LOCUS       Exported                8095 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Optogenetic tool for blue-light inhibition and red-light excitation 
            of neurons.
ACCESSION   .
VERSION     .
KEYWORDS    hSyn-DIO-somBiPOLES-mCerulean
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8095)
  AUTHORS   Vierock J, Rodriguez-Rozada S, Pieper F, Dieter A, Bergs A, 
            Zeitzschel N, Ahlbeck J, Sauter K, Gottschalk A, Engel AK, Hegemann 
            P, Wiegert JS
  TITLE     BiPOLES: a tool for bidirectional dual-color optogenetic control of 
            neurons
  JOURNAL   bioRxiv (2020)
REFERENCE   2  (bases 1 to 8095)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8095
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    660..693
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    744..777
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(999..2042)
                     /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(2442..2463)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2791..2810
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3050..3071
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(4080..4113)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(4164..4197)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4222..4810
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4275..4295)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4693..4704)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4842..5318
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5041..5060)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5358..5498
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5358..5487
                     /label=AAV2 ITR
     rep_origin      5573..6028
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5660..5679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5870..5891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6045..6064)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6164..6186
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6224..6242)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6310..6414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6415..7275
                     /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(6633..6652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7446..8034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7935..7954
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgagctttc tagatcagaa gcgggtggcc tcggggaagt cggtggcgca gctgatgaag
      841 ctgtcgatgc tgctcatgct gcgcatgtcg atcacgccct cggtgcgggc gggcaggggg
      901 gccacggggc tgggcatgct gctcatctcc agctcctcgg ggggcttgct ctggggggcc
      961 atctccttgg tgttcaggat gggctggctc tgctcgagca ctgtgtcctc gtcctcctcc
     1021 tccacgaact cttccacttc cacctcctcg cctccgatct ccatcttggt ggtcttccgg
     1081 atgtcgccgt ggatcaggat gtgctcgtgg atcttgatcc tcaggtggtg ggccaggaag
     1141 gtccaaaact ccttggcgat gatgtcgcag atgctgtggc cgatgctgtt ggcgtaaggg
     1201 ctcagcttca gcagtccttc tggtcccact gcccagagga ttgggtagct gccccaagag
     1261 gcgaagtaag cgtaggccat cagcttcacg accatgcggc aatggccttt aggcacgctg
     1321 tggttggctt ccacgtagca cttggcggcc tggaagtaca tgtagccgcc gtagatgcaa
     1381 gacacgatat acagcagcca cttgagccaa tcggtagcca gtcctgcggc catgccgaac
     1441 acgatcattc ccacgcaaga cacgatcagg cccatggtcc gcttgctgta gtcgttcttc
     1501 aggccgctca ggttgctcag cttgatcagg atcacggggc aagacagcag ccactcgaag
     1561 tagcgcaggc aataggcgtg gttgccggta gacaggtaca ctgtggcggg gctgctgaac
     1621 tccttgaaga tctccagggt cacgaacagc acctcgacgc aacagacgta gacctcctcc
     1681 caaccgcaag tggccttcca agcgctgaag ccgtagaatg tcagcagggc gatggcgatg
     1741 ctgaaagcaa tccactggca gacctgggcg ccgatctttt ctcctggtgt gcctctgctg
     1801 tgcagccaag cctgcttctt ctccactccc acgaccaggt agccgcctgt ggcctccacg
     1861 aagcaatatc cgtatccggc gtcgctcagt ccgtaagaag ggtcacacca ccactcggtg
     1921 ctgaagcact cgcctgtagg tgtcattcct ccacagccca tgtcctcgtg gtggtagggg
     1981 ttaggccaag ggttgatgcc tccggcggca aacagagatc tggtggcgct gctgatcagc
     2041 tcttgtacag ctccctccgg acgtccggtg atctgggcct gtctagagct gttttcagag
     2101 atgttgtagg aaatctgcag ccctctttcc ccatacacat ccggtctcaa ggttaccccc
     2161 ggtgacttta actggtcctg gtagtcgggg gtgtagggat caatggtctg catcagcaca
     2221 tagatgcaca aggcaaagag cccagtcatg accacgtaga aagctgcata gtacaggctg
     2281 atccacaccc accgggctgg ggtgcggccc agcatctgcc cagtgtccgg gttccaacag
     2341 tattgccgga attcggccat gcgctggctg catgacttct tctcctgcag ggctgcaccg
     2401 gtatacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     2461 atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct caggtagtgg
     2521 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     2581 agctgcacgc tgccgtcctc gatgttgcag ttgaggccga agttggcctt gatgccgttc
     2641 ttctgcttgt cggcggtgat atagacgttg ccgtggatgg cgttgtactc cagcttgtgc
     2701 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     2761 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     2821 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     2881 atgtggtcgg ggtagcgggc gaagcactgc acgccccagc tcagggtggt cacgagggtg
     2941 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     3001 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     3061 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct cacgggtacc
     3121 gccacgttga tgtcgatctg gtccaggggg atgtactcgc cctcgctggt gatcctgctc
     3181 ttggccgccg tacgtcccct tccaaacatt ctgatattga cttcgtcgtt aggaggggca
     3241 ttcttctcgt gattcaggcc aaaaggatcg tcggcttcgg gtttctcctg ccctggcaga
     3301 catgcggggt cccacttccc ttccagcact ccccaggcag tgctccacag gatcactcca
     3361 tatgtatttt tgcacaggcc gtcagccagc aggtacagaa tggcgctcac gttctcgtgg
     3421 atcacgcaga ctccggtggg tgaaatcagc cacacgattg ggaacatagt ccaagcaata
     3481 aagaagatca ggcgcagcat cttgatcctc accaccacct tctgtgccag aggtgtcccg
     3541 acagattcaa agccatacag ccccacttga aagattgtga agaccacact gtactcgaac
     3601 accaggcagg tggccccaaa gaaaaagaac agccacttca tatactctgc gggagaaatt
     3661 gtagcggtga ttcccatgac cactctaatg atggaagcgg cctgtgcaat gggattcagt
     3721 gggatacttt tgtacttcac cagagccatg ttagaaatct gtcccagcat gattggacag
     3781 gtgcacagcc aggaggccat tcggctccaa ggaaagaatc tgccgtccct catctgcagg
     3841 gttccgttgc cagtactggc cagtgcataa gtgatagatt cgacaaaggg caggtacacg
     3901 gactcccaag ggaccttggg agccctgcta atcatgaaga ttgtctggca gcctgagacc
     3961 acggcccaca ccagctgcag cagtgagacg aaggtggagt cgattctgga catattgtag
     4021 cactcggtgt gagttgatgc ccattcccca tagacgacct gtgatgccat ggatctagca
     4081 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt
     4141 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa
     4201 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     4261 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4321 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4381 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4441 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4501 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4561 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4621 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     4681 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4741 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4801 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     4861 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     4921 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     4981 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     5041 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     5101 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     5161 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     5221 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     5281 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     5341 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     5401 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     5461 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     5521 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     5581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5641 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5701 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5761 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     5821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5881 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     5941 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6001 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     6061 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     6121 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     6181 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     6241 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6301 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6361 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6421 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6481 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6541 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6601 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6661 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6721 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6781 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6841 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6901 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6961 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7021 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7081 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7141 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7201 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7261 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7321 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7381 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7441 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7501 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7561 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7621 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7681 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7741 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7801 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7861 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7921 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7981 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8041 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
