOURNAL   Elife. 2016 Aug 15;5. pii: e14193. doi: 10.7554/eLife.14193.
  PUBMED    27525487
REFERENCE   2  (bases 1 to 6434)
  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: "2016-08-15"; volume: "5. pii: e14193. doi"; pages: " 
            10.7554/eLife.14193"
COMMENT     Sequence Label: AAV-syn-ChR2-EYFP-Kv
FEATURES             Location/Qualifiers
     source          1..6434
                     /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        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             677..1603
                     /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      1613..1622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1619..1621,1622..1624,1625..2335)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1664..1685)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1925..1944)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2272..2293
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2561..3149
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2614..2634)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3032..3043)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3181..3657
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3380..3399)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3697..3837
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3697..3826
                     /label=AAV2 ITR
     rep_origin      3912..4367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3999..4018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4209..4230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4384..4403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4503..4525
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4563..4581)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4649..4753
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4754..4822,4823..5614)
                     /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(4972..4991)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5785..6373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6274..6293
                     /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 cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgactccgga gccaccatgg actatggcgg cgctttgtct gccgtcggac gcgaactttt
      721 gttcgttact aatcctgtgg tggtgaacgg gtccgtcctg gtccctgagg atcaatgtta
      781 ctgtgccgga tggattgaat ctcgcggcac gaacggcgct cagaccgcgt caaatgtcct
      841 gcagtggctt gcagcaggat tcagcatttt gctgctgatg ttctatgcct accaaacctg
      901 gaaatctaca tgcggctggg aggagatcta tgtgtgcgcc attgaaatgg ttaaggtgat
      961 tctcgagttc ttttttgagt ttaagaatcc ctctatgctc taccttgcca caggacaccg
     1021 ggtgcagtgg ctgcgctatg cagagtggct gctcacttgt cctgtcatcc ttatccgcct
     1081 gagcaacctc accggcctga gcaacgacta cagcaggaga accatgggac tccttgtctc
     1141 agacatcggg actatcgtgt ggggggctac cagcgccatg gcaaccggct atgttaaagt
     1201 catcttcttt tgtcttggat tgtgctatgg cgcgaacaca ttttttcacg ccgccaaagc
     1261 atatatcgag ggttatcata ctgtgccaaa gggtcggtgc cgccaggtcg tgaccggcat
     1321 ggcatggctg tttttcgtga gctggggtat gttcccaatt ctcttcattt tggggcccga
     1381 aggttttggc gtcctgagcg tctatggctc caccgtaggt cacacgatta ttgatctgat
     1441 gagtaaaaat tgttgggggt tgttgggaca ctacctgcgc gtcctgatcc acgagcacat
     1501 attgattcac ggagatatcc gcaaaaccac caaactgaac atcggcggaa cggagatcga
     1561 ggtcgagact ctcgtcgaag acgaagccga ggccggagcc gtgccagcgg ccgccaccat
     1621 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1681 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1741 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1801 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1861 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1921 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1981 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2041 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2101 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2161 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2221 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2281 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcagtc
     2341 ccagcctatt ctgaacacca aagagatggc cccccagagt aaaccccctg aggagctgga
     2401 aatgtcctca atgccctccc ctgtggcacc tcttcctgca agaacagagg gcgtgatcga
     2461 catgaggtcc atgagttcta ttgactcttt tatctcatgc gccaccgatt ttccagaggc
     2521 tacgcggttt aagtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     2581 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2641 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2701 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2761 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2821 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2881 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2941 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     3001 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3061 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3121 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     3181 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3241 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3301 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3361 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     3421 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     3481 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     3541 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     3601 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     3661 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3721 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3781 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3841 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3901 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4381 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4441 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4501 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4561 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4621 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4681 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4741 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4801 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4861 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4921 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4981 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5041 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5101 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5161 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5221 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5281 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5341 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5401 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5461 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5521 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5581 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5641 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5701 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5761 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5821 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5881 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5941 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6001 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6061 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6121 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6181 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6241 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6301 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6361 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6421 ttttgctcac atgt
//
