LOCUS       sequence-466500-        9158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the optogenetic tool BiPOLES in BWMs of C. elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9158)
  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 9158)
  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: pAB27 (pmyo-3::GtACR2::mCerulean::beta-HK::Chrimson)
FEATURES             Location/Qualifiers
     source          1..9158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3487..3508)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3748..3767)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4095..4116
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4516..5562
                     /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(6598..6620)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6720..6739
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6817..6835)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6903..7007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7008..7076,7077..7868)
                     /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(7226..7245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8039..8627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8528..8547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8781..8798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8915..8936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8951..8956,8957..8974,8975..8981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8989..9005
                     /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     8994..9016
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9013..9029
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9013..9029
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaggatcccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac
     2461 atagaacatt ttcaggagga cccttggatg gcatcacagg tcgtctatgg ggaatgggca
     2521 tcaactcaca ccgagtgcta caatatgtcc agaatcgact ccaccttcgt ctcactgctg
     2581 cagctggtgt gggccgtggt ctcaggctgc cagacaatct tcatgattag cagggctccc
     2641 aaggtccctt gggagtccgt gtacctgccc tttgtcgaat ctatcactta tgcactggcc
     2701 agtactggca acggaaccct gcagatgagg gacggcagat tctttccttg gagccgaatg
     2761 gcctcctggc tgtgcacctg tccaatcatg ctgggacaga tttctaacat ggctctggtg
     2821 aagtacaaaa gtatcccact gaatcccatt gcacaggccg cttccatcat tagagtggtc
     2881 atgggaatca ccgctacaat ttctcccgca gagtatatga agtggctgtt ctttttcttt
     2941 ggggccacct gcctggtgtt cgagtacagt gtggtcttca caatctttca agtggggctg
     3001 tatggctttg aatctgtcgg gacacctctg gcacagaagg tggtggtgag gatcaagatg
     3061 ctgcgcctga tcttctttat tgcttggact atgttcccaa tcgtgtggct gatttcaccc
     3121 accggagtct gcgtgatcca cgagaacgtg agcgccattc tgtacctgct ggctgacggc
     3181 ctgtgcaaaa atacatatgg agtgatcctg tggagcactg cctggggagt gctggaaggg
     3241 aagtgggacc ccgcatgtct gccagggcag gagaaacccg aagccgacga tccttttggc
     3301 ctgaatcacg agaagaatgc ccctcctaac gacgaagtca atatcagaat gtttggaagg
     3361 ggacgtacgg cggccaagag caggatcacc agcgagggcg agtacatccc cctggaccag
     3421 atcgacatca acgtggcggt acccgtgagc aagggcgagg agctgttcac cggggtggtg
     3481 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3541 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3601 ctgcccgtgc cctggcccac cctcgtgacc accctgagct ggggcgtgca gtgcttcgcc
     3661 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3721 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3781 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3841 gacggcaaca tcctggggca caagctggag tacaacgcca tccacggcaa cgtctatatc
     3901 accgccgaca agcagaagaa cggcatcaag gccaacttcg gcctcaactg caacatcgag
     3961 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4021 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     4081 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4141 atggacgagc tgtataccgg tgcagccctg caggagaaga agtcatgcag ccagcgcatg
     4201 gccgaattcc ggcaatactg ttggaacccg gacactgggc agatgctggg ccgcacccca
     4261 gcccggtggg tgtggatcag cctgtactat gcagctttct acgtggtcat gactgggctc
     4321 tttgccttgt gcatctatgt gctgatgcag accattgatc cctacacccc cgactaccag
     4381 gaccagttaa agtcaccggg ggtaaccttg agaccggatg tgtatgggga aagagggctg
     4441 cagatttcct acaacatctc tgaaaacagc tctagacagg cccagatcac cggacgtccg
     4501 gagggagctg tacaagagct gatcagcagc gccaccagat ctctgtttgc cgccggaggc
     4561 atcaaccctt ggcctaaccc ctaccaccac gaggacatgg gctgtggagg aatgacacct
     4621 acaggcgagt gcttcagcac cgagtggtgg tgtgaccctt cttacggact gagcgacgcc
     4681 ggatacggat attgcttcgt ggaggccaca ggcggctacc tggtcgtggg agtggagaag
     4741 aagcaggctt ggctgcacag cagaggcaca ccaggagaaa agatcggcgc ccaggtctgc
     4801 cagtggattg ctttcagcat cgccatcgcc ctgctgacat tctacggctt cagcgcttgg
     4861 aaggccactt gcggttggga ggaggtctac gtctgttgcg tcgaggtgct gttcgtgacc
     4921 ctggagatct tcaaggagtt cagcagcccc gccacagtgt acctgtctac cggcaaccac
     4981 gcctattgcc tgcgctactt cgagtggctg ctgtcttgcc ccgtgatcct gatcaagctg
     5041 agcaacctga gcggcctgaa gaacgactac agcaagcgga ccatgggcct gatcgtgtct
     5101 tgcgtgggaa tgatcgtgtt cggcatggcc gcaggactgg ctaccgattg gctcaagtgg
     5161 ctgctgtata tcgtgtcttg catctacggc ggctacatgt acttccaggc cgccaagtgc
     5221 tacgtggaag ccaaccacag cgtgcctaaa ggccattgcc gcatggtcgt gaagctgatg
     5281 gcctacgctt acttcgcctc ttggggcagc tacccaatcc tctgggcagt gggaccagaa
     5341 ggactgctga agctgagccc ttacgccaac agcatcggcc acagcatctg cgacatcatc
     5401 gccaaggagt tttggacctt cctggcccac cacctgagga tcaagatcca cgagcacatc
     5461 ctgatccacg gcgacatccg gaagaccacc aagatggaga tcggaggcga ggaggtggaa
     5521 gtggaagagt tcgtggagga ggaggacgag gacacagtgt gacgtcgacg gtaccggtca
     5581 gatatcactc gaggagctcc cgagatcttc agcgctcagc attcgtagaa ttgtaagttt
     5641 aaactttctc atactaacta acgcgctcta tttaaatttt cagaattcca actgagcgcc
     5701 ggtcgctacc attaccaact tgtctggtgt caaaaataat aggggccgct gtcatcagag
     5761 taagtttaaa ctgagttcta ctaactaacg agtaatattt aaattttcag catctcgcgc
     5821 ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct
     5881 gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt
     5941 agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc
     6001 gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa
     6061 aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga
     6121 aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga
     6181 ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt
     6241 gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt
     6301 ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt
     6361 tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt
     6421 gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct
     6481 atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat aactgtttcc
     6541 tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6601 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6661 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6721 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6781 cgtaaggaga aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat
     6841 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6901 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6961 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     7021 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7081 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7141 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7201 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7261 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7321 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7381 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7441 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7501 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7561 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7621 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7681 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7741 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7801 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7861 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7921 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     8041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8161 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     8221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8701 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8761 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8821 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8881 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     8941 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     9001 aacaatttca cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat
     9061 gatcttacta actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac
     9121 tcacaacgat ggatacgcta acaacttgga aatgaaat
//
