LOCUS       Exported                4247 bp ds-DNA     linear   SYN 20-MAY-2021
DEFINITION  Cation channelrhodopsin ChroME targeted to the neuronal soma and 
            proximal dendrites and separated from nuclear mRuby3 by P2A in a 
            viral vector.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CAG-DIO-ChroME-ST-P2A-H2B-mRuby3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4247)
  AUTHORS   Mardinly AR, Oldenburg IA, Pegard NC, Sridharan S, Lyall EH, Chesnov
            K, Brohawn SG, Waller L, Adesnik H
  TITLE     Precise multimodal optical control of neural ensemble activity.
  JOURNAL   Nat Neurosci. 2018 Apr 30. pii: 10.1038/s41593-018-0139-8. doi: 
            10.1038/s41593-018-0139-8.
  PUBMED    29713079
REFERENCE   2  (bases 1 to 4247)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 20, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4247
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     378..400
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     462..481
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    532..565
                     /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    616..649
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(661..1371)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     CDS             complement(1381..1758)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             complement(1765..1821)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(2032..2055)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(3055..3088)
                     /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(3139..3172)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3197..3785
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3250..3270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3668..3679)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3788..3804)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3789..3805)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3834..3955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3845..3864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3899..3918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4117..4246
                     /label=AAV2 ITR
ORIGIN
        1 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
       61 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      121 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      181 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      241 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      301 ggccctttgt gcggggggag cggctcgggg ctgtccgcgg ggggacggct gccttcgggg
      361 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
      421 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
      481 gctgtctcat cattttggca aagaattgga tcctctagag tcgactccgg aataacttcg
      541 tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca
      601 atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg gcgcgcctca
      661 cttgtacagc tcgtccatgc caccaccaag gttgctgtat ttggcaactg ccacttctct
      721 ttgcactacg taggtttcat tgtcactctc ctcgatcctt tccaggcggt gatcaacggc
      781 atggacaccg ggcatcttga tgttcccgac ggtctttttt gacctgtaag ttgtcacgaa
      841 gttgcagtgc agatggccac caccatcaac tttcagtgcg atgtcagtgt atcctctcag
      901 accaccatct gctggataca tcatctctgt attaggctcc caacccttgg tcttcttctg
      961 catcacggga ccattggagg gaaagtttac ccctctgacc ttgacgttgt agacgagctc
     1021 gccatcctca aggctggtgt cctgcgtgac ggtgacgact ccaccatctt cgtatctcgt
     1081 aactctttcc caagtaaaac cctcaggaaa ggactgttta aagaaatcag ggatgtcggc
     1141 cgggtacttg ataaaggtac ggctgccata catgaacgac gtggcaagaa tgtcaaaggc
     1201 aaatggcagg ggtcctccct cgatgacttt gatcctcatg gtttgcactc cctcgtacgg
     1261 tctgccttct ccttcacctg tgcatttgaa ttggtggccg ttgaccgaac cttccatgac
     1321 caccttcata cgcatatttt ccttgatcag ctcttcgccc ttagacacca tctcgagtgc
     1381 cttagcgctg gtgtacttgg tgatggcctt agtaccctcg gacacggcgt gcttggccaa
     1441 ctccccaggc agcagcaggc gcacggccgt ctggatctcc ctggaggtga tggtcgagcg
     1501 cttgttgtaa tgcgccaggc gggaagcctc acctgcgatg cgctcgaaaa tgtcgttcac
     1561 aaacgaattc atgatgccca tggccttgga cgaaatgccg gtgtcagggt ggacctgctt
     1621 cagaaccttg tacacataga tggaatagct ctccttgcgg ctgcgcttgc gcttcttgcc
     1681 gcctttcttc tgcgccttag tcaccgcctt cttggagccc tttttcgggg cgggagcaga
     1741 cttcgctggc tctggcatac tagtaggtcc agggttctcc tccacgtctc cagcctgctt
     1801 cagcaggctg aagttagtag ctccgcttcc tgccgcgaat ctggtggctt cagggaagtc
     1861 cgtggcacag ctgatgaagc tgtcaatgct ggacatgctc cgcatgtcga tgacgccctc
     1921 cgtgcgtgcg ggcagagggg ccacggggct gggcatgctg ctcatctcca gctcctctgg
     1981 aggcttgctc tgcggggcca tctccttggt gttgaggatg ggctgggact gcttatcgtc
     2041 atcgtctttg taatctaccg gtgcggccgc cactcctccc tcctcctcct cgtcgacgaa
     2101 ggtctcgatc tccatgttct cgccggccac gttgatggtg gttgtcttcc ggatgtcgcc
     2161 gtggatcagg atgtgctcgt ggatcttcac tctcaggtgg tggcccagga agccccaagt
     2221 gttcttgctg atcagatcca ggatcaggtg gccgattccg ctggtgtaag gtgtgatcag
     2281 gcccagtccc tcgtgtccgg cgatgaacat cacggggaac atcagccaag agcagaagaa
     2341 gacgtaggcc atgatcttgc aaatcttccg gcagacgcct ttgggcaggg tgtggtagct
     2401 ctcgatatac accttggcga tctggaagaa gcaagtcacg ccgtagaaca ggccgatcat
     2461 gaagaacagg atcttcaggg ggccctttgt aaaggcggct gtgatccccc acacgatgtt
     2521 gccgatgtcg gtcaccagga tggtcatggt ccgcttgctg tactcttcgt gcagtccggt
     2581 caggttgctc aggtggatca ggatcacggg gcaagtcagg agccactcgc tgtaccgcag
     2641 ccaaatcacg gctcctccgt tggtctggta cactgtggct gggctgtcga cctcgtggaa
     2701 cagctcgatg aagcacttga cgagctcgat cacgcagacg tagacctcct cccagccggt
     2761 tgtggccttg aagctgtgcc aaccgtagaa gatgagctgc acgatagcca cgatcacggt
     2821 gaaccactgg agtcccactg cgatcttgtg ctgcagctcg gttccgtggt tgatgtgggg
     2881 gaagcaaccg tgatcggctc cggctgttgt ggcgttagag gtttctccgt gagcgtctgc
     2941 tgctggtgtc accactgctg cggcgctcag caggcctcta attgtggctt cggcgctagg
     3001 cacggctgag ataaaggcgt gggtcattgt ggcggctgtt tccatactgc tagcataact
     3061 tcgtataaag tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc
     3121 tttagaatgg tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga
     3181 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3241 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3301 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3361 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3421 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3481 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3541 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3601 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3661 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3721 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3781 cccgcatcga taccgtcgac ctcgacccgg gcggccgctt cgagcagaca tgataagata
     3841 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3901 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3961 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     4021 caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg
     4081 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     4141 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4201 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagc
//
