LOCUS       Exported                6944 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Channelrhodopsin localized to neuronal soma with simultaneous 
            nuclear labeling.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-syn-ChR2-Kv-P2A-H2b-mRuby2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   Baker CA, Elyada YM, Parra A, Bolton MM
  TITLE     Cellular resolution circuit mapping with temporal-focused excitation
            of soma-targeted channelrhodopsin.
  JOURNAL   Elife. 2016 Aug 15;5. pii: e14193. doi: 10.7554/eLife.14193.
  PUBMED    27525487
REFERENCE   2  (bases 1 to 6944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6944
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      671..680
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             677..1603
                     /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             1823..1879
                     /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             1886..2263
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      2327..2336
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2333..3043
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    3071..3659
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3124..3144)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3542..3553)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3691..4167
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3890..3909)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4207..4347
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4207..4336
                     /label=AAV2 ITR
     rep_origin      4422..4877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4509..4528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4719..4740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4894..4913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5013..5035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5073..5091)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5159..5263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5264..6124
                     /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(5482..5501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6295..6883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6784..6803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgactccgga gccaccatgg actatggcgg cgctttgtct gccgtcggac gcgaactttt
      721 gttcgttact aatcctgtgg tggtgaacgg gtccgtcctg gtccctgagg atcaatgtta
      781 ctgtgccgga tggattgaat ctcgcggcac gaacggcgct cagaccgcgt caaatgtcct
      841 gcagtggctt gcagcaggat tcagcatttt gctgctgatg ttctatgcct accaaacctg
      901 gaaatctaca tgcggctggg aggagatcta tgtgtgcgcc attgaaatgg ttaaggtgat
      961 tctcgagttc ttttttgagt ttaagaatcc ctctatgctc taccttgcca caggacaccg
     1021 ggtgcagtgg ctgcgctatg cagagtggct gctcacttgt cctgtcatcc ttatccgcct
     1081 gagcaacctc accggcctga gcaacgacta cagcaggaga accatgggac tccttgtctc
     1141 agacatcggg actatcgtgt ggggggctac cagcgccatg gcaaccggct atgttaaagt
     1201 catcttcttt tgtcttggat tgtgctatgg cgcgaacaca ttttttcacg ccgccaaagc
     1261 atatatcgag ggttatcata ctgtgccaaa gggtcggtgc cgccaggtcg tgaccggcat
     1321 ggcatggctg tttttcgtga gctggggtat gttcccaatt ctcttcattt tggggcccga
     1381 aggttttggc gtcctgagcg tctatggctc caccgtaggt cacacgatta ttgatctgat
     1441 gagtaaaaat tgttgggggt tgttgggaca ctacctgcgc gtcctgatcc acgagcacat
     1501 attgattcac ggagatatcc gcaaaaccac caaactgaac atcggcggaa cggagatcga
     1561 ggtcgagact ctcgtcgaag acgaagccga ggccggagcc gtgccagcgg ccgccaccca
     1621 gtcccagcct attctgaaca ccaaagagat ggccccccag agtaaacccc ctgaggagct
     1681 ggaaatgtcc tcaatgccct cccctgtggc acctcttcct gcaagaacag agggcgtgat
     1741 cgacatgagg tccatgagtt ctattgactc ttttatctca tgcgccaccg attttccaga
     1801 ggctacgcgg tttggatccg gagccacgaa cttctctctg ttaaagcaag caggagacgt
     1861 ggaagaaaac cccggtcctg gttctatgcc agagccagcg aagtctgctc ccgccccgaa
     1921 aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga agcgcaagcg
     1981 cagccgcaag gagagctatt ccatctatgt gtacaaggtt ctgaagcagg tccaccctga
     2041 caccggcatt tcgtccaagg ccatgggcat catgaattcg tttgtgaacg acattttcga
     2101 gcgcatcgca ggtgaggctt cccgcctggc gcattacaac aagcgctcga ccatcacctc
     2161 cagggagatc cagacggccg tgcgcctgct gctgcctggg gagttggcca agcacgccgt
     2221 gtccgagggt actaaggcca tcaccaagta caccagcgct aaggatccac cggtcgccac
     2281 catgctcgtc gaagacgaag ccgaggccgg agccgtgcca gcggccgcca ccatggtgtc
     2341 taagggcgaa gagctgatca aggaaaatat gcgtatgaag gtggtcatgg aaggttcggt
     2401 caacggccac caattcaaat gcacaggtga aggagaaggc aatccgtaca tgggaactca
     2461 aaccatgagg atcaaagtca tcgagggagg acccctgcca tttgcctttg acattcttgc
     2521 cacgtcgttc atgtatggca gccgtacttt tatcaagtac ccgaaaggca ttcctgattt
     2581 ctttaaacag tcctttcctg agggttttac ttgggaaaga gttacgagat acgaagatgg
     2641 tggagtcgtc accgtcatgc aggacaccag ccttgaggat ggctgtctcg tttaccacgt
     2701 ccaagtcaga ggggtaaact ttccctccaa tggtcccgtg atgcagaaga agaccaaggg
     2761 ttgggagcct aatacagaga tgatgtatcc agcagatggt ggtctgaggg gatacactca
     2821 tatggcactg aaagttgatg gtggtggcca tctgtcttgc tctttcgtaa caacttacag
     2881 gtcaaaaaag accgtcggga acatcaagat gcccggtatc catgccgttg atcaccgcct
     2941 ggaaaggtta gaggaaagtg acaatgaaat gttcgtagta caacgcgaac acgcagttgc
     3001 caagttcgcc gggcttggtg gtgggatgga cgagctgtac aagtaagaat tcgatatcaa
     3061 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3121 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3181 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3241 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3301 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3361 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3421 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3481 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3541 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3601 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3661 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3721 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3781 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3841 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3901 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3961 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     4021 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     4081 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     4141 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     4201 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4261 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4321 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4381 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4561 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4621 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4741 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4801 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4861 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4921 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4981 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5041 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     5101 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5281 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5341 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5401 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5461 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5521 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5581 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5641 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5701 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5761 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5821 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5881 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5941 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6001 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6061 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6121 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6181 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6241 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6421 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6901 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
