LOCUS Exported 7301 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression vector for ChrimsonR-tdTomato. ACCESSION . VERSION . KEYWORDS pCAG-ChrimsonR-tdT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7301) 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 7301) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7301 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 385..660 /label=chicken beta-actin promoter intron 661..1674 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1598..1620 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1682..1701 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1735..1744 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1741..2790 /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" primer_bind complement(4323..4342) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4388..4443 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4442..4461) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4804..4820) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4804..4820) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4817..4839) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4828..4844 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4852..4882) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4897..4918 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4977..5172 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5023..5158 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5085..5104 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5178..5312 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5215..5234) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5269..5288 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5380..5397) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5551..6139) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5631..5650) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6310..7170) /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 6933..6952 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7171..7275) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca 421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag 481 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg 541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt 601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg 661 ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg ccgcccgccc 721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg 781 ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct gcgtgaaagc 841 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg 901 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc 961 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc 1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc 1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag 1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc 1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc 1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg 1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctg agccgccacc 1741 atggctgagc tgatcagcag cgccaccaga tctctgtttg ccgccggagg catcaaccct 1801 tggcctaacc cctaccacca cgaggacatg ggctgtggag gaatgacacc tacaggcgag 1861 tgcttcagca ccgagtggtg gtgtgaccct tcttacggac tgagcgacgc cggatacgga 1921 tattgcttcg tggaggccac aggcggctac ctggtcgtgg gagtggagaa gaagcaggct 1981 tggctgcaca gcagaggcac accaggagaa aagatcggcg cccaggtctg ccagtggatt 2041 gctttcagca tcgccatcgc cctgctgaca ttctacggct tcagcgcctg gaaggccact 2101 tgcggttggg aggaggtcta cgtctgttgc gtcgaggtgc tgttcgtgac cctggagatc 2161 ttcaaggagt tcagcagccc cgccacagtg tacctgtcta ccggcaacca cgcctattgc 2221 ctgcgctact tcgagtggct gctgtcttgc cccgtgatcc tgatcagact gagcaacctg 2281 agcggcctga agaacgacta cagcaagcgg accatgggcc tgatcgtgtc ttgcgtggga 2341 atgatcgtgt tcggcatggc cgcaggactg gctaccgatt ggctcaagtg gctgctgtat 2401 atcgtgtctt gcatctacgg cggctacatg tacttccagg ccgccaagtg ctacgtggaa 2461 gccaaccaca gcgtgcctaa aggccattgc cgcatggtcg tgaagctgat ggcctacgct 2521 tacttcgcct cttggggcag ctacccaatc ctctgggcag tgggaccaga aggactgctg 2581 aagctgagcc cttacgccaa cagcatcggc cacagcatct gcgacatcat cgccaaggag 2641 ttttggacct tcctggccca ccacctgagg atcaagatcc acgagcacat cctgatccac 2701 ggcgacatcc ggaagaccac caagatggag atcggaggcg aggaggtgga agtggaagag 2761 ttcgtggagg aggaggacga ggacacagtg gcggcaccgg tagtagcagt gagtaagggc 2821 gaggaagtga tcaaagagtt catgcggttt aaggtgagaa tggaaggaag catgaacggc 2881 cacgagttcg aaattgaggg agaaggagag ggacggccct acgagggcac ccagacagcc 2941 aagctgaaag tgacaaaggg cgggcctctg ccattcgctt gggacatcct gagcccacag 3001 tttatgtacg gctccaaggc ctatgtgaaa catccagctg acattcccga ttataagaaa 3061 ctgagcttcc ccgaggggtt taagtgggaa agagtgatga acttcgagga cggaggcctg 3121 gtgactgtga cccaggacag ctccctgcag gatgggaccc tgatctacaa ggtgaaaatg 3181 agagggacaa attttccccc tgatggacct gtgatgcaga agaaaactat gggatgggag 3241 gcctccaccg aaaggctgta tccacgcgac ggggtgctga aaggagaaat ccaccaggct 3301 ctgaagctga aagatggggg acattacctg gtggagttca agacaatcta catggccaag 3361 aaacctgtgc agctgccagg ctactattac gtggacacaa aactggatat cacttcacac 3421 aacgaggact acactattgt ggagcagtat gaacggagcg aggggagaca ccatctgttc 3481 ctgggccatg ggactggaag taccggctca gggtctagtg gaaccgcctc aagcgaggat 3541 aacaatatgg ctgtgatcaa agagttcatg aggtttaagg tgcgcatgga gggcagcatg 3601 aatgggcacg aatttgagat tgaaggagag ggcgaaggga ggccttacga gggcacacag 3661 actgccaagc tgaaagtgac caagggagga ccactgcctt tcgcttggga tatcctgtct 3721 cctcagttta tgtacggaag taaggcctat gtcaagcatc ccgctgacat tcctgattac 3781 aagaaactgt ctttcccaga gggctttaag tgggagagag tgatgaattt tgaagatgga 3841 ggcctggtga ccgtgacaca ggactcctct ctgcaggatg gcactctgat ctacaaagtc 3901 aaaatgcgcg gcaccaattt tccacccgat gggcccgtga tgcagaagaa aacaatgggg 3961 tgggaggcca gcactgaacg gctgtatcct agagacggag tgctgaaggg cgaaatccac 4021 caggccctga agctgaaaga cggcggccac tacctggtgg agttcaaaac catctacatg 4081 gccaagaaac cagtgcagct gcccggctat tactatgtgg acaccaagct ggatatcaca 4141 tcccacaatg aagactacac cattgtggaa cagtatgaga ggtctgaagg acgccaccat 4201 ctgtttctgt acggcatgga tgagctgtat aagtaaagcg gccgcactcc tcaggtgcag 4261 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga 4321 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact 4381 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc 4441 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt 4501 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga 4561 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg 4621 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta 4681 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc 4741 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc 4801 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4861 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 4921 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat 4981 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 5041 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 5101 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 5161 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5221 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5281 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc 5341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 5401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 5461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 5521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 5581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 5641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 5701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 5761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 5821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 6001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 6061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 6121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 6181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 6241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 6301 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 6361 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 6421 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 6481 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 6541 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 6601 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 6661 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 6721 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 6781 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 6841 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 6901 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 6961 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 7021 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 7081 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 7141 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 7201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 7261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct g //