LOCUS       Exported                7889 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-BiPOLES-mCerulean
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7889)
  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 7889)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7889
                     /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(789..1835)
                     /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(2235..2256)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2584..2603
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2843..2864
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3874..3907)
                     /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(3958..3991)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4016..4604
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4069..4089)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4487..4498)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4636..5112
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4835..4854)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5152..5292
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5152..5281
                     /label=AAV2 ITR
     rep_origin      5367..5822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5454..5473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5664..5685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5839..5858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5958..5980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6018..6036)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6104..6208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6209..7069
                     /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(6427..6446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7240..7828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7729..7748
                     /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 acactgtgtc ctcgtcctcc tcctccacga actcttccac ttccacctcc
      841 tcgcctccga tctccatctt ggtggtcttc cggatgtcgc cgtggatcag gatgtgctcg
      901 tggatcttga tcctcaggtg gtgggccagg aaggtccaaa actccttggc gatgatgtcg
      961 cagatgctgt ggccgatgct gttggcgtaa gggctcagct tcagcagtcc ttctggtccc
     1021 actgcccaga ggattgggta gctgccccaa gaggcgaagt aagcgtaggc catcagcttc
     1081 acgaccatgc ggcaatggcc tttaggcacg ctgtggttgg cttccacgta gcacttggcg
     1141 gcctggaagt acatgtagcc gccgtagatg caagacacga tatacagcag ccacttgagc
     1201 caatcggtag ccagtcctgc ggccatgccg aacacgatca ttcccacgca agacacgatc
     1261 aggcccatgg tccgcttgct gtagtcgttc ttcaggccgc tcaggttgct cagcttgatc
     1321 aggatcacgg ggcaagacag cagccactcg aagtagcgca ggcaataggc gtggttgccg
     1381 gtagacaggt acactgtggc ggggctgctg aactccttga agatctccag ggtcacgaac
     1441 agcacctcga cgcaacagac gtagacctcc tcccaaccgc aagtggcctt ccaagcgctg
     1501 aagccgtaga atgtcagcag ggcgatggcg atgctgaaag caatccactg gcagacctgg
     1561 gcgccgatct tttctcctgg tgtgcctctg ctgtgcagcc aagcctgctt cttctccact
     1621 cccacgacca ggtagccgcc tgtggcctcc acgaagcaat atccgtatcc ggcgtcgctc
     1681 agtccgtaag aagggtcaca ccaccactcg gtgctgaagc actcgcctgt aggtgtcatt
     1741 cctccacagc ccatgtcctc gtggtggtag gggttaggcc aagggttgat gcctccggcg
     1801 gcaaacagag atctggtggc gctgctgatc agctcttgta cagctccctc cggacgtccg
     1861 gtgatctggg cctgtctaga gctgttttca gagatgttgt aggaaatctg cagccctctt
     1921 tccccataca catccggtct caaggttacc cccggtgact ttaactggtc ctggtagtcg
     1981 ggggtgtagg gatcaatggt ctgcatcagc acatagatgc acaaggcaaa gagcccagtc
     2041 atgaccacgt agaaagctgc atagtacagg ctgatccaca cccaccgggc tggggtgcgg
     2101 cccagcatct gcccagtgtc cgggttccaa cagtattgcc ggaattcggc catgcgctgg
     2161 ctgcatgact tcttctcctg cagggctgca ccggtataca gctcgtccat gccgagagtg
     2221 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     2281 ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     2341 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     2401 cagttgaggc cgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
     2461 ttgccgtgga tggcgttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     2521 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     2581 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     2641 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     2701 tgcacgcccc agctcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     2761 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     2821 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     2881 gtgaacagct cctcgccctt gctcacgggt accgccacgt tgatgtcgat ctggtccagg
     2941 gggatgtact cgccctcgct ggtgatcctg ctcttggccg ccgtacgtcc ccttccaaac
     3001 attctgatat tgacttcgtc gttaggaggg gcattcttct cgtgattcag gccaaaagga
     3061 tcgtcggctt cgggtttctc ctgccctggc agacatgcgg ggtcccactt cccttccagc
     3121 actccccagg cagtgctcca caggatcact ccatatgtat ttttgcacag gccgtcagcc
     3181 agcaggtaca gaatggcgct cacgttctcg tggatcacgc agactccggt gggtgaaatc
     3241 agccacacga ttgggaacat agtccaagca ataaagaaga tcaggcgcag catcttgatc
     3301 ctcaccacca ccttctgtgc cagaggtgtc ccgacagatt caaagccata cagccccact
     3361 tgaaagattg tgaagaccac actgtactcg aacaccaggc aggtggcccc aaagaaaaag
     3421 aacagccact tcatatactc tgcgggagaa attgtagcgg tgattcccat gaccactcta
     3481 atgatggaag cggcctgtgc aatgggattc agtgggatac ttttgtactt caccagagcc
     3541 atgttagaaa tctgtcccag catgattgga caggtgcaca gccaggaggc cattcggctc
     3601 caaggaaaga atctgccgtc cctcatctgc agggttccgt tgccagtact ggccagtgca
     3661 taagtgatag attcgacaaa gggcaggtac acggactccc aagggacctt gggagccctg
     3721 ctaatcatga agattgtctg gcagcctgag accacggccc acaccagctg cagcagtgag
     3781 acgaaggtgg agtcgattct ggacatattg tagcactcgg tgtgagttga tgcccattcc
     3841 ccatagacga cctgtgatgc catggatcct agcataactt cgtataaagt atcctatacg
     3901 aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata
     3961 acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca
     4021 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4081 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4141 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4201 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4261 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4321 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4381 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
     4441 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4501 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4561 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
     4621 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     4681 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     4741 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     4801 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     4861 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     4921 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     4981 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     5041 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     5101 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     5161 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     5221 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     5281 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5341 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     5401 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     5461 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5521 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5581 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5641 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     5701 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     5761 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5821 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5881 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5941 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6001 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6061 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6421 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6721 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7381 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7861 ggccttttgc tggccttttg ctcacatgt
//
