pAAV-hSyn-SOUL-P2A-tdTomato

LOCUS       pAAV-hSyn-SOUL-P2A-tdTomato                7050 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Step-function opsin with ultra-high light sensitivity (SOUL). 
            Capable to be used for noninvasive stimulation in mice and minimally
            invasive stimulation in macaque..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-SOUL-P2A-tdTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7050)
  AUTHORS   Gong X, Mendoza-Halliday D, Ting JT, Kaiser T, Sun X, Bastos AM, 
            Wimmer RD, Guo B, Chen Q, Zhou Y, Pruner M, Wu CW, Park D, 
            Deisseroth K, Barak B, Boyden ES, Miller EK, Halassa MM, Fu Z, Bi G,
            Desimone R, Feng G
  TITLE     An Ultra-Sensitive Step-Function Opsin for Minimally Invasive 
            Optogenetic Stimulation in Mice and Macaques.
  JOURNAL   Neuron. 2020 Apr 14. pii: S0896-6273(20)30239-7. doi: 
            10.1016/j.neuron.2020.03.032.
  PUBMED    32353253
REFERENCE   2  (bases 1 to 7050)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7050
                     /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        183..630
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      678..687
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             684..1610
                     /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
                     PSMLYLATGHRVQWLRYAEWLLTSPVILIHLSNLTGLSNDYSRRTMGLLVSAIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             1629..1685
                     /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             1686..3116
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    3177..3765
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3230..3250)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3648..3659)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3797..4273
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3996..4015)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4313..4453
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4313..4442
                     /label=AAV2 ITR
     rep_origin      4528..4983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4615..4634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4825..4846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5000..5019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5119..5141
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5179..5197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5265..5369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5370..6230
                     /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(5588..5607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6401..6989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6890..6909
                     /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 gcgtttaatt aagtgtctag actgcagagg gccctgcgta
      181 tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc
      241 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca
      301 gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg
      361 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc
      421 gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc
      481 cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg
      541 cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc
      601 ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggctagagga tccttcgaaa
      661 ccggtgctag cgcggccgcc accatggact atggcggcgc tttgtctgcc gtcggacgcg
      721 aacttttgtt cgttactaat cctgtggtgg tgaacgggtc cgtcctggtc cctgaggatc
      781 aatgttactg tgccggatgg attgaatctc gcggcacgaa cggcgctcag accgcgtcaa
      841 atgtcctgca gtggcttgca gcaggattca gcattttgct gctgatgttc tatgcctacc
      901 aaacctggaa atctacatgc ggctgggagg agatctatgt gtgcgccatt gaaatggtta
      961 aggtgattct cgagttcttt tttgagttta agaatccctc tatgctctac cttgccacag
     1021 gacaccgggt gcagtggctg cgctatgcag agtggctgct cacttctcct gtcatcctta
     1081 tccacctgag caacctcacc ggcctgagca acgactacag caggagaacc atgggactcc
     1141 ttgtctcagc catcgggtgt atcgtgtggg gggctaccag cgccatggca accggctatg
     1201 ttaaagtcat cttcttttgt cttggattgt gctatggcgc gaacacattt tttcacgccg
     1261 ccaaagcata tatcgagggt tatcatactg tgccaaaggg tcggtgccgc caggtcgtga
     1321 ccggcatggc atggctgttt ttcgtgagct ggggtatgtt cccaattctc ttcattttgg
     1381 ggcccgaagg ttttggcgtc ctgagcgtct atggctccac cgtaggtcac acgattattg
     1441 atctgatgag taaaaattgt tgggggttgt tgggacacta cctgcgcgtc ctgatccacg
     1501 agcacatatt gattcacgga gatatccgca aaaccaccaa actgaacatc ggcggaacgg
     1561 agatcgaggt cgagactctc gtcgaagacg aagccgaggc cggagccgtg ccagttaacg
     1621 gaagcggagc cactaacttc tccctgttga aacaagcagg ggatgtcgaa gagaatcccg
     1681 ggccaatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca
     1741 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1801 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     1861 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
     1921 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1981 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
     2041 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     2101 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     2161 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     2221 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
     2281 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     2341 agggccgcca ccacctgttc ctggggcatg gcatcggcag caccggcagc ggcagctccg
     2401 gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg cgcttcaagg
     2461 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     2521 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     2581 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
     2641 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
     2701 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg
     2761 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
     2821 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
     2881 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg
     2941 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
     3001 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     3061 gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagtaaggcg
     3121 cgaatcgcga gtcgacggcg cgcccctgca gggaattcga tatcaagctt atcgataatc
     3181 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3241 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3301 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3361 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3421 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3481 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3541 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3601 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3661 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3721 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     3781 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3841 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3901 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3961 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     4021 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     4081 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     4141 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     4201 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     4261 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     4321 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4381 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4441 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4501 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4561 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4621 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4681 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4741 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4801 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     4861 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     4921 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4981 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5041 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5101 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5161 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5221 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5281 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5341 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5401 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5461 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5521 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5581 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5641 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5701 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5761 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5821 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5881 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5941 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6001 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6061 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6121 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6181 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6241 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6301 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6541 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7021 tggccttttg ctggcctttt gctcacatgt
//