LOCUS Exported 7186 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Blue opsin tagged with mcherry on the C terminus. ACCESSION . VERSION . KEYWORDS Blue opsin mcherry ---- pcDNA3.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7186) AUTHORS Karunarathne WK, Giri L, Kalyanaraman V, Gautam N TITLE Optically triggering spatiotemporally confined GPCR activity in a cell and programming neurite initiation and extension. JOURNAL Proc Natl Acad Sci U S A. 2013 Apr 23;110(17):E1565-74. doi: 10.1073/pnas.1220697110. Epub 2013 Mar 11. PUBMED 23479634 REFERENCE 2 (bases 1 to 7186) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7186 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS complement(1707..1724) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS complement(1946..1957) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" regulatory 2031..2040 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2037..2747 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(2182..2200) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2450..2469 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(2779..2796) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2785..3009 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3055..3483 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3142..3161) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3352..3373 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3492..3512) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3497..3826 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3677..3812 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3739..3758 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3893..4687 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3947..3966) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4557..4576 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4861..4982 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4898..4917) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4952..4971 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5031..5047) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5031..5047) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5044..5066) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5055..5071 /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(5079..5109) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5124..5145 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5262..5279) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5433..6021) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5513..5532) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6192..7052) /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 6815..6834 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7053..7157) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta 901 gcgtttaaac ttaagcttgg taccgagctc ggatccacta gtccagtgtg gtggaattca 961 tgcgcaagat gtccgaggag gagttctacc tgttcaagaa catctcgagc gtcggccctt 1021 gggacgggcc ccagtaccac atcgctcctg tctgggcttt ctacctccag gctgctttca 1081 tggggaccgt gttcctcatc ggtttccctc tgaatgccat ggtgctcgtg gccaccctgc 1141 gttacaagaa gctgcgacag cctttaaact acatcctggt gaacgtgtcc ttcggtggat 1201 tcctgctgtg tatcttctcc gtgttccctg tgttcgtggc tagctgcaac ggttacttcg 1261 tgttcggacg tcacgtgtgt gctctggagg gtttcctagg taccgtggct ggtctggtca 1321 ccgggtggtc tctggccttc ctggccttcg aacgctacat cgtcatatgc aagcctttcg 1381 gtaacttcag gttctcctcc aagcacgcgt taaccgtagt actggctacc tggacaatcg 1441 gtatcggtgt atcgatccct cccttcttcg gttggtctcg cttcattccg gaaggccttc 1501 agtgcagctg tggccctgac tggtacaccg tgggtacaaa gtaccgatcg gagagctaca 1561 cctggttcct gttcatcttc tgcttcatcg tgcctctgag cctgatctgc ttctcctaca 1621 cccaactgct gcgagctctg aaggctgtgg ctgctcaaca gcaggagagc gctaccaccc 1681 agaaggctga aagggaggtg tctagaatgg tggtggtgat ggtgggatcc ttctgcgtgt 1741 gctacgtacc ctacgctgct ttcgccatgt acatggtgaa caacaggaac cacggtttag 1801 atctgcgact ggtgacaatc ccttcgttct tctccaagtc cgcatgcatt tacaacccta 1861 tcatctactg cttcatgaac aagcagttcc aagcttgcat catgaagatg gtgtgcggta 1921 aggctatgac cgacgagagc gacacctgca gctcccagaa gaccgaggtg tcgaccgtgt 1981 cctccaccca ggttggccct aacgagacct cccaggtggc tcctgctgcg gccgccatgg 2041 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc 2101 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 2161 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg 2221 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg 2281 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga 2341 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg 2401 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc 2461 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc 2521 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg 2581 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca 2641 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg 2701 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaatct agagggcccg 2761 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2821 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2881 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2941 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 3001 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 3061 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3121 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3181 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 3241 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3301 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 3361 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 3421 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 3481 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3601 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 3781 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 3841 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 3901 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 3961 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 4021 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 4081 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 4141 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 4201 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 4261 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 4321 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 4381 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 4441 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 4501 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 4561 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 4621 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4681 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 4741 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4801 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 4861 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4921 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4981 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 5041 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 5101 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 5161 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 5221 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 5281 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 5341 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 5401 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 5461 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 5521 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5581 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 5641 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5701 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5761 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 5821 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 5881 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 5941 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 6001 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 6061 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 6121 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 6181 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 6241 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 6301 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 6361 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 6421 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 6481 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 6541 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 6601 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 6661 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 6721 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 6781 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 6841 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 6901 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 6961 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 7021 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 7081 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 7141 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacg //