LOCUS       sequence-308736-        7811 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 7811)
  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 7811)
  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-BiPOLES-mCerulean
FEATURES             Location/Qualifiers
     source          1..7811
                     /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"
     primer_bind     complement(2028..2049)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2289..2308)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2636..2657
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3057..4103
                     /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    4157..4745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4210..4230)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4628..4639)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4914..4995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4951..4970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5005..5024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5074..5214
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5074..5203
                     /label=AAV2 ITR
     rep_origin      5289..5744
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5376..5395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5586..5607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5761..5780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5880..5902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5940..5958)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6026..6130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6131..6199,6200..6991)
                     /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(6349..6368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7162..7750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7651..7670
                     /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 gtggatccat ggcatcacag gtcgtctatg gggaatgggc atcaactcac accgagtgct
     1081 acaatatgtc cagaatcgac tccaccttcg tctcactgct gcagctggtg tgggccgtgg
     1141 tctcaggctg ccagacaatc ttcatgatta gcagggctcc caaggtccct tgggagtccg
     1201 tgtacctgcc ctttgtcgaa tctatcactt atgcactggc cagtactggc aacggaaccc
     1261 tgcagatgag ggacggcaga ttctttcctt ggagccgaat ggcctcctgg ctgtgcacct
     1321 gtccaatcat gctgggacag atttctaaca tggctctggt gaagtacaaa agtatcccac
     1381 tgaatcccat tgcacaggcc gcttccatca ttagagtggt catgggaatc accgctacaa
     1441 tttctcccgc agagtatatg aagtggctgt tctttttctt tggggccacc tgcctggtgt
     1501 tcgagtacag tgtggtcttc acaatctttc aagtggggct gtatggcttt gaatctgtcg
     1561 ggacacctct ggcacagaag gtggtggtga ggatcaagat gctgcgcctg atcttcttta
     1621 ttgcttggac tatgttccca atcgtgtggc tgatttcacc caccggagtc tgcgtgatcc
     1681 acgagaacgt gagcgccatt ctgtacctgc tggctgacgg cctgtgcaaa aatacatatg
     1741 gagtgatcct gtggagcact gcctggggag tgctggaagg gaagtgggac cccgcatgtc
     1801 tgccagggca ggagaaaccc gaagccgacg atccttttgg cctgaatcac gagaagaatg
     1861 cccctcctaa cgacgaagtc aatatcagaa tgtttggaag gggacgtacg gcggccaaga
     1921 gcaggatcac cagcgagggc gagtacatcc ccctggacca gatcgacatc aacgtggcgg
     1981 tacccgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2041 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2101 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2161 ccctcgtgac caccctgagc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
     2221 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2281 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2341 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2401 acaagctgga gtacaacgcc atccacggca acgtctatat caccgccgac aagcagaaga
     2461 acggcatcaa ggccaacttc ggcctcaact gcaacatcga ggacggcagc gtgcagctcg
     2521 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2581 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     2641 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtataccg
     2701 gtgcagccct gcaggagaag aagtcatgca gccagcgcat ggccgaattc cggcaatact
     2761 gttggaaccc ggacactggg cagatgctgg gccgcacccc agcccggtgg gtgtggatca
     2821 gcctgtacta tgcagctttc tacgtggtca tgactgggct ctttgccttg tgcatctatg
     2881 tgctgatgca gaccattgat ccctacaccc ccgactacca ggaccagtta aagtcaccgg
     2941 gggtaacctt gagaccggat gtgtatgggg aaagagggct gcagatttcc tacaacatct
     3001 ctgaaaacag ctctagacag gcccagatca ccggacgtcc ggagggagct gtacaagagc
     3061 tgatcagcag cgccaccaga tctctgtttg ccgccggagg catcaaccct tggcctaacc
     3121 cctaccacca cgaggacatg ggctgtggag gaatgacacc tacaggcgag tgcttcagca
     3181 ccgagtggtg gtgtgaccct tcttacggac tgagcgacgc cggatacgga tattgcttcg
     3241 tggaggccac aggcggctac ctggtcgtgg gagtggagaa gaagcaggct tggctgcaca
     3301 gcagaggcac accaggagaa aagatcggcg cccaggtctg ccagtggatt gctttcagca
     3361 tcgccatcgc cctgctgaca ttctacggct tcagcgcttg gaaggccact tgcggttggg
     3421 aggaggtcta cgtctgttgc gtcgaggtgc tgttcgtgac cctggagatc ttcaaggagt
     3481 tcagcagccc cgccacagtg tacctgtcta ccggcaacca cgcctattgc ctgcgctact
     3541 tcgagtggct gctgtcttgc cccgtgatcc tgatcaagct gagcaacctg agcggcctga
     3601 agaacgacta cagcaagcgg accatgggcc tgatcgtgtc ttgcgtggga atgatcgtgt
     3661 tcggcatggc cgcaggactg gctaccgatt ggctcaagtg gctgctgtat atcgtgtctt
     3721 gcatctacgg cggctacatg tacttccagg ccgccaagtg ctacgtggaa gccaaccaca
     3781 gcgtgcctaa aggccattgc cgcatggtcg tgaagctgat ggcctacgct tacttcgcct
     3841 cttggggcag ctacccaatc ctctgggcag tgggaccaga aggactgctg aagctgagcc
     3901 cttacgccaa cagcatcggc cacagcatct gcgacatcat cgccaaggag ttttggacct
     3961 tcctggccca ccacctgagg atcaagatcc acgagcacat cctgatccac ggcgacatcc
     4021 ggaagaccac caagatggag atcggaggcg aggaggtgga agtggaagag ttcgtggagg
     4081 aggaggacga ggacacagtg tgaaagctta agtttcgcgc cacccctgca gggaattcga
     4141 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4201 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4261 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4321 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4381 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4441 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4501 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4561 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     4621 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4681 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4741 cccgcatcga taccgagcgc tgctcgagag atctgtgata gcggccatca agctggccgc
     4801 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4861 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcagcttatc
     5041 gataccgcat gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     5101 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     5161 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     5221 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     5281 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5341 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     5401 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5461 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     5521 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5581 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     5641 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     5701 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     5761 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5821 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5881 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5941 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6001 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7801 tgctcacatg t
//
