LOCUS       sequence-203174-        6933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Channelrhodopsin localized to neuronal soma with simultaneous 
            nuclear labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6933)
  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 6933)
  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-Kv-P2A-H2b-mRuby2
FEATURES             Location/Qualifiers
     source          1..6933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      229..238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             235..1161
                     /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"
     CDS             1381..1437
                     /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             1444..1821
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1885..1894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1891..2601
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    2629..3217
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2682..2702)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3100..3111)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3249..3725
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3448..3467)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3765..3905
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3765..3894
                     /label=AAV2 ITR
     rep_origin      3980..4435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4067..4086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4277..4298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4452..4471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4571..4593
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4631..4649)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4822..4890,4891..5682)
                     /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(5040..5059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5853..6441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6342..6361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6503..6632
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(6677..6933,1..191)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
       61 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      121 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      181 tgagagcgca gtcgagaagg taccggatcc tctagagtcg actccggagc caccatggac
      241 tatggcggcg ctttgtctgc cgtcggacgc gaacttttgt tcgttactaa tcctgtggtg
      301 gtgaacgggt ccgtcctggt ccctgaggat caatgttact gtgccggatg gattgaatct
      361 cgcggcacga acggcgctca gaccgcgtca aatgtcctgc agtggcttgc agcaggattc
      421 agcattttgc tgctgatgtt ctatgcctac caaacctgga aatctacatg cggctgggag
      481 gagatctatg tgtgcgccat tgaaatggtt aaggtgattc tcgagttctt ttttgagttt
      541 aagaatccct ctatgctcta ccttgccaca ggacaccggg tgcagtggct gcgctatgca
      601 gagtggctgc tcacttgtcc tgtcatcctt atccgcctga gcaacctcac cggcctgagc
      661 aacgactaca gcaggagaac catgggactc cttgtctcag acatcgggac tatcgtgtgg
      721 ggggctacca gcgccatggc aaccggctat gttaaagtca tcttcttttg tcttggattg
      781 tgctatggcg cgaacacatt ttttcacgcc gccaaagcat atatcgaggg ttatcatact
      841 gtgccaaagg gtcggtgccg ccaggtcgtg accggcatgg catggctgtt tttcgtgagc
      901 tggggtatgt tcccaattct cttcattttg gggcccgaag gttttggcgt cctgagcgtc
      961 tatggctcca ccgtaggtca cacgattatt gatctgatga gtaaaaattg ttgggggttg
     1021 ttgggacact acctgcgcgt cctgatccac gagcacatat tgattcacgg agatatccgc
     1081 aaaaccacca aactgaacat cggcggaacg gagatcgagg tcgagactct cgtcgaagac
     1141 gaagccgagg ccggagccgt gccagcggcc gccacccagt cccagcctat tctgaacacc
     1201 aaagagatgg ccccccagag taaaccccct gaggagctgg aaatgtcctc aatgccctcc
     1261 cctgtggcac ctcttcctgc aagaacagag ggcgtgatcg acatgaggtc catgagttct
     1321 attgactctt ttatctcatg cgccaccgat tttccagagg ctacgcggtt tggatccgga
     1381 gccacgaact tctctctgtt aaagcaagca ggagacgtgg aagaaaaccc cggtcctggt
     1441 tctatgccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa gaaggcggtg
     1501 actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga gagctattcc
     1561 atctatgtgt acaaggttct gaagcaggtc caccctgaca ccggcatttc gtccaaggcc
     1621 atgggcatca tgaattcgtt tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc
     1681 cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca gacggccgtg
     1741 cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac taaggccatc
     1801 accaagtaca ccagcgctaa ggatccaccg gtcgccacca tgctcgtcga agacgaagcc
     1861 gaggccggag ccgtgccagc ggccgccacc atggtgtcta agggcgaaga gctgatcaag
     1921 gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca attcaaatgc
     1981 acaggtgaag gagaaggcaa tccgtacatg ggaactcaaa ccatgaggat caaagtcatc
     2041 gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat gtatggcagc
     2101 cgtactttta tcaagtaccc gaaaggcatt cctgatttct ttaaacagtc ctttcctgag
     2161 ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac cgtcatgcag
     2221 gacaccagcc ttgaggatgg ctgtctcgtt taccacgtcc aagtcagagg ggtaaacttt
     2281 ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa tacagagatg
     2341 atgtatccag cagatggtgg tctgagggga tacactcata tggcactgaa agttgatggt
     2401 ggtggccatc tgtcttgctc tttcgtaaca acttacaggt caaaaaagac cgtcgggaac
     2461 atcaagatgc ccggtatcca tgccgttgat caccgcctgg aaaggttaga ggaaagtgac
     2521 aatgaaatgt tcgtagtaca acgcgaacac gcagttgcca agttcgccgg gcttggtggt
     2581 ggaatggacg agctgtacaa gtaagaattc gatatcaagc ttatcgataa tcaacctctg
     2641 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2701 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2761 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2821 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2881 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2941 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3001 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3061 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3121 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3181 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3241 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3301 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3361 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3421 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3481 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3541 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3601 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3661 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3721 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3781 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3841 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3901 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3961 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4021 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4081 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4141 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4201 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4261 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4321 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4381 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4441 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4501 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4561 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4621 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4681 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4861 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4921 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4981 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5041 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5101 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5161 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5221 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5281 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5341 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5401 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5461 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5521 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5581 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5641 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5701 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5761 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5821 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5881 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5941 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6001 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6061 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6481 tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     6541 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     6601 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca
     6661 gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg
     6721 cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg
     6781 cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc
     6841 cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc
     6901 tcagcactga aggcgcgctg acgtcactcg ccg
//
