LOCUS       sequence-351553-        7632 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 7632)
  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 7632)
  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: hSyn-somBiPOLES-mCerulean
FEATURES             Location/Qualifiers
     source          1..7632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     complement(1460..1481)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1721..1740)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2068..2089
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2489..3532
                     /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    3755..4343
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3808..3828)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4226..4237)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4384..4591
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4761..4890
                     /label=AAV2 ITR
     rep_origin      complement(5127..5715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5207..5226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5886..6677,6678..6746))
                     /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     6509..6528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6747..6851)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6877..7332)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7014..7035)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7226..7245
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7404..7533
                     /label=AAV2 ITR
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaggatcc atggcatcac aggtcgtcta
      481 tggggaatgg gcatcaactc acaccgagtg ctacaatatg tccagaatcg actccacctt
      541 cgtctcactg ctgcagctgg tgtgggccgt ggtctcaggc tgccagacaa tcttcatgat
      601 tagcagggct cccaaggtcc cttgggagtc cgtgtacctg ccctttgtcg aatctatcac
      661 ttatgcactg gccagtactg gcaacggaac cctgcagatg agggacggca gattctttcc
      721 ttggagccga atggcctcct ggctgtgcac ctgtccaatc atgctgggac agatttctaa
      781 catggctctg gtgaagtaca aaagtatccc actgaatccc attgcacagg ccgcttccat
      841 cattagagtg gtcatgggaa tcaccgctac aatttctccc gcagagtata tgaagtggct
      901 gttctttttc tttggggcca cctgcctggt gttcgagtac agtgtggtct tcacaatctt
      961 tcaagtgggg ctgtatggct ttgaatctgt cgggacacct ctggcacaga aggtggtggt
     1021 gaggatcaag atgctgcgcc tgatcttctt tattgcttgg actatgttcc caatcgtgtg
     1081 gctgatttca cccaccggag tctgcgtgat ccacgagaac gtgagcgcca ttctgtacct
     1141 gctggctgac ggcctgtgca aaaatacata tggagtgatc ctgtggagca ctgcctgggg
     1201 agtgctggaa gggaagtggg accccgcatg tctgccaggg caggagaaac ccgaagccga
     1261 cgatcctttt ggcctgaatc acgagaagaa tgcccctcct aacgacgaag tcaatatcag
     1321 aatgtttgga aggggacgta cggcggccaa gagcaggatc accagcgagg gcgagtacat
     1381 ccccctggac cagatcgaca tcaacgtggc ggtacccgtg agcaagggcg aggagctgtt
     1441 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1501 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1561 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga gctggggcgt
     1621 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1681 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1741 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1801 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacg ccatccacgg
     1861 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcggcctcaa
     1921 ctgcaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1981 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     2041 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2101 gatcactctc ggcatggacg agctgtatac cggtgcagcc ctgcaggaga agaagtcatg
     2161 cagccagcgc atggccgaat tccggcaata ctgttggaac ccggacactg ggcagatgct
     2221 gggccgcacc ccagcccggt gggtgtggat cagcctgtac tatgcagctt tctacgtggt
     2281 catgactggg ctctttgcct tgtgcatcta tgtgctgatg cagaccattg atccctacac
     2341 ccccgactac caggaccagt taaagtcacc gggggtaacc ttgagaccgg atgtgtatgg
     2401 ggaaagaggg ctgcagattt cctacaacat ctctgaaaac agctctagac aggcccagat
     2461 caccggacgt ccggagggag ctgtacaaga gctgatcagc agcgccacca gatctctgtt
     2521 tgccgccgga ggcatcaacc cttggcctaa cccctaccac cacgaggaca tgggctgtgg
     2581 aggaatgaca cctacaggcg agtgcttcag caccgagtgg tggtgtgacc cttcttacgg
     2641 actgagcgac gccggatacg gatattgctt cgtggaggcc acaggcggct acctggtcgt
     2701 gggagtggag aagaagcagg cttggctgca cagcagaggc acaccaggag aaaagatcgg
     2761 cgcccaggtc tgccagtgga ttgctttcag catcgccatc gccctgctga cattctacgg
     2821 cttcagcgct tggaaggcca cttgcggttg ggaggaggtc tacgtctgtt gcgtcgaggt
     2881 gctgttcgtg accctggaga tcttcaagga gttcagcagc cccgccacag tgtacctgtc
     2941 taccggcaac cacgcctatt gcctgcgcta cttcgagtgg ctgctgtctt gccccgtgat
     3001 cctgatcaag ctgagcaacc tgagcggcct gaagaacgac tacagcaagc ggaccatggg
     3061 cctgatcgtg tcttgcgtgg gaatgatcgt gttcggcatg gccgcaggac tggctaccga
     3121 ttggctcaag tggctgctgt atatcgtgtc ttgcatctac ggcggctaca tgtacttcca
     3181 ggccgccaag tgctacgtgg aagccaacca cagcgtgcct aaaggccatt gccgcatggt
     3241 cgtgaagctg atggcctacg cttacttcgc ctcttggggc agctacccaa tcctctgggc
     3301 agtgggacca gaaggactgc tgaagctgag cccttacgcc aacagcatcg gccacagcat
     3361 ctgcgacatc atcgccaagg agttttggac cttcctggcc caccacctga ggatcaagat
     3421 ccacgagcac atcctgatcc acggcgacat ccggaagacc accaagatgg agatcggagg
     3481 cgaggaggtg gaagtggaag agttcgtgga ggaggaggac gaggacacag tgctcgagca
     3541 gagccagccc atcctgaaca ccaaggagat ggccccccag agcaagcccc ccgaggagct
     3601 ggagatgagc agcatgccca gccccgtggc ccccctgccc gcccgcaccg agggcgtgat
     3661 cgacatgcgc agcatgagca gcatcgacag cttcatcagc tgcgccaccg acttccccga
     3721 ggccacccgc ttctgatcta gaaagcttat cgataatcaa cctctggatt acaaaatttg
     3781 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3841 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3901 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3961 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4021 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4081 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4141 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     4201 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4261 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4321 ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc tcataaagct
     4381 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4441 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4501 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4561 attgggaaga caatagcagg catgctgggg atgcggtccg gactgtacaa ttgcattcat
     4621 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     4681 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     4741 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     4801 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4861 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     4921 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4981 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5041 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5101 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5161 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5221 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5281 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5341 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5401 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5461 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5521 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5581 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5641 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5701 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5761 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5821 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5881 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5941 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6001 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6061 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6121 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6181 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6241 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6301 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6361 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6421 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6481 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6541 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6601 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6661 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6721 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6781 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6841 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     6901 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     6961 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     7021 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     7081 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     7141 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     7201 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     7261 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     7321 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     7381 gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     7441 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     7501 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
     7561 acttatctac gtagccatgc tctaggaaga tcgtaggtta attaatctag actgcagagg
     7621 gccctgcgta tg
//
