LOCUS pAAV-Syn-ChrimsonR-tdT 6864 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION AAV-mediated expression of ChrimsonR-tdTomato under the Syn
promoter.
ACCESSION .
VERSION .
KEYWORDS pAAV-Syn-ChrimsonR-tdT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6864)
AUTHORS Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK,
Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z,
Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V,
Constantine-Paton M, Wong GK, Boyden ES
TITLE Independent optical excitation of distinct neural populations.
JOURNAL Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub
2014 Feb 9.
PUBMED 24509633
REFERENCE 2 (bases 1 to 6864)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6864
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 142..161
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 320..449
/label=AAV2 ITR
promoter 549..996
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
regulatory 1009..1018
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1015..2064
/codon_start=1
/product="red light-drivable channelrhodopsin from
Chlamydomonas noctigama (Klapoetke et al., 2014)"
/label=ChrimsonR
/note="K176R mutant with accelerated closing kinetics"
/translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
FVEEEDEDTV"
misc_feature 3535..4123
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3588..3608)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4006..4017)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 4164..4371
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 4541..4670
/label=AAV2 ITR
primer_bind complement(4736..4753)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4907..5495)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4987..5006)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5666..6526)
/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 6289..6308
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6527..6631)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6657..248)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6794..6815)
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
61 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
121 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
181 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
241 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
301 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
361 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
421 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
481 atctacgtag ccatgctcta ggaagatcgt aggttaatta atctagactg cagagggccc
541 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
601 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
661 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
721 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
781 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
841 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
901 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
961 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga gaggatccgc caccatggct
1021 gagctgatca gcagcgccac cagatctctg tttgccgccg gaggcatcaa cccttggcct
1081 aacccctacc accacgagga catgggctgt ggaggaatga cacctacagg cgagtgcttc
1141 agcaccgagt ggtggtgtga cccttcttac ggactgagcg acgccggata cggatattgc
1201 ttcgtggagg ccacaggcgg ctacctggtc gtgggagtgg agaagaagca ggcttggctg
1261 cacagcagag gcacaccagg agaaaagatc ggcgcccagg tctgccagtg gattgctttc
1321 agcatcgcca tcgccctgct gacattctac ggcttcagcg cctggaaggc cacttgcggt
1381 tgggaggagg tctacgtctg ttgcgtcgag gtgctgttcg tgaccctgga gatcttcaag
1441 gagttcagca gccccgccac agtgtacctg tctaccggca accacgccta ttgcctgcgc
1501 tacttcgagt ggctgctgtc ttgccccgtg atcctgatca gactgagcaa cctgagcggc
1561 ctgaagaacg actacagcaa gcggaccatg ggcctgatcg tgtcttgcgt gggaatgatc
1621 gtgttcggca tggccgcagg actggctacc gattggctca agtggctgct gtatatcgtg
1681 tcttgcatct acggcggcta catgtacttc caggccgcca agtgctacgt ggaagccaac
1741 cacagcgtgc ctaaaggcca ttgccgcatg gtcgtgaagc tgatggccta cgcttacttc
1801 gcctcttggg gcagctaccc aatcctctgg gcagtgggac cagaaggact gctgaagctg
1861 agcccttacg ccaacagcat cggccacagc atctgcgaca tcatcgccaa ggagttttgg
1921 accttcctgg cccaccacct gaggatcaag atccacgagc acatcctgat ccacggcgac
1981 atccggaaga ccaccaagat ggagatcgga ggcgaggagg tggaagtgga agagttcgtg
2041 gaggaggagg acgaggacac agtggcggca ccggtagtag cagtgagtaa gggcgaggaa
2101 gtgatcaaag agttcatgcg gtttaaggtg agaatggaag gaagcatgaa cggccacgag
2161 ttcgaaattg agggagaagg agagggacgg ccctacgagg gcacccagac agccaagctg
2221 aaagtgacaa agggcgggcc tctgccattc gcttgggaca tcctgagccc acagtttatg
2281 tacggctcca aggcctatgt gaaacatcca gctgacattc ccgattataa gaaactgagc
2341 ttccccgagg ggtttaagtg ggaaagagtg atgaacttcg aggacggagg cctggtgact
2401 gtgacccagg acagctccct gcaggatggg accctgatct acaaggtgaa aatgagaggg
2461 acaaattttc cccctgatgg acctgtgatg cagaagaaaa ctatgggatg ggaggcctcc
2521 accgaaaggc tgtatccacg cgacggggtg ctgaaaggag aaatccacca ggctctgaag
2581 ctgaaagatg ggggacatta cctggtggag ttcaagacaa tctacatggc caagaaacct
2641 gtgcagctgc caggctacta ttacgtggac acaaaactgg atatcacttc acacaacgag
2701 gactacacta ttgtggagca gtatgaacgg agcgagggga gacaccatct gttcctgggc
2761 catgggactg gaagtaccgg ctcagggtct agtggaaccg cctcaagcga ggataacaat
2821 atggctgtga tcaaagagtt catgaggttt aaggtgcgca tggagggcag catgaatggg
2881 cacgaatttg agattgaagg agagggcgaa gggaggcctt acgagggcac acagactgcc
2941 aagctgaaag tgaccaaggg aggaccactg cctttcgctt gggatatcct gtctcctcag
3001 tttatgtacg gaagtaaggc ctatgtcaag catcccgctg acattcctga ttacaagaaa
3061 ctgtctttcc cagagggctt taagtgggag agagtgatga attttgaaga tggaggcctg
3121 gtgaccgtga cacaggactc ctctctgcag gatggcactc tgatctacaa agtcaaaatg
3181 cgcggcacca attttccacc cgatgggccc gtgatgcaga agaaaacaat ggggtgggag
3241 gccagcactg aacggctgta tcctagagac ggagtgctga agggcgaaat ccaccaggcc
3301 ctgaagctga aagacggcgg ccactacctg gtggagttca aaaccatcta catggccaag
3361 aaaccagtgc agctgcccgg ctattactat gtggacacca agctggatat cacatcccac
3421 aatgaagact acaccattgt ggaacagtat gagaggtctg aaggacgcca ccatctgttt
3481 ctgtacggca tggatgagct gtataagtaa gaattcgata tcaagcttat cgataatcaa
3541 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
3601 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
3661 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
3721 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
3781 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
3841 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
3901 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
3961 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
4021 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
4081 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc
4141 gggcggccgc tcataaagct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
4201 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
4261 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
4321 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtccg
4381 gactgtacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
4441 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc
4501 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
4561 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
4621 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat
4681 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
4741 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
4801 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
4861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
4921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
4981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
5041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
5101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
5161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
5221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
5281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
5341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
5401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
5461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
5521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
5581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
5641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
5701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
5761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
5821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
5881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
5941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
6001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
6061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
6121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
6181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
6241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
6301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
6361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
6421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
6481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
6541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
6601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
6661 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
6721 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
6781 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
6841 caaagggcga aaaaccgtct atca
//
