LOCUS       sequence-308755-        8669 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 8669)
  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 8669)
  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: Ef1a-DIO-BiPOLES-mCerulean
FEATURES             Location/Qualifiers
     source          1..8669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1569..2615)
                     /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(3015..3036)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3364..3383
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3623..3644
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    4654..4687
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4738..4771
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4796..5384
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4849..4869)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5267..5278)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5416..5892
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5615..5634)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5932..6072
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5932..6061
                     /label=AAV2 ITR
     rep_origin      6147..6602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6234..6253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6444..6465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6619..6638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6738..6760
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6798..6816)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6884..6988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6989..7057,7058..7849)
                     /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(7207..7226)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8020..8608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8509..8528
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgagctttc acactgtgtc ctcgtcctcc tcctccacga actcttccac ttccacctcc
     1621 tcgcctccga tctccatctt ggtggtcttc cggatgtcgc cgtggatcag gatgtgctcg
     1681 tggatcttga tcctcaggtg gtgggccagg aaggtccaaa actccttggc gatgatgtcg
     1741 cagatgctgt ggccgatgct gttggcgtaa gggctcagct tcagcagtcc ttctggtccc
     1801 actgcccaga ggattgggta gctgccccaa gaggcgaagt aagcgtaggc catcagcttc
     1861 acgaccatgc ggcaatggcc tttaggcacg ctgtggttgg cttccacgta gcacttggcg
     1921 gcctggaagt acatgtagcc gccgtagatg caagacacga tatacagcag ccacttgagc
     1981 caatcggtag ccagtcctgc ggccatgccg aacacgatca ttcccacgca agacacgatc
     2041 aggcccatgg tccgcttgct gtagtcgttc ttcaggccgc tcaggttgct cagcttgatc
     2101 aggatcacgg ggcaagacag cagccactcg aagtagcgca ggcaataggc gtggttgccg
     2161 gtagacaggt acactgtggc ggggctgctg aactccttga agatctccag ggtcacgaac
     2221 agcacctcga cgcaacagac gtagacctcc tcccaaccgc aagtggcctt ccaagcgctg
     2281 aagccgtaga atgtcagcag ggcgatggcg atgctgaaag caatccactg gcagacctgg
     2341 gcgccgatct tttctcctgg tgtgcctctg ctgtgcagcc aagcctgctt cttctccact
     2401 cccacgacca ggtagccgcc tgtggcctcc acgaagcaat atccgtatcc ggcgtcgctc
     2461 agtccgtaag aagggtcaca ccaccactcg gtgctgaagc actcgcctgt aggtgtcatt
     2521 cctccacagc ccatgtcctc gtggtggtag gggttaggcc aagggttgat gcctccggcg
     2581 gcaaacagag atctggtggc gctgctgatc agctcttgta cagctccctc cggacgtccg
     2641 gtgatctggg cctgtctaga gctgttttca gagatgttgt aggaaatctg cagccctctt
     2701 tccccataca catccggtct caaggttacc cccggtgact ttaactggtc ctggtagtcg
     2761 ggggtgtagg gatcaatggt ctgcatcagc acatagatgc acaaggcaaa gagcccagtc
     2821 atgaccacgt agaaagctgc atagtacagg ctgatccaca cccaccgggc tggggtgcgg
     2881 cccagcatct gcccagtgtc cgggttccaa cagtattgcc ggaattcggc catgcgctgg
     2941 ctgcatgact tcttctcctg cagggctgca ccggtataca gctcgtccat gccgagagtg
     3001 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     3061 ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     3121 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     3181 cagttgaggc cgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
     3241 ttgccgtgga tggcgttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     3301 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     3361 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     3421 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     3481 tgcacgcccc agctcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     3541 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     3601 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     3661 gtgaacagct cctcgccctt gctcacgggt accgccacgt tgatgtcgat ctggtccagg
     3721 gggatgtact cgccctcgct ggtgatcctg ctcttggccg ccgtacgtcc ccttccaaac
     3781 attctgatat tgacttcgtc gttaggaggg gcattcttct cgtgattcag gccaaaagga
     3841 tcgtcggctt cgggtttctc ctgccctggc agacatgcgg ggtcccactt cccttccagc
     3901 actccccagg cagtgctcca caggatcact ccatatgtat ttttgcacag gccgtcagcc
     3961 agcaggtaca gaatggcgct cacgttctcg tggatcacgc agactccggt gggtgaaatc
     4021 agccacacga ttgggaacat agtccaagca ataaagaaga tcaggcgcag catcttgatc
     4081 ctcaccacca ccttctgtgc cagaggtgtc ccgacagatt caaagccata cagccccact
     4141 tgaaagattg tgaagaccac actgtactcg aacaccaggc aggtggcccc aaagaaaaag
     4201 aacagccact tcatatactc tgcgggagaa attgtagcgg tgattcccat gaccactcta
     4261 atgatggaag cggcctgtgc aatgggattc agtgggatac ttttgtactt caccagagcc
     4321 atgttagaaa tctgtcccag catgattgga caggtgcaca gccaggaggc cattcggctc
     4381 caaggaaaga atctgccgtc cctcatctgc agggttccgt tgccagtact ggccagtgca
     4441 taagtgatag attcgacaaa gggcaggtac acggactccc aagggacctt gggagccctg
     4501 ctaatcatga agattgtctg gcagcctgag accacggccc acaccagctg cagcagtgag
     4561 acgaaggtgg agtcgattct ggacatattg tagcactcgg tgtgagttga tgcccattcc
     4621 ccatagacga cctgtgatgc catggatcct agcataactt cgtataaagt atcctatacg
     4681 aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata
     4741 acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca
     4801 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4861 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4921 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4981 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5041 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5101 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5161 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
     5221 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5281 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     5341 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
     5401 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     5461 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     5521 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     5581 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     5641 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     5701 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     5761 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     5821 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     5881 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     5941 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     6001 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     6061 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6121 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     6181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     6241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6301 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6361 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     6481 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     6541 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     6601 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     6661 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6721 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6781 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6841 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6901 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6961 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7021 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7081 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7141 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7201 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7261 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7321 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7381 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7441 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7501 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7561 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7621 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7681 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7741 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7801 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7861 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7921 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7981 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     8041 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     8101 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     8161 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     8221 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     8281 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8341 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8401 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8461 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8521 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8581 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8641 ggccttttgc tggccttttg ctcacatgt
//
