LOCUS       sequence-299906-        7052 bp    DNA     circular SYN 14-APR-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 7052)
  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 7052)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: CaMKII-somBiPOLES-mCerulean
FEATURES             Location/Qualifiers
     source          1..7052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     primer_bind     complement(1538..1559)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1799..1818)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2146..2167
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2567..3610
                     /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    3827..4230
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    4237..4285
                     /note="synthetic polyadenylation signal"
     repeat_region   4315..4455
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4315..4444
                     /label=AAV2 ITR
     rep_origin      4530..4985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4827..4848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5002..5021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5121..5143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5181..5199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5267..5371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5372..5440,5441..6232)
                     /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(5590..5609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6403..6991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6892..6911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcttgt ggactaagtt tgttcgcatc
      181 cccttctcca accccctcag tacatcaccc tgggggaaca gggtccactt gctcctgggc
      241 ccacacagtc ctgcagtatt gtgtatataa ggccagggca aagaggagca ggttttaaag
      301 tgaaaggcag gcaggtgttg gggaggcagt taccggggca acgggaacag ggcgtttcgg
      361 aggtggttgc catggggacc tggatgctga cgaaggctcg cgaggctgtg agcagccaca
      421 gtgccctgct cagaagcccc aagctcgtca gtcaagccgg ttctccgttt gcactcagga
      481 gcacgggcag gcgagtggcc cctagttctg ggggcagctc tagagcggta ccggatccat
      541 ggcatcacag gtcgtctatg gggaatgggc atcaactcac accgagtgct acaatatgtc
      601 cagaatcgac tccaccttcg tctcactgct gcagctggtg tgggccgtgg tctcaggctg
      661 ccagacaatc ttcatgatta gcagggctcc caaggtccct tgggagtccg tgtacctgcc
      721 ctttgtcgaa tctatcactt atgcactggc cagtactggc aacggaaccc tgcagatgag
      781 ggacggcaga ttctttcctt ggagccgaat ggcctcctgg ctgtgcacct gtccaatcat
      841 gctgggacag atttctaaca tggctctggt gaagtacaaa agtatcccac tgaatcccat
      901 tgcacaggcc gcttccatca ttagagtggt catgggaatc accgctacaa tttctcccgc
      961 agagtatatg aagtggctgt tctttttctt tggggccacc tgcctggtgt tcgagtacag
     1021 tgtggtcttc acaatctttc aagtggggct gtatggcttt gaatctgtcg ggacacctct
     1081 ggcacagaag gtggtggtga ggatcaagat gctgcgcctg atcttcttta ttgcttggac
     1141 tatgttccca atcgtgtggc tgatttcacc caccggagtc tgcgtgatcc acgagaacgt
     1201 gagcgccatt ctgtacctgc tggctgacgg cctgtgcaaa aatacatatg gagtgatcct
     1261 gtggagcact gcctggggag tgctggaagg gaagtgggac cccgcatgtc tgccagggca
     1321 ggagaaaccc gaagccgacg atccttttgg cctgaatcac gagaagaatg cccctcctaa
     1381 cgacgaagtc aatatcagaa tgtttggaag gggacgtacg gcggccaaga gcaggatcac
     1441 cagcgagggc gagtacatcc ccctggacca gatcgacatc aacgtggcgg tacccgtgag
     1501 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1561 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1621 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1681 caccctgagc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1741 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1801 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1861 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1921 gtacaacgcc atccacggca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1981 ggccaacttc ggcctcaact gcaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2041 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2101 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2161 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtataccg gtgcagccct
     2221 gcaggagaag aagtcatgca gccagcgcat ggccgaattc cggcaatact gttggaaccc
     2281 ggacactggg cagatgctgg gccgcacccc agcccggtgg gtgtggatca gcctgtacta
     2341 tgcagctttc tacgtggtca tgactgggct ctttgccttg tgcatctatg tgctgatgca
     2401 gaccattgat ccctacaccc ccgactacca ggaccagtta aagtcaccgg gggtaacctt
     2461 gagaccggat gtgtatgggg aaagagggct gcagatttcc tacaacatct ctgaaaacag
     2521 ctctagacag gcccagatca ccggacgtcc ggagggagct gtacaagagc tgatcagcag
     2581 cgccaccaga tctctgtttg ccgccggagg catcaaccct tggcctaacc cctaccacca
     2641 cgaggacatg ggctgtggag gaatgacacc tacaggcgag tgcttcagca ccgagtggtg
     2701 gtgtgaccct tcttacggac tgagcgacgc cggatacgga tattgcttcg tggaggccac
     2761 aggcggctac ctggtcgtgg gagtggagaa gaagcaggct tggctgcaca gcagaggcac
     2821 accaggagaa aagatcggcg cccaggtctg ccagtggatt gctttcagca tcgccatcgc
     2881 cctgctgaca ttctacggct tcagcgcctg gaaggccact tgcggttggg aggaggtcta
     2941 cgtctgttgc gtcgaggtgc tgttcgtgac cctggagatc ttcaaggagt tcagcagccc
     3001 cgccacagtg tacctgtcta ccggcaacca cgcctattgc ctgcgctact tcgagtggct
     3061 gctgtcttgc cccgtgatcc tgatcaagct gagcaacctg agcggcctga agaacgacta
     3121 cagcaagcgg accatgggcc tgatcgtgtc ttgcgtggga atgatcgtgt tcggcatggc
     3181 cgcaggactg gctaccgatt ggctcaagtg gctgctgtat atcgtgtctt gcatctacgg
     3241 cggctacatg tacttccagg ccgccaagtg ctacgtggaa gccaaccaca gcgtgcctaa
     3301 aggccattgc cgcatggtcg tgaagctgat ggcctacgct tacttcgcct cttggggcag
     3361 ctacccaatc ctctgggcag tgggaccaga aggactgctg aagctgagcc cttacgccaa
     3421 cagcatcggc cacagcatct gcgacatcat cgccaaggag ttttggacct tcctggccca
     3481 ccacctgagg atcaagatcc acgagcacat cctgatccac ggcgacatcc ggaagaccac
     3541 caagatggag atcggaggcg aggaggtgga agtggaagag ttcgtggagg aggaggacga
     3601 ggacacagtg ctcgagcaga gccagcccat cctgaacacc aaggagatgg ccccccagag
     3661 caagcccccc gaggagctgg agatgagcag catgcccagc cccgtggccc ccctgcccgc
     3721 ccgcaccgag ggcgtgatcg acatgcgcag catgagcagc atcgacagct tcatcagctg
     3781 cgccaccgac ttccccgagg ccacccgctt ctgatctaga aagcttaatc aacctctgga
     3841 ttacaaaatt tgtgaaagat tgactgatat tcttaactat gttgctcctt ttacgctgtg
     3901 tggatatgct gctttaatgc ctctgtatca tgctattgct tcccgtatgg ctttcgtttt
     3961 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtccg
     4021 tcaacgtggc gtggtgtgct ctgtgtttgc tgacgcaacc cccactggct ggggcattgc
     4081 caccacctgt caactccttt ctgggacttt cgctttcccc ctcccgatcg ccacggcaga
     4141 actcatcgcc gcctgccttg cccgctgctg gacaggggct aggttgctgg gcactgataa
     4201 ttccgtggtg ttgtcgggga agctgacgtc agcgctaata aaatcgctat ccatcgaaga
     4261 tggatgtgtg ttggtttttt gtgtgggtaa cccacgtgcg gaccgagcgg ccgcaggaac
     4321 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4381 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4441 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4501 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4561 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4621 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4681 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4741 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4801 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4861 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4921 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4981 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5041 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5101 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5161 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5221 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5281 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5341 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5401 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5461 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5521 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5581 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5641 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5701 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5761 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5821 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5881 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5941 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6001 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6061 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6121 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6181 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6241 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6301 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6421 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6481 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6541 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6601 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6661 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6721 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6781 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6841 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6901 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6961 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7021 cctggccttt tgctggcctt ttgctcacat gt
//
