LOCUS       Exported                6884 bp ds-DNA     linear   SYN 07-JUN-2023
DEFINITION  AAV expression of humanized ChR2 with H134R mutation fused to 
            mCherry driven by CAG promoter for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CAG.hChR2(H134R)-mCherry.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6884)
  TITLE     Deisseroth AAV plasmids
REFERENCE   2  (bases 1 to 6884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 7, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6884
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     406..428
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     490..509
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      579..588
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             585..1511
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             1524..2231
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1666..1684)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1934..1953
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2257..2389
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2372..2391
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2445..3033
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2498..2518)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2916..2927)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3076..3197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3141..3160
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3359..3488
                     /label=AAV2 ITR
     primer_bind     complement(3507..3524)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3507..3523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3516..3538)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3665..4120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3752..3771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3962..3983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4146..4250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4251..5111
                     /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(4469..4488)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5282..5870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5771..5790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6024..6041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6158..6179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6194..6224
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6232..6248
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6237..6259
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6256..6272
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6256..6272
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
       61 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      121 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      181 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      241 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      301 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcggggct
      361 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      421 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      481 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc
      541 gggatccact agtccagtgt ggtggaattg cccttgctgc caccatggac tatggcggcg
      601 ctttgtctgc cgtcggacgc gaacttttgt tcgttactaa tcctgtggtg gtgaacgggt
      661 ccgtcctggt ccctgaggat caatgttact gtgccggatg gattgaatct cgcggcacga
      721 acggcgctca gaccgcgtca aatgtcctgc agtggcttgc agcaggattc agcattttgc
      781 tgctgatgtt ctatgcctac caaacctgga aatctacatg cggctgggag gagatctatg
      841 tgtgcgccat tgaaatggtt aaggtgattc tcgagttctt ttttgagttt aagaatccct
      901 ctatgctcta ccttgccaca ggacaccggg tgcagtggct gcgctatgca gagtggctgc
      961 tcacttgtcc tgtcatcctt atccgcctga gcaacctcac cggcctgagc aacgactaca
     1021 gcaggagaac catgggactc cttgtctcag acatcgggac tatcgtgtgg ggggctacca
     1081 gcgccatggc aaccggctat gttaaagtca tcttcttttg tcttggattg tgctatggcg
     1141 cgaacacatt ttttcacgcc gccaaagcat atatcgaggg ttatcatact gtgccaaagg
     1201 gtcggtgccg ccaggtcgtg accggcatgg catggctgtt tttcgtgagc tggggtatgt
     1261 tcccaattct cttcattttg gggcccgaag gttttggcgt cctgagcgtc tatggctcca
     1321 ccgtaggtca cacgattatt gatctgatga gtaaaaattg ttgggggttg ttgggacact
     1381 acctgcgcgt cctgatccac gagcacatat tgattcacgg agatatccgc aaaaccacca
     1441 aactgaacat cggcggaacg gagatcgagg tcgagactct cgtcgaagac gaagccgagg
     1501 ccggagccgt gccagcggcc gccgtgagca agggcgagga ggataacatg gccatcatca
     1561 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     1621 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1681 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     1741 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     1801 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     1861 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     1921 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     1981 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     2041 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2101 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     2161 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2221 tgtacaagta aagcggcccc ggactcgagg ccgcaggtaa gtatcaaggt tacaagacag
     2281 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
     2341 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgtcgagtgg
     2401 agctcgcgac tagtcgattc gaattcgata tcaagcttat cgataatcaa cctctggatt
     2461 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2521 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2581 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2641 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2701 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2761 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2821 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     2881 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2941 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3001 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc
     3061 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3121 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3181 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3241 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3301 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     3361 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3421 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3481 gcgcgcagcc ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa
     3541 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3601 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3661 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3721 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3781 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3841 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3901 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3961 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4021 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4081 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     4141 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4201 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4261 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4321 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4381 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4441 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4501 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4561 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4621 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4681 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4741 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4801 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4861 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4921 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4981 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5041 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5101 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5161 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5221 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5281 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5341 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5401 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5461 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5521 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5581 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5641 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5701 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5761 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5821 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5881 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5941 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6001 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     6061 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     6121 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     6181 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     6241 gataacaatt tcacacagga aacagctatg accatgatta cgccagattt aattaagggc
     6301 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc
     6361 tcgctcactg aggccgcccg ggcaaagccc gggcggcctc agtgagcgag cgagcgcgca
     6421 gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat
     6481 gctacttatc tacgtagcca tgctctagga agagtaccat tgacgtcaat aatgacgtat
     6541 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6601 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     6661 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     6721 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc
     6781 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
     6841 attttttaat tattttgtgc agcgatgggg gcgggggggg gggg
//
