LOCUS       sequence-308732-        8020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic tool for blue-light inhibition and red-light excitation 
            of neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8020)
  AUTHORS   Vierock J, Rodriguez-Rozada S, Dieter A, Pieper F, Sims R, Tenedini 
            F, Bergs ACF, Bendifallah I, Zhou F, Zeitzschel N, Ahlbeck J, 
            Augustin S, Sauter K, Papagiakoumou E, Gottschalk A, Soba P, 
            Emiliani V, Engel AK, Hegemann P, Wiegert JS
  TITLE     BiPOLES is an optogenetic tool developed for bidirectional 
            dual-color control of neurons.
  JOURNAL   Nat Commun. 2021 Jul 26;12(1):4527. doi: 10.1038/s41467-021-24759-5.
  PUBMED    34312384
REFERENCE   2  (bases 1 to 8020)
  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: "2021-07-26"; volume: "12(1):4527. doi"; pages: " 
            10.1038/s41467-021-24759-5"
COMMENT     Sequence Label: mDlx-somBiPOLES-mCerulean
FEATURES             Location/Qualifiers
     source          1..8020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          849..981
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     964..983
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1024..1033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2037..2058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2298..2317)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2645..2666
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3066..4109
                     /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"
     misc_feature    4366..4954
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4419..4439)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4837..4848)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5123..5204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5160..5179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5214..5233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5283..5423
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5283..5412
                     /label=AAV2 ITR
     rep_origin      5498..5953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5585..5604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5795..5816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5970..5989)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6089..6111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6149..6167)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6235..6339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6340..6408,6409..7200)
                     /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(6558..6577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7371..7959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7860..7879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aagaattcta tacactcaca gtggtttggc
      181 atatatttgg tgaaattttt taaggaaaaa ttagtgttgg tttcgatata tggtagcttt
      241 ttctctaaca taatttgaat aattcagcaa agccctacta ccagctgtac ttctgcagcc
      301 tcttccattc tttccagcat tataattttg gttaattttc aattttaggt cctacgtctc
      361 tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct
      421 gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc
      481 tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg
      541 catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt
      601 ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc
      661 tacgaaccca aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg
      721 cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg
      781 agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca
      841 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      901 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      961 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taagaaacta
     1021 gtagccacca tggatccatg gcatcacagg tcgtctatgg ggaatgggca tcaactcaca
     1081 ccgagtgcta caatatgtcc agaatcgact ccaccttcgt ctcactgctg cagctggtgt
     1141 gggccgtggt ctcaggctgc cagacaatct tcatgattag cagggctccc aaggtccctt
     1201 gggagtccgt gtacctgccc tttgtcgaat ctatcactta tgcactggcc agtactggca
     1261 acggaaccct gcagatgagg gacggcagat tctttccttg gagccgaatg gcctcctggc
     1321 tgtgcacctg tccaatcatg ctgggacaga tttctaacat ggctctggtg aagtacaaaa
     1381 gtatcccact gaatcccatt gcacaggccg cttccatcat tagagtggtc atgggaatca
     1441 ccgctacaat ttctcccgca gagtatatga agtggctgtt ctttttcttt ggggccacct
     1501 gcctggtgtt cgagtacagt gtggtcttca caatctttca agtggggctg tatggctttg
     1561 aatctgtcgg gacacctctg gcacagaagg tggtggtgag gatcaagatg ctgcgcctga
     1621 tcttctttat tgcttggact atgttcccaa tcgtgtggct gatttcaccc accggagtct
     1681 gcgtgatcca cgagaacgtg agcgccattc tgtacctgct ggctgacggc ctgtgcaaaa
     1741 atacatatgg agtgatcctg tggagcactg cctggggagt gctggaaggg aagtgggacc
     1801 ccgcatgtct gccagggcag gagaaacccg aagccgacga tccttttggc ctgaatcacg
     1861 agaagaatgc ccctcctaac gacgaagtca atatcagaat gtttggaagg ggacgtacgg
     1921 cggccaagag caggatcacc agcgagggcg agtacatccc cctggaccag atcgacatca
     1981 acgtggcggt acccgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2041 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2101 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2161 cctggcccac cctcgtgacc accctgagct ggggcgtgca gtgcttcgcc cgctaccccg
     2221 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2281 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2341 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2401 tcctggggca caagctggag tacaacgcca tccacggcaa cgtctatatc accgccgaca
     2461 agcagaagaa cggcatcaag gccaacttcg gcctcaactg caacatcgag gacggcagcg
     2521 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2581 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     2641 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2701 tgtataccgg tgcagccctg caggagaaga agtcatgcag ccagcgcatg gccgaattcc
     2761 ggcaatactg ttggaacccg gacactgggc agatgctggg ccgcacccca gcccggtggg
     2821 tgtggatcag cctgtactat gcagctttct acgtggtcat gactgggctc tttgccttgt
     2881 gcatctatgt gctgatgcag accattgatc cctacacccc cgactaccag gaccagttaa
     2941 agtcaccggg ggtaaccttg agaccggatg tgtatgggga aagagggctg cagatttcct
     3001 acaacatctc tgaaaacagc tctagacagg cccagatcac cggacgtccg gagggagctg
     3061 tacaagagct gatcagcagc gccaccagat ctctgtttgc cgccggaggc atcaaccctt
     3121 ggcctaaccc ctaccaccac gaggacatgg gctgtggagg aatgacacct acaggcgagt
     3181 gcttcagcac cgagtggtgg tgtgaccctt cttacggact gagcgacgcc ggatacggat
     3241 attgcttcgt ggaggccaca ggcggctacc tggtcgtggg agtggagaag aagcaggctt
     3301 ggctgcacag cagaggcaca ccaggagaaa agatcggcgc ccaggtctgc cagtggattg
     3361 ctttcagcat cgccatcgcc ctgctgacat tctacggctt cagcgcttgg aaggccactt
     3421 gcggttggga ggaggtctac gtctgttgcg tcgaggtgct gttcgtgacc ctggagatct
     3481 tcaaggagtt cagcagcccc gccacagtgt acctgtctac cggcaaccac gcctattgcc
     3541 tgcgctactt cgagtggctg ctgtcttgcc ccgtgatcct gatcaagctg agcaacctga
     3601 gcggcctgaa gaacgactac agcaagcgga ccatgggcct gatcgtgtct tgcgtgggaa
     3661 tgatcgtgtt cggcatggcc gcaggactgg ctaccgattg gctcaagtgg ctgctgtata
     3721 tcgtgtcttg catctacggc ggctacatgt acttccaggc cgccaagtgc tacgtggaag
     3781 ccaaccacag cgtgcctaaa ggccattgcc gcatggtcgt gaagctgatg gcctacgctt
     3841 acttcgcctc ttggggcagc tacccaatcc tctgggcagt gggaccagaa ggactgctga
     3901 agctgagccc ttacgccaac agcatcggcc acagcatctg cgacatcatc gccaaggagt
     3961 tttggacctt cctggcccac cacctgagga tcaagatcca cgagcacatc ctgatccacg
     4021 gcgacatccg gaagaccacc aagatggaga tcggaggcga ggaggtggaa gtggaagagt
     4081 tcgtggagga ggaggacgag gacacagtgc tcgagcagag ccagcccatc ctgaacacca
     4141 aggagatggc cccccagagc aagccccccg aggagctgga gatgagcagc atgcccagcc
     4201 ccgtggcccc cctgcccgcc cgcaccgagg gcgtgatcga catgcgcagc atgagcagca
     4261 tcgacagctt catcagctgc gccaccgact tccccgaggc cacccgcttc tgatctagaa
     4321 agctcgcgcc acccctgcag ggaattcgat atcaagctta tcgataatca acctctggat
     4381 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4441 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4501 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     4561 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     4621 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     4681 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4741 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     4801 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4861 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4921 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     4981 tctgtgatag cggccatcaa gctggccgcg actctagatc ataatcagcc ataccacatt
     5041 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     5101 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     5161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5221 gtccaaactc atcaatgtat cagcttatcg ataccgcatg cacgtgcgga ccgagcggcc
     5281 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     5341 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     5401 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     5461 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     5521 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     5581 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5641 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5701 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5761 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5821 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     5881 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5941 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6001 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6061 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6121 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     6181 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6241 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6301 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6361 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6421 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6481 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6541 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6601 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6661 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6721 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6781 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6841 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6901 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6961 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7021 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7081 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7141 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7201 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7261 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7321 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7381 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7441 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7501 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7561 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7621 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7681 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7741 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7801 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7861 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7921 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7981 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
