LOCUS       Exported                6545 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV-mediated expression of ChromeT-mCherry under the GFAP promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-GFAP104-ChromeT-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6545)
  AUTHORS   Cho YK, Park D, Yang A, Chen F, Chuong AS, Klapoetke NC, Boyden ES
  TITLE     Multidimensional screening yields channelrhodopsin variants having 
            improved photocurrent and order-of-magnitude reductions in calcium 
            and proton currents.
  JOURNAL   J Biol Chem. 2019 Jan 4. pii: RA118.006996. doi: 
            10.1074/jbc.RA118.006996.
  PUBMED    30610117
REFERENCE   2  (bases 1 to 6545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6545
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      998..1007
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1004..1930
                     /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
                     NGAQTASNVLQWLAAGFSILLLMFYSYQTWKSTCGWEEIYVCAIAMVKVILEFFFEFKN
                     PSMLYLATGGRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             1946..2653
                     /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(2088..2106)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2356..2375
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2678..3266
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2731..2751)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3149..3160)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3286..3762
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3485..3504)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3808..3948
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3808..3937
                     /label=AAV2 ITR
     rep_origin      4023..4478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4110..4129)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4320..4341
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4495..4514)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4614..4636
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4674..4692)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4760..4864
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4865..5725
                     /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(5083..5102)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5896..6484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6385..6404
                     /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 gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagcg
      301 ctagatctaa catatcctgg tgtggagtag gggacgctgc tctgacagag gctcgggggc
      361 ctgagctggc tctgtgagct ggggaggagg cagacagcca ggccttgtct gcaagcagac
      421 ctggcagcat tgggctggcc gccccccagg gcctcctctt catgcccagt gaatgactca
      481 ccttggcaca gacacaatgt tcggggtggg cacagtgcct gcttcccgcc gcaccccagc
      541 ccccctcaaa tgccttccga gaagcccatt gagcaggggg cttgcattgc accccagcct
      601 gacagcctgg catcttggga taaaagcagc acagccccct aggggctgcc cttgctgtgt
      661 ggcgccaccg gcggtggaga acaaggctct attcagcctg tgcccaggaa aggggatcag
      721 gggatgccca ggcatggaca gtgggtggca gggggggaga ggagggctgt ctgcttccca
      781 gaagtccaag gacacaaatg ggtgagggga gagctctccc catagctggg ctgcggccca
      841 accccacccc ctcaggctat gccagggggt gttgccaggg gcacccgggc atcgccagtc
      901 tagcccactc cttcataaag ccctcgcatc ccaggagcga gcagagccag agcaggttgg
      961 agaggagacg catcacctcc gctgctcgcg gggatccgcc accatggact atggcggcgc
     1021 tttgtctgcc gtcggacgcg aacttttgtt cgttactaat cctgtggtgg tgaacgggtc
     1081 cgtcctggtc cctgaggatc aatgttactg tgccggatgg attgaatctc gcggcacgaa
     1141 cggcgctcag accgcgtcaa atgtcctgca gtggcttgca gcaggattca gcattttgct
     1201 gctgatgttc tattcttacc aaacctggaa atctacatgc ggctgggagg agatctatgt
     1261 gtgcgccatt gcaatggtta aggtgattct cgagttcttt tttgagttta agaatccctc
     1321 tatgctctac cttgccacag gaggtcgggt gcagtggctg cgctatgcag agtggctgct
     1381 cacttgtcct gtcatcctta tccacctgag caacctcacc ggcctgagca acgactacag
     1441 caggagaacc atgggactcc ttgtctcaga catcgggact atcgtgtggg gggctaccag
     1501 cgccatggca accggctatg ttaaagtcat cttcttttgt cttggattgt gctatggcgc
     1561 gaacacattt tttcacgccg ccaaagcata tatcgagggt tatcatactg tgccaaaggg
     1621 tcggtgccgc caggtcgtga ccggcatggc atggctgttt ttcgtgagct ggggtatgtt
     1681 cccaattctc ttcattttgg ggcccgaagg ttttggcgtc ctgagcgtct atggctccac
     1741 cgtaggtcac acgattattg atctgatgag taaaaattgt tgggggttgt tgggacacta
     1801 cctgcgcgtc ctgatccacg agcacatatt gattcacgga gatatccgca aaaccaccaa
     1861 actgaacatc ggcggaacgg agatcgaggt cgagactctc gtcgaagacg aagccgaggc
     1921 cggagccgtg ccagcggcac cggtagtgag caagggcgag gaggataaca tggccatcat
     1981 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     2041 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2101 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     2161 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     2221 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     2281 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     2341 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     2401 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     2461 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     2521 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     2581 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     2641 gctgtacaag taagaattct gtacaaagct tatcgataat caacctctgg attacaaaat
     2701 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2761 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2821 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2881 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2941 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3001 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3061 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3121 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3181 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3241 gatctccctt tgggccgcct ccccgcatcg ataccggctc gagacgggtg gcatccctgt
     3301 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     3361 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     3421 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     3481 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     3541 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     3601 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     3661 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     3721 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctcgagct gattttgtag
     3781 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3841 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3901 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3961 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4021 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4081 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4141 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4201 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4261 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4321 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4381 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4441 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4501 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4561 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4621 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4681 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4741 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4801 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4861 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4921 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4981 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5041 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5101 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5161 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5221 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5281 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5341 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5401 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5461 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5521 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5581 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5641 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5701 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5761 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5821 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5881 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5941 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6001 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6061 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6541 catgt
//
