LOCUS       Exported                7947 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  High efficiency channelrhodopsin, neuron specific promoter, red 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV hSyn1 ChR2 ET-TC 2A tDimer
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7947)
  AUTHORS   Berndt A, Schoenenberger P, Mattis J, Tye KM, Deisseroth K, Hegemann
            P, Oertner TG
  TITLE     High-efficiency channelrhodopsins for fast neuronal stimulation at 
            low light levels.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 May 3;108(18):7595-600. doi: 
            10.1073/pnas.1017210108. Epub 2011 Apr 19.
  PUBMED    21504945
REFERENCE   2  (bases 1 to 7947)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7947
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      466..475
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             472..1395
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYATWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGA"
     CDS             1408..1461
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      2413..2422
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      2417..2426
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    3841..4429
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3894..3914)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4312..4323)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4432..4448)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4433..4449)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4516..4992
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4715..4734)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5032..5172
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5032..5161
                     /label=AAV2 ITR
     rep_origin      5247..5702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5334..5353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5544..5565
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5719..5738)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5838..5860
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5898..5916)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5984..6088
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6089..6949
                     /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(6307..6326)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7120..7708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7609..7628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7770..7899
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gaattcaagc tgctagccac catggattat
      481 ggaggcgccc tgagtgccgt tgggcgcgag ctgctatttg taacgaaccc agtagtcgtc
      541 aatggctctg tacttgtgcc tgaggaccag tgttactgcg cgggctggat tgagtcgcgt
      601 ggcacaaacg gtgcccaaac ggcgtcgaac gtgctgcaat ggcttgctgc tggcttctcc
      661 atcctactgc ttatgtttta cgcctaccaa acatggaagt caacctgcgg ctgggaggag
      721 atctatgtgt gcgctatcga gatggtcaag gtgattctcg agttcttctt cgagtttaag
      781 aacccgtcca tgctgtatct agccacaggc caccgcgtcc agtggttgcg ttacgccacg
      841 tggcttctca cctgcccggt cattctcatt cacctgtcaa acctgacggg cttgtccaac
      901 gactacagca ggcgcaccat gggtctgctt gtgtctgata ttggctgtat tgtgtggggc
      961 gccacttccg ccatggccac cggatacgtc aaggtcatct tcttctgcct gggtctgtgt
     1021 tatggtgcta acacgttctt tcacgctgcc aaggcctaca tcgagggtta ccacaccgtg
     1081 ccgaagggcc ggtgtcgcca ggtggtgact ggcatggctt ggctcttctt cgtatcatgg
     1141 ggtatgttcc ccatcctgtt catcctcggc cccgagggct tcggcgtcct gagcgtgtac
     1201 ggctccaccg tcggccacac catcattgac ctgatgtcga agaactgctg gggtctgctc
     1261 ggccactacc tgcgcgtgct gatccacgag catatcctca tccacggcga cattcgcaag
     1321 accaccaaat tgaacattgg tggcactgag attgaggtcg agacgctggt ggaggacgag
     1381 gccgaggctg gcgcggtacc cgcggccgag ggcagaggaa gtcttctaac atgcggtgac
     1441 gtggaggaga atcccggccc tgcaccggga tccggcatgg acgtggtgaa tcagctggtg
     1501 gctgggggtc agttccgggt ggtcaaggag ccccttggct tcgtgaaggt gctgcagtgg
     1561 gtctttgcca tcttcgcctt tgctacgtgt ggcagctaca ccggggagct tcggctgagc
     1621 gtggagtgtg ccaacaagac ggagagtgcc ctcaacatcg aagttgaatt cgagtacccc
     1681 ttcaggctgc accaagtgta ctttgatgca ccctcctgcg tcaaaggggg cactaccaag
     1741 atcttcctgg ttggggacta ctcctcgtcg gctgaattct ttgtcaccgt ggctgtgttt
     1801 gccttcctct actccatggg ggccctggcc acctacatct tcctgcagaa caagtaccga
     1861 gagaacaaca aagggcctat gatggacttt ctggctacag ccgtgttcgc tttcatgtgg
     1921 ctagttagtt catcagcctg ggccaaaggc ctgtccgatg tgaagatggc cacggaccca
     1981 gagaacatta tcaaggagat gcccatgtgc cgccagacag ggaacacatg caaggaactg
     2041 agggaccctg tgacttcagg actcaacacc tcagtggtgt ttggcttcct gaacctggtg
     2101 ctctgggttg gcaacttatg gttcgtgttc aaggagacag gctgggcagc cccattcatg
     2161 cgcgcacctc caggcgcccc ggaaaagcaa ccagcacctg gcgatgccta cggcgatgcg
     2221 ggctacgggc agggccccgg aggctatggg ccccaggact cctacgggcc tcagggtggt
     2281 tatcaacccg attacgggca gccagccagc ggtggcggtg gctacgggcc tcagggcgac
     2341 tatgggcagc aaggctatgg ccaacagggt gcgcccacct ccttctccaa tcagatggcg
     2401 gccggatccg cggccgccat ggtggcctcc tccgaggacg tcatcaaaga gttcatgcgc
     2461 ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2521 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     2581 ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa ggcgtacgtg
     2641 aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg
     2701 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2761 caggacggca cgctgatcta caaggtgaag ttccgcggca ccaacttccc ccccgacggc
     2821 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     2881 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     2941 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcagctgcc cggctactac
     3001 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     3061 tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggcaccgg cagcaccggc
     3121 agcggcagct ccggcaccgc ctcctccgag gacgtcatca aagagttcat gcgcttcaag
     3181 gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     3241 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     3301 ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggcgta cgtgaagcac
     3361 cccgccgaca tccccgacta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     3421 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     3481 ggcacgctga tctacaaggt gaagttccgc ggcaccaact tcccccccga cggccccgta
     3541 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     3601 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     3661 gagttcaaga ccatctacat ggccaagaag cccgtgcagc tgcccggcta ctactacgtg
     3721 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     3781 cgctccgagg gccgccacca cctgttcctg taggtcgact tcgagcaaaa gcttatcgat
     3841 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3901 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3961 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4021 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4081 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4141 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4201 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     4261 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4321 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4381 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     4441 cgacctcgag acgcgatcgc ggcgcgccat tcgatggcct ctctggcctc gacctcgagt
     4501 ctagagatga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     4561 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     4621 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     4681 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     4741 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     4801 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     4861 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     4921 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     4981 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     5041 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     5101 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     5161 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5221 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     5281 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     5341 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5401 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5461 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5521 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     5581 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     5641 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5701 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5761 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5821 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5881 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5941 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6001 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6061 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6121 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6181 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6241 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6301 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6361 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6421 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6481 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6541 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6601 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6661 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6721 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6781 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6841 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6901 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6961 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7021 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7081 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7141 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7201 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7261 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7321 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7381 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7441 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7501 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7561 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7621 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7681 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7741 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
     7801 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     7861 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgtcgctaga
     7921 ctcgagctgc agagggccct gcgtatg
//
