LOCUS       sequence-391691-        5336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Step-Waveform Inhibitory Channelrhodopsin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5336)
  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 5336)
  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: pJR034
FEATURES             Location/Qualifiers
     source          1..5336
                     /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             join(1834..1836,1837..2550)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1876..1897)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2137..2156)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2484..2505
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        3081..3185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3186..3254,3255..4046)
                     /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(3404..3423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4217..4805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4706..4725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4959..4976
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5093..5114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5129..5134,5135..5152,5153..5159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5167..5183
                     /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     5172..5194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5191..5207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5191..5207
                     /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 ctcttccaaa cctcatacac tcttgagaac
      901 aatggttctg ttatttgcat cccgaacaac ggccagtgct tctgcttggc ttggcttaaa
      961 tccaacggaa caaatgccga gaagttggct gccaacattc tgcagtggat ttcttttgcg
     1021 ctttcagcgc tctgcctgat gttctacggc taccagacct ggaagtctac ttgcggctgg
     1081 gaggagattt acgtggccac gatctcgatg atcaagttca tcatcgagta tttccatagc
     1141 tttgacgaac ctgcggtgat ctactcatcc aacggcaaca agaccaagtg gcttcgttac
     1201 gcgagctggc tgctgaccgc ccctgtcatt cttatccgtc tgagcaacct tacgggtctg
     1261 gcgaacgact ataacaagcg taccatgggt ctgctggtgt cagatatcgg cacgatcgtg
     1321 tggggcacca cggccgcgct gtccaaggga tacgtccgtg tcattttctt cctgatgggc
     1381 ctgtgctacg gcatctacac attcttcaac gcagccaagg tctacattga ggcgtaccac
     1441 accgtgccga agggccggtg tcgccaggtg gtgactggca tggcttggct cttcttcgta
     1501 tcatggggta tgttccccat cctgttcatc ctcggccccg agggcttcgg cgtcctgagc
     1561 aagtacggct ccaacgtcgg ccacaccatc attgacctga tgtcgaagca gtgctggggt
     1621 ctgctcggcc actacctgcg cgtgctgatc cacgagcata tcctcatcca cggcgacatt
     1681 cgcaagacca ccaaattgaa cattggtggc actgagattg aggtcgagac gctggtggag
     1741 gacgaggccg aggctggcgc ggtcgcggcc gccaagagca ggatcaccag cgagggcgag
     1801 tacatccccc tggaccagat cgacatcaac gtggtgagca agggcgagga gctgttcacc
     1861 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1921 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1981 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag
     2041 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2101 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2161 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2221 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2281 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2341 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2401 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     2461 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2521 actctcggca tggacgagct gtacaagtaa cattcgtaga attcgctagc cggccataca
     2581 agtaatccgg atgatcgacg ccaacgtcgt tgaattttca aattttaaat actgaatatt
     2641 tgtttttttt cctattattt atttattctc tttgtgtttt ttttcttgct ttctaaaaaa
     2701 ttaattcaat ccaaatctaa acattttttt ttctctttcc gtctcccaat tcgtattccg
     2761 ctcctctcat ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca tttcattagg
     2821 acgtgggggg aattggtgga agggggaaac acacaaaagg atgatggaaa tgaaataagg
     2881 acacacaata tgcaacaaca ttcaattcag aaatatggag gaaggtttaa aagaaaacat
     2941 aaaaatatat agaggaggaa ggaaaactag taaaaaataa gcaaagaaat taggcgaacg
     3001 atgagaattg tcctcgcttg ggcccaaaag gccttttttt tttggcgccg acgtcaggtg
     3061 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3121 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3181 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3241 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3301 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3361 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3421 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3481 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3541 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3601 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3661 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3721 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3781 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3841 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3901 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3961 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4021 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4081 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4141 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4201 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4261 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4321 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4381 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4441 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4501 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4561 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4621 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4681 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4741 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4801 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4861 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4921 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4981 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5041 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5101 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5161 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     5221 agctgtaagt ttaaacatga tcttactaac taactattct catttaaatt ttcagagctt
     5281 aaaaatggct gaaatcactc acaacgatgg atacgctaac aacttggaaa tgaaat
//
