LOCUS       sequence-308722-        7425 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 7425)
  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 7425)
  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-BiPOLES-mCerulean
FEATURES             Location/Qualifiers
     source          1..7425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2688..3135
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     complement(4147..4168)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4408..4427)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4755..4776
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5176..6222
                     /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    6235..6823
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6288..6308)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6706..6717)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6864..7071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7241..7370
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa tctagactgc agagggccct gcgtatgagt gcaagtgggt
     2701 tttaggacca ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg
     2761 gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga
     2821 aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg
     2881 cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc
     2941 gccggtcccc cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc
     3001 ccagccggac cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc
     3061 tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg
     3121 tgcctgagag cgcagtcgag aggatccatg gcatcacagg tcgtctatgg ggaatgggca
     3181 tcaactcaca ccgagtgcta caatatgtcc agaatcgact ccaccttcgt ctcactgctg
     3241 cagctggtgt gggccgtggt ctcaggctgc cagacaatct tcatgattag cagggctccc
     3301 aaggtccctt gggagtccgt gtacctgccc tttgtcgaat ctatcactta tgcactggcc
     3361 agtactggca acggaaccct gcagatgagg gacggcagat tctttccttg gagccgaatg
     3421 gcctcctggc tgtgcacctg tccaatcatg ctgggacaga tttctaacat ggctctggtg
     3481 aagtacaaaa gtatcccact gaatcccatt gcacaggccg cttccatcat tagagtggtc
     3541 atgggaatca ccgctacaat ttctcccgca gagtatatga agtggctgtt ctttttcttt
     3601 ggggccacct gcctggtgtt cgagtacagt gtggtcttca caatctttca agtggggctg
     3661 tatggctttg aatctgtcgg gacacctctg gcacagaagg tggtggtgag gatcaagatg
     3721 ctgcgcctga tcttctttat tgcttggact atgttcccaa tcgtgtggct gatttcaccc
     3781 accggagtct gcgtgatcca cgagaacgtg agcgccattc tgtacctgct ggctgacggc
     3841 ctgtgcaaaa atacatatgg agtgatcctg tggagcactg cctggggagt gctggaaggg
     3901 aagtgggacc ccgcatgtct gccagggcag gagaaacccg aagccgacga tccttttggc
     3961 ctgaatcacg agaagaatgc ccctcctaac gacgaagtca atatcagaat gtttggaagg
     4021 ggacgtacgg cggccaagag caggatcacc agcgagggcg agtacatccc cctggaccag
     4081 atcgacatca acgtggcggt acccgtgagc aagggcgagg agctgttcac cggggtggtg
     4141 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4201 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4261 ctgcccgtgc cctggcccac cctcgtgacc accctgagct ggggcgtgca gtgcttcgcc
     4321 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4381 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     4441 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4501 gacggcaaca tcctggggca caagctggag tacaacgcca tccacggcaa cgtctatatc
     4561 accgccgaca agcagaagaa cggcatcaag gccaacttcg gcctcaactg caacatcgag
     4621 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4681 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     4741 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4801 atggacgagc tgtataccgg tgcagccctg caggagaaga agtcatgcag ccagcgcatg
     4861 gccgaattcc ggcaatactg ttggaacccg gacactgggc agatgctggg ccgcacccca
     4921 gcccggtggg tgtggatcag cctgtactat gcagctttct acgtggtcat gactgggctc
     4981 tttgccttgt gcatctatgt gctgatgcag accattgatc cctacacccc cgactaccag
     5041 gaccagttaa agtcaccggg ggtaaccttg agaccggatg tgtatgggga aagagggctg
     5101 cagatttcct acaacatctc tgaaaacagc tctagacagg cccagatcac cggacgtccg
     5161 gagggagctg tacaagagct gatcagcagc gccaccagat ctctgtttgc cgccggaggc
     5221 atcaaccctt ggcctaaccc ctaccaccac gaggacatgg gctgtggagg aatgacacct
     5281 acaggcgagt gcttcagcac cgagtggtgg tgtgaccctt cttacggact gagcgacgcc
     5341 ggatacggat attgcttcgt ggaggccaca ggcggctacc tggtcgtggg agtggagaag
     5401 aagcaggctt ggctgcacag cagaggcaca ccaggagaaa agatcggcgc ccaggtctgc
     5461 cagtggattg ctttcagcat cgccatcgcc ctgctgacat tctacggctt cagcgcttgg
     5521 aaggccactt gcggttggga ggaggtctac gtctgttgcg tcgaggtgct gttcgtgacc
     5581 ctggagatct tcaaggagtt cagcagcccc gccacagtgt acctgtctac cggcaaccac
     5641 gcctattgcc tgcgctactt cgagtggctg ctgtcttgcc ccgtgatcct gatcaagctg
     5701 agcaacctga gcggcctgaa gaacgactac agcaagcgga ccatgggcct gatcgtgtct
     5761 tgcgtgggaa tgatcgtgtt cggcatggcc gcaggactgg ctaccgattg gctcaagtgg
     5821 ctgctgtata tcgtgtcttg catctacggc ggctacatgt acttccaggc cgccaagtgc
     5881 tacgtggaag ccaaccacag cgtgcctaaa ggccattgcc gcatggtcgt gaagctgatg
     5941 gcctacgctt acttcgcctc ttggggcagc tacccaatcc tctgggcagt gggaccagaa
     6001 ggactgctga agctgagccc ttacgccaac agcatcggcc acagcatctg cgacatcatc
     6061 gccaaggagt tttggacctt cctggcccac cacctgagga tcaagatcca cgagcacatc
     6121 ctgatccacg gcgacatccg gaagaccacc aagatggaga tcggaggcga ggaggtggaa
     6181 gtggaagagt tcgtggagga ggaggacgag gacacagtgt gaaagcttat cgataatcaa
     6241 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6301 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6361 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6421 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6481 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6541 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6601 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     6661 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6721 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6781 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc
     6841 gggcggccgc tcataaagct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     6901 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     6961 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     7021 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtccg
     7081 gactgtacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     7141 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc
     7201 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     7261 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     7321 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat
     7381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgct
//
