LOCUS       sequence-391889-        5285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stabilized step function Opsins ChR2 variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5285)
  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 5285)
  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: pJR032
FEATURES             Location/Qualifiers
     source          1..5285
                     /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             838..1764
                     /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
                     PSMLYLATGHRVQWLRYAEWLLTSPVILIHLSNLTGLSNDYSRRTMGLLVSAIGTIVCG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      1774..1783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1780..1782,1783..1785,1786..2499)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1825..1846)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2086..2105)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2433..2454
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        3030..3134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3135..3203,3204..3995)
                     /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(3353..3372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4166..4754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4655..4674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4908..4925
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5042..5063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5078..5083,5084..5101,5102..5108)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5116..5132
                     /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     5121..5143
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5140..5156
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5140..5156
                     /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 agaaaaaatg
      841 gactatggcg gcgctttgtc tgccgtcgga cgcgaacttt tgttcgttac taatcctgtg
      901 gtggtgaacg ggtccgtcct ggtccctgag gatcaatgtt actgtgccgg atggattgaa
      961 tctcgcggca cgaacggcgc tcagaccgcg tcaaatgtcc tgcagtggct tgcagcagga
     1021 ttcagcattt tgctgctgat gttctatgcc taccaaacct ggaaatctac atgcggctgg
     1081 gaggagatct atgtgtgcgc cattgaaatg gttaaggtga ttctcgagtt cttttttgag
     1141 tttaagaatc cctctatgct ctaccttgcc acaggacacc gggtgcagtg gctgcgctat
     1201 gcagagtggc tgctcacttc tcctgtcatc cttatccacc tgagcaacct caccggcctg
     1261 agcaacgact acagcaggag aaccatggga ctccttgtct cagccatcgg gactatcgtg
     1321 tgtggggcta ccagcgccat ggcaaccggc tatgttaaag tcatcttctt ttgtcttgga
     1381 ttgtgctatg gcgcgaacac attttttcac gccgccaaag catatatcga gggttatcat
     1441 actgtgccaa agggtcggtg ccgccaggtc gtgaccggca tggcatggct gtttttcgtg
     1501 agctggggta tgttcccaat tctcttcatt ttggggcccg aaggttttgg cgtcctgagc
     1561 gtctatggct ccaccgtagg tcacacgatt attgatctga tgagtaaaaa ttgttggggg
     1621 ttgttgggac actacctgcg cgtcctgatc cacgagcaca tattgattca cggagatatc
     1681 cgcaaaacca ccaaactgaa catcggcgga acggagatcg aggtcgagac tctcgtcgaa
     1741 gacgaagccg aggccggagc cgtgccagcg gccgccacca tggtgagcaa gggcgaggag
     1801 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1861 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1921 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     1981 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2041 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2101 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2161 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2221 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2281 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2341 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     2401 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2461 gccgggatca ctctcggcat ggacgagctg tacaagtaac attcgtagaa ttcgctagcc
     2521 ggccatacaa gtaatccgga tgatcgacgc caacgtcgtt gaattttcaa attttaaata
     2581 ctgaatattt gttttttttc ctattattta tttattctct ttgtgttttt tttcttgctt
     2641 tctaaaaaat taattcaatc caaatctaaa catttttttt tctctttccg tctcccaatt
     2701 cgtattccgc tcctctcatc tgaacacaat gtgcaagttt atttatcttc tcgctttcat
     2761 ttcattagga cgtgggggga attggtggaa gggggaaaca cacaaaagga tgatggaaat
     2821 gaaataagga cacacaatat gcaacaacat tcaattcaga aatatggagg aaggtttaaa
     2881 agaaaacata aaaatatata gaggaggaag gaaaactagt aaaaaataag caaagaaatt
     2941 aggcgaacga tgagaattgt cctcgcttgg gcccaaaagg cctttttttt ttggcgccga
     3001 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3061 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3301 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3361 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4801 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4861 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4921 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4981 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5041 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5101 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5161 attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc atttaaattt
     5221 tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca acttggaaat
     5281 gaaat
//
