LOCUS       sequence-466494-        9697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the optogenetic tool BiPOLES in cholinergic neurons of C. 
            elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9697)
  AUTHORS   Bergs ACF, Liewald JF, Rodriguez-Rozada S, Liu Q, Wirt C, Bessel A, 
            Zeitzschel N, Durmaz H, Nozownik A, Dill H, Jospin M, Vierock J, 
            Bargmann CI, Hegemann P, Wiegert JS, Gottschalk A
  TITLE     All-optical closed-loop voltage clamp for precise control of muscles
            and neurons in live animals.
  JOURNAL   Nat Commun. 2023 Apr 6;14(1):1939. doi: 10.1038/s41467-023-37622-6.
  PUBMED    37024493
REFERENCE   2  (bases 1 to 9697)
  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: "Nat 
            Commun."; date: "2023-04-6"; volume: "14(1):1939. doi"; pages: " 
            10.1038/s41467-023-37622-6"
COMMENT     Sequence Label: pAB26 
            (punc-17::GtACR2::mCerulean::beta-HK::Chrimson)
FEATURES             Location/Qualifiers
     source          1..9697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4300..4321)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4561..4580)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4908..4929
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5329..6375
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=Chrimson
                     /translation="ELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTEWW
                     CDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSIAI
                     ALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRYFE
                     WLLSCPVILIKLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIVSC
                     IYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLKLS
                     PYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEEFV
                     EEEDEDTV"
     primer_bind     complement(7222..7244)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7344..7363
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7441..7459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7527..7631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7632..7700,7701..8492)
                     /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(7850..7869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8663..9251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9152..9171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9405..9422
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9539..9560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9575..9580,9581..9598,9599..9605)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9613..9629
                     /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     9618..9640
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9637..9653
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9637..9653
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      9683..9692
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ggattacacc aatcatttct cccctccgaa atgtgtagtc gtcatgacaa agtggtgaca
       61 ctggattacg gtaggctccg cccacttctt ggaaaagtga cgtgggctcg gtgagcccta
      121 aagcttttgg ttttcacaat tttctggttt ttgggatttt tgcgggaatt ttgagaggag
      181 aaagttggga gagaagatat tggaatggtc cccggggagc tgtacattta tacatctaga
      241 ctgaagctgg ggtattgcaa aaagagaaaa gcctatcact ttctttccat cctgcatact
      301 tgccaaaaag ttttggctga gcttattttt aaaattggaa ttttttggaa ttttcagaaa
      361 agaaaaagtt ttttcttttt ttttaattta aaaaattttt gctatttatt gcatggtact
      421 acagaaaaac ttgaatgaaa taaaaaattt ggaaaattca agaaaacttt ttttttcata
      481 gtttatttat tcgagtttta atgtggcttt tattttgttc ggtttttgtt ttaaaaagat
      541 tttatacaaa atttcaagaa tttacaattt ttccgttgta aaaaatccaa aaaaaaatcc
      601 aaaaaaatag ttcagacctt tttttaaatt ttaaatttaa aaattcaata ccaaaaaatt
      661 aaaaaaaatt agtttttccg agaataaagg aaaaatgttg aaaaaatatt atatataaaa
      721 tttccagaaa atgtgaaaac tccaaaaaac tgcagacttt tccccaaact agctattttt
      781 tgaattcatt acctttaaag gtggagtacc gaaatttaag actttgcttt tttagaccca
      841 aaatggtcca aaactaccaa atttcgtaat gagacgttct gaaaattttt caaaaaaatg
      901 ttatgggagc tcaaagtttt tgaaaaaatg gtatattttt agctaatttt aaccacttct
      961 tggcgtcgca gcggttggaa ctcaatttta attaaattat catccttcaa tgtccttcaa
     1021 tctttgcaat ttcaatcgac catacgcgtt cagagcattt ttttttttcg tttttcctcc
     1081 cgtccagcac atatttaaat attctctctc cccctcccct ggttcctaaa aacttgggaa
     1141 aggggaattt cgaaggaggt ggaagcttct tcttctcccc gagcacacac acacatcctc
     1201 tcttcaaaat ctttaagaaa tagtgcgtgt gaatgactag acgctgaagc taacttggtt
     1261 tccgcgaaca atcccaatca gtgcctgtgc cgaggcttcg gagaagaaga gattgacgtc
     1321 gtctttttct ttttttgtga aacagaataa aaaagaatac gaaatggact ttttcacaca
     1381 ttttcgttac ttttccagaa aaaaacgaag ctctagccat ggaagtttga attgaaaacc
     1441 taaaccccga tgtgtattca tttgagctta agcctaagcc taattctacg cctaagcata
     1501 agcctaagcc tatgttcaag cctaagccaa attgtagaac gttaccaaaa gatcaaacct
     1561 atttttccta cagaaaaggt ctaatgataa ctgttaagcg agagacaagg agagatatat
     1621 atcaaaaact agcccaaact gtgattaaca tctattaact gacctcaaaa aaaaaaatag
     1681 tagagattaa atactgctca aaaagtttct tggaaaagta aattgttagg tttctggggc
     1741 ctgtaaaact ggaccgagtt ttgaagaatt tttttaactt ttcaaagttt ggcgtcactg
     1801 aaagctttat attgtacatg ttccaccaat tttcttttgt atccgttttt tctgtctcaa
     1861 aacggagctc gagcccatga cgtgtggata ccgtgagaat ccgcccaatt ccttcacatc
     1921 gatctcatat ttctttccca gtttatattg aattcccgtt gaattgcggc tttgcagggg
     1981 agaagatgtg cgtctttata tttttttttc tatccttacc cgccgtgaat ccggctcgaa
     2041 ttgagccaat ttttatgaat agtagagcta attgacttcg tctccctaga attgatgagg
     2101 taaagctcgt ggggagatat ggagattttt cgattttttt tccagtaatt ttcagttcaa
     2161 aatttgagct tttgagcagg agttttgaga tttgtgccaa aatttcctat tattttttca
     2221 aatatttttg cttttgtaaa aaaggtgtag tagaagtttt gtagtgccac tatttttttt
     2281 ttgaaaatta tgttttggcg gaaagttctt taaaaaagtt aatcaaaatt tttggcgctt
     2341 tagtcaaatt aattttctgg caaaaatatt agtaattttg gcgtgagttc aattttccca
     2401 ttcttaaaat ttcacacaat taagaatttt aagatttggg tttttttttc aacaaaaaat
     2461 caaatattta catgaaaatt cggcgggaaa taaaaaaaaa tattccgatt caatgtgtga
     2521 ataacttttt gcggggaatt caaatttgca ttatttaaac tttttttaag ctcgaaaaat
     2581 tgaaattctt aaataaataa ataaaatttt cctcgaattt tgaatgtttc gattttttag
     2641 cgttattttt tcaaaatttc cactaaaact ctagttctaa agttctaaaa ctttaaaaac
     2701 ttcctaatgt tgtggtcttg tgtttttttg gcagcgattt ccaaatttct gaaattccaa
     2761 caaaaaaacc actacgtcag tttgcatatc caaccccccc cccccccccc tctgagccaa
     2821 tcaatcatgt gcattttgaa ttggcgccaa ggcaatcttc gtaaaaaata acaatagagt
     2881 catccctcag aacagaaaag aaacccgccc gggggctcaa atacacatgt catttttctt
     2941 ttttcgcctg atttttagcc taaattagtg tttagcaatt ttttaaggta actcggtgga
     3001 gtcatttgaa ctttttaaac taaaatttca gaaaaaattc tttttttttg tcaaaaaata
     3061 attttcccgc cggaattttt caaaaaaata atttttattt tatttttcaa gttccttgtt
     3121 tcctctttat ttttttattt tgcccccaaa agttttacac taaaatattt aattttcagg
     3181 taattcatcg aaagtctttc tattttccgc atctcttgtt caagggattg gccaaaggac
     3241 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
     3301 atggcatcac aggtcgtcta tggggaatgg gcatcaactc acaccgagtg ctacaatatg
     3361 tccagaatcg actccacctt cgtctcactg ctgcagctgg tgtgggccgt ggtctcaggc
     3421 tgccagacaa tcttcatgat tagcagggct cccaaggtcc cttgggagtc cgtgtacctg
     3481 ccctttgtcg aatctatcac ttatgcactg gccagtactg gcaacggaac cctgcagatg
     3541 agggacggca gattctttcc ttggagccga atggcctcct ggctgtgcac ctgtccaatc
     3601 atgctgggac agatttctaa catggctctg gtgaagtaca aaagtatccc actgaatccc
     3661 attgcacagg ccgcttccat cattagagtg gtcatgggaa tcaccgctac aatttctccc
     3721 gcagagtata tgaagtggct gttctttttc tttggggcca cctgcctggt gttcgagtac
     3781 agtgtggtct tcacaatctt tcaagtgggg ctgtatggct ttgaatctgt cgggacacct
     3841 ctggcacaga aggtggtggt gaggatcaag atgctgcgcc tgatcttctt tattgcttgg
     3901 actatgttcc caatcgtgtg gctgatttca cccaccggag tctgcgtgat ccacgagaac
     3961 gtgagcgcca ttctgtacct gctggctgac ggcctgtgca aaaatacata tggagtgatc
     4021 ctgtggagca ctgcctgggg agtgctggaa gggaagtggg accccgcatg tctgccaggg
     4081 caggagaaac ccgaagccga cgatcctttt ggcctgaatc acgagaagaa tgcccctcct
     4141 aacgacgaag tcaatatcag aatgtttgga aggggacgta cggcggccaa gagcaggatc
     4201 accagcgagg gcgagtacat ccccctggac cagatcgaca tcaacgtggc ggtacccgtg
     4261 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     4321 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     4381 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     4441 accaccctga gctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     4501 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     4561 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     4621 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     4681 gagtacaacg ccatccacgg caacgtctat atcaccgccg acaagcagaa gaacggcatc
     4741 aaggccaact tcggcctcaa ctgcaacatc gaggacggca gcgtgcagct cgccgaccac
     4801 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     4861 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     4921 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtatac cggtgcagcc
     4981 ctgcaggaga agaagtcatg cagccagcgc atggccgaat tccggcaata ctgttggaac
     5041 ccggacactg ggcagatgct gggccgcacc ccagcccggt gggtgtggat cagcctgtac
     5101 tatgcagctt tctacgtggt catgactggg ctctttgcct tgtgcatcta tgtgctgatg
     5161 cagaccattg atccctacac ccccgactac caggaccagt taaagtcacc gggggtaacc
     5221 ttgagaccgg atgtgtatgg ggaaagaggg ctgcagattt cctacaacat ctctgaaaac
     5281 agctctagac aggcccagat caccggacgt ccggagggag ctgtacaaga gctgatcagc
     5341 agcgccacca gatctctgtt tgccgccgga ggcatcaacc cttggcctaa cccctaccac
     5401 cacgaggaca tgggctgtgg aggaatgaca cctacaggcg agtgcttcag caccgagtgg
     5461 tggtgtgacc cttcttacgg actgagcgac gccggatacg gatattgctt cgtggaggcc
     5521 acaggcggct acctggtcgt gggagtggag aagaagcagg cttggctgca cagcagaggc
     5581 acaccaggag aaaagatcgg cgcccaggtc tgccagtgga ttgctttcag catcgccatc
     5641 gccctgctga cattctacgg cttcagcgct tggaaggcca cttgcggttg ggaggaggtc
     5701 tacgtctgtt gcgtcgaggt gctgttcgtg accctggaga tcttcaagga gttcagcagc
     5761 cccgccacag tgtacctgtc taccggcaac cacgcctatt gcctgcgcta cttcgagtgg
     5821 ctgctgtctt gccccgtgat cctgatcaag ctgagcaacc tgagcggcct gaagaacgac
     5881 tacagcaagc ggaccatggg cctgatcgtg tcttgcgtgg gaatgatcgt gttcggcatg
     5941 gccgcaggac tggctaccga ttggctcaag tggctgctgt atatcgtgtc ttgcatctac
     6001 ggcggctaca tgtacttcca ggccgccaag tgctacgtgg aagccaacca cagcgtgcct
     6061 aaaggccatt gccgcatggt cgtgaagctg atggcctacg cttacttcgc ctcttggggc
     6121 agctacccaa tcctctgggc agtgggacca gaaggactgc tgaagctgag cccttacgcc
     6181 aacagcatcg gccacagcat ctgcgacatc atcgccaagg agttttggac cttcctggcc
     6241 caccacctga ggatcaagat ccacgagcac atcctgatcc acggcgacat ccggaagacc
     6301 accaagatgg agatcggagg cgaggaggtg gaagtggaag agttcgtgga ggaggaggac
     6361 gaggacacag tgtgacgtcg acggtaccat ggtattgata tctgagctcc gcatcggccg
     6421 ctgtcatcag atcgccatct cgcgcccgtg cctctgactt ctaagtccaa ttactcttca
     6481 acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta ttatcaaaaa
     6541 acttctctta atttctttgt tttttagctt cttttaagtc acctctaaca atgaaattgt
     6601 gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt gctccctccc
     6661 cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt
     6721 ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca aaatacctta
     6781 tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt ctgggcctct
     6841 catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga atttttcaat
     6901 caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt cccctattgt
     6961 ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg atgagcacga
     7021 tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga gtagaagttg
     7081 ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag aatacattcc
     7141 caatatacca aacataactg tttctactag tcggccgtac gggccctttc gtctcgcgcg
     7201 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     7261 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     7321 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     7381 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg
     7441 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     7501 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     8041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     8101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     8161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     8221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     8281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     8341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     8401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     8461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     8521 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     8581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     8641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     8701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     8761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     8821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     8881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     8941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     9001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     9061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     9121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     9181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     9241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     9301 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     9361 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     9421 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     9481 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     9541 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     9601 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     9661 acgccaagct tgcatgctcc cggccgccat ggccgcg
//
