LOCUS       sequence-466450-        8013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the optogenetic tool BiPOLES in GABAergic neurons of C. 
            elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8013)
  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 8013)
  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: pAB31 (punc-47::GtACR2::mCerulean::bHK::Chrimson)
FEATURES             Location/Qualifiers
     source          1..8013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2449..2470)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2710..2729)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3057..3078
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3478..4524
                     /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(5435..5457)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5557..5576
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5654..5672)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5740..5844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5845..5913,5914..6705)
                     /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(6063..6082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6876..7464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7365..7384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7618..7635
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7752..7773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7788..7793,7794..7811,7812..7818)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7826..7842
                     /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     7831..7853
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7850..7866
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7850..7866
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agcttgttgt catcacttca aacttttcaa tgcataaatt tcttgtcaga tctgatcctc
       61 tatacttgaa gtttataccg ataaaggttt tcaacagatt ttgaagcatt tgaatcaggg
      121 gtgcctgggt aaacctatca tacgaaactg ccaatttgtc ctgtgaatcg tatctattaa
      181 aacaagactg aaaataacta aacttctacg tcaaaaagtt gacaaaacaa ctttcttgga
      241 aaaaaagctg ttgggatccc ggaacagtcg aaagtcggtg gcaagcgccg aactgctgac
      301 ggtctaaccg gggcacaaat caggggtgag cggcaaacga tttttccggc aaatcggcaa
      361 atcggcaaat tgccaatatt gaaatacccg gcaaatcggt aaatagccgg aattgaaaat
      421 ttccggcaaa ctggtaaacc gcaaattgct gatttgccga atttgccggg aagacggcaa
      481 ttgccaaaca tattcggcaa attgtggttt tgcacttttt ggaaatttca gaatttcaat
      541 cggcaaattg tgcacatcct atgaattttc ctacgtctat tttgaaaagt aagcaaattc
      601 tatgaaaata tctaaagaaa aatggaaaaa attttcaaaa aggcacagtt ttaagtgttt
      661 ccgtctaata aaaaaatccc cctaaacact tccggcaaat tgatgttcgg caaatggcaa
      721 atcggaaact tgccgaaaat tacagtttcc ggtaaatcgg caaaccggca aactgcctga
      781 attgaaaagt tccgtcaaat cggcaaaccg acaacacccc tggcacaaat gatggacata
      841 ctgaggcaat ttgccggttt tccaattgca ggaaattttc aattccggca gtgtgccgat
      901 ttgccggaaa ttttaattca ggcaaattgc cgatttcccg atttcccgat ttgccggaaa
      961 aaatcgtttg ccgcccaccc ctgggtctga accttgattg ttacaaaaca tttttagctc
     1021 tttggagaaa taaaatgaat ctcgtaaaat ttaattgacg aggacgatat tagctgtctc
     1081 tttagaccaa attcagaaaa aaagaaagaa tacttcccaa atttccggtc cctctctcgt
     1141 tttttttgcc aataaactca ctatagtcgc tggttccccc ctattcacat ttattctacc
     1201 aatccatcag tggaaccaga aaaaagaaga gcctttcggt ttggagagta gggtctaata
     1261 atcccccgtg ctcttcaaat cattgtgcca acacacagac acactttatg tgtgctcaca
     1321 cacacacgct atttgaagag cgaagacgac gacgacgcat tcagagctct tttccacgaa
     1381 atttgctcca tctttccaca atctgtcttt cctgtgagac gacagcgtca catttatttc
     1441 attacagcaa tggcatcaca ggtcgtctat ggggaatggg catcaactca caccgagtgc
     1501 tacaatatgt ccagaatcga ctccaccttc gtctcactgc tgcagctggt gtgggccgtg
     1561 gtctcaggct gccagacaat cttcatgatt agcagggctc ccaaggtccc ttgggagtcc
     1621 gtgtacctgc cctttgtcga atctatcact tatgcactgg ccagtactgg caacggaacc
     1681 ctgcagatga gggacggcag attctttcct tggagccgaa tggcctcctg gctgtgcacc
     1741 tgtccaatca tgctgggaca gatttctaac atggctctgg tgaagtacaa aagtatccca
     1801 ctgaatccca ttgcacaggc cgcttccatc attagagtgg tcatgggaat caccgctaca
     1861 atttctcccg cagagtatat gaagtggctg ttctttttct ttggggccac ctgcctggtg
     1921 ttcgagtaca gtgtggtctt cacaatcttt caagtggggc tgtatggctt tgaatctgtc
     1981 gggacacctc tggcacagaa ggtggtggtg aggatcaaga tgctgcgcct gatcttcttt
     2041 attgcttgga ctatgttccc aatcgtgtgg ctgatttcac ccaccggagt ctgcgtgatc
     2101 cacgagaacg tgagcgccat tctgtacctg ctggctgacg gcctgtgcaa aaatacatat
     2161 ggagtgatcc tgtggagcac tgcctgggga gtgctggaag ggaagtggga ccccgcatgt
     2221 ctgccagggc aggagaaacc cgaagccgac gatccttttg gcctgaatca cgagaagaat
     2281 gcccctccta acgacgaagt caatatcaga atgtttggaa ggggacgtac ggcggccaag
     2341 agcaggatca ccagcgaggg cgagtacatc cccctggacc agatcgacat caacgtggcg
     2401 gtacccgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2461 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2521 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2581 accctcgtga ccaccctgag ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg
     2641 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2701 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2761 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2821 cacaagctgg agtacaacgc catccacggc aacgtctata tcaccgccga caagcagaag
     2881 aacggcatca aggccaactt cggcctcaac tgcaacatcg aggacggcag cgtgcagctc
     2941 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3001 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     3061 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtatacc
     3121 ggtgcagccc tgcaggagaa gaagtcatgc agccagcgca tggccgaatt ccggcaatac
     3181 tgttggaacc cggacactgg gcagatgctg ggccgcaccc cagcccggtg ggtgtggatc
     3241 agcctgtact atgcagcttt ctacgtggtc atgactgggc tctttgcctt gtgcatctat
     3301 gtgctgatgc agaccattga tccctacacc cccgactacc aggaccagtt aaagtcaccg
     3361 ggggtaacct tgagaccgga tgtgtatggg gaaagagggc tgcagatttc ctacaacatc
     3421 tctgaaaaca gctctagaca ggcccagatc accggacgtc cggagggagc tgtacaagag
     3481 ctgatcagca gcgccaccag atctctgttt gccgccggag gcatcaaccc ttggcctaac
     3541 ccctaccacc acgaggacat gggctgtgga ggaatgacac ctacaggcga gtgcttcagc
     3601 accgagtggt ggtgtgaccc ttcttacgga ctgagcgacg ccggatacgg atattgcttc
     3661 gtggaggcca caggcggcta cctggtcgtg ggagtggaga agaagcaggc ttggctgcac
     3721 agcagaggca caccaggaga aaagatcggc gcccaggtct gccagtggat tgctttcagc
     3781 atcgccatcg ccctgctgac attctacggc ttcagcgctt ggaaggccac ttgcggttgg
     3841 gaggaggtct acgtctgttg cgtcgaggtg ctgttcgtga ccctggagat cttcaaggag
     3901 ttcagcagcc ccgccacagt gtacctgtct accggcaacc acgcctattg cctgcgctac
     3961 ttcgagtggc tgctgtcttg ccccgtgatc ctgatcaagc tgagcaacct gagcggcctg
     4021 aagaacgact acagcaagcg gaccatgggc ctgatcgtgt cttgcgtggg aatgatcgtg
     4081 ttcggcatgg ccgcaggact ggctaccgat tggctcaagt ggctgctgta tatcgtgtct
     4141 tgcatctacg gcggctacat gtacttccag gccgccaagt gctacgtgga agccaaccac
     4201 agcgtgccta aaggccattg ccgcatggtc gtgaagctga tggcctacgc ttacttcgcc
     4261 tcttggggca gctacccaat cctctgggca gtgggaccag aaggactgct gaagctgagc
     4321 ccttacgcca acagcatcgg ccacagcatc tgcgacatca tcgccaagga gttttggacc
     4381 ttcctggccc accacctgag gatcaagatc cacgagcaca tcctgatcca cggcgacatc
     4441 cggaagacca ccaagatgga gatcggaggc gaggaggtgg aagtggaaga gttcgtggag
     4501 gaggaggacg aggacacagt gtgaccaact gagcgccggt cgctaccatt accaacttgt
     4561 ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa gtttaaactg agttctacta
     4621 actaacgagt aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc
     4681 aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt
     4741 tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag tcacctctaa
     4801 caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt
     4861 gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac
     4921 acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca
     4981 caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac
     5041 gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg
     5101 gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt
     5161 ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat
     5221 tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt
     5281 gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac
     5341 agaatacatt cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct
     5401 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5461 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5521 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5581 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     5641 ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5701 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5761 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5821 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5881 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5941 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6001 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6061 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6121 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6181 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6241 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6301 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6361 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6421 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6481 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6541 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6601 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6661 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6721 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6781 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6841 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6961 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7021 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7081 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7141 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7201 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7261 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7321 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7381 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7441 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7501 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7561 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7621 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7681 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7741 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7801 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7861 tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc
     7921 atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca
     7981 acttggaaat gaaataagct tgcatgcctg cag
//
