LOCUS       sequence-391701-        5274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct light-gated cation channel from Chlamydomonas reinhardtii 
            that allows neuron deploarization through brief pulses of blue 
            light..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5274)
  AUTHORS   Nava S, Palma W, Wan X, Oh JY, Gharib S, Wang H, Revanna JS, Tan M, 
            Zhang M, Liu J, Chen CH, Lee JS, Perry B, Sternberg PW
  TITLE     A cGAL-UAS bipartite expression toolkit for Caenorhabditis elegans 
            sensory neurons.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Dec 19;120(51):e2221680120. doi: 
            10.1073/pnas.2221680120. Epub 2023 Dec 14.
  PUBMED    38096407
REFERENCE   2  (bases 1 to 5274)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-12-19"; volume: "120(51):e2221680120. 
            doi"; pages: " 10.1073/pnas.2221680120. Epub 2023 Dec 14"
COMMENT     Sequence Label: pHW449
FEATURES             Location/Qualifiers
     source          1..5274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..130
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    169..263
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    302..396
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     CDS             835..1761
                     /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      1771..1780
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1777..1779,1780..1782,1783..2496)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1822..1843)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2083..2102)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2430..2451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        3019..3123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3124..3192,3193..3984)
                     /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(3342..3361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4155..4743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4644..4663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4897..4914
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5031..5052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5067..5072,5073..5090,5091..5097)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5105..5121
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5110..5132
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5129..5145
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5129..5145
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcta gcccatgatt acgccaagct acgggcggag tactgtcctc cgagcggagt
       61 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact
      121 gtcctccgag cggagactcc ctagcccatg attacgccaa gctacgggcg gagtactgtc
      181 ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc
      241 cgagcggagt actgtcctcc gagcggagac tccctagccc atgattacgc caagctacgg
      301 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg
      361 gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gactccctag gggcggccgc
      421 actgactggg ccggccaaaa atgcataagg ttttgctggc actgttcttt atctttctgg
      481 caccagcatc cgcactggca gtctccgaac cggcctgcag gtcgattttt tgcaaattac
      541 gagcgttgta gggggcggag cgataggtcc tataggtttt ggtatatcat cattcattca
      601 ttcattggta cattcattta cccaccttcc tctttctgag cttctctgga gttctgtgct
      661 tcctttttcc cttatcttta tactgtaatt tttaactttc aggcattgat tggcgcgcct
      721 ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg atactaacat
      781 aacatagaac attttcagga ggacccttgg ctagcgtcga cggtaccggt aaaaatggac
      841 tatggcggcg ctttgtctgc cgtcggacgc gaacttttgt tcgttactaa tcctgtggtg
      901 gtgaacgggt ccgtcctggt ccctgaggat caatgttact gtgccggatg gattgaatct
      961 cgcggcacga acggcgctca gaccgcgtca aatgtcctgc agtggcttgc agcaggattc
     1021 agcattttgc tgctgatgtt ctatgcctac caaacctgga aatctacatg cggctgggag
     1081 gagatctatg tgtgcgccat tgaaatggtt aaggtgattc tcgagttctt ttttgagttt
     1141 aagaatccct ctatgctcta ccttgccaca ggacaccggg tgcagtggct gcgctatgca
     1201 gagtggctgc tcacttgtcc tgtcatcctt atccgcctga gcaacctcac cggcctgagc
     1261 aacgactaca gcaggagaac catgggactc cttgtctcag acatcgggac tatcgtgtgg
     1321 ggggctacca gcgccatggc aaccggctat gttaaagtca tcttcttttg tcttggattg
     1381 tgctatggcg cgaacacatt ttttcacgcc gccaaagcat atatcgaggg ttatcatact
     1441 gtgccaaagg gtcggtgccg ccaggtcgtg accggcatgg catggctgtt tttcgtgagc
     1501 tggggtatgt tcccaattct cttcattttg gggcccgaag gttttggcgt cctgagcgtc
     1561 tatggctcca ccgtaggtca cacgattatt gatctgatga gtaaaaattg ttgggggttg
     1621 ttgggacact acctgcgcgt cctgatccac gagcacatat tgattcacgg agatatccgc
     1681 aaaaccacca aactgaacat cggcggaacg gagatcgagg tcgagactct cgtcgaagac
     1741 gaagccgagg ccggagccgt gccagcggcc gccaccatgg tgagcaaggg cgaggagctg
     1801 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1861 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1921 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
     1981 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2041 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2101 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2161 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2221 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2281 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2341 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     2401 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2461 gggatcactc tcggcatgga cgagctgtac aagtaagaat tcgctagccg gccatacaag
     2521 taatccggat gatcgacgcc aacgtcgttg aattttcaaa ttttaaatac tgaatatttg
     2581 ttttttttcc tattatttat ttattctctt tgtgtttttt ttcttgcttt ctaaaaaatt
     2641 aattcaatcc aaatctaaac attttttttt ctctttccgt ctcccaattc gtattccgct
     2701 cctctcatct gaacacaatg tgcaagttta tttatcttct cgctttcatt tcattaggac
     2761 gtggggggaa ttggtggaag ggggaaacac acaaaaggat gatggaaatg aaataaggac
     2821 acacaatatg caacaacatt caattcagaa atatggagga aggtttaaaa gaaaacataa
     2881 aaatatatag aggaggaagg aaaactagta aaaaataagc aaagaaatta ggcgaacgat
     2941 gagaattgtc ctcgcttggg cccaaaaggc cttttttttt tggcgccgac gtcaggtggc
     3001 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3061 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3121 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3181 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3241 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3301 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3361 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3421 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3481 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3541 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3601 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3661 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3721 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3781 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3841 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3901 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3961 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4021 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4081 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4141 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4201 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4261 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4321 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4381 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4441 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4501 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4561 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4621 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4681 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4741 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4801 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4861 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4921 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4981 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5041 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5101 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     5161 ctgtaagttt aaacatgatc ttactaacta actattctca tttaaatttt cagagcttaa
     5221 aaatggctga aatcactcac aacgatggat acgctaacaa cttggaaatg aaat
//
