LOCUS Exported 7750 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent indicator for PI(3,4,5)P3. ACCESSION . VERSION . KEYWORDS pcDNA3.1_AktPH-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7750) AUTHORS Kawano F, Suzuki H, Furuya A, Sato M TITLE Engineered pairs of distinct photoswitches for optogenetic control of cellular proteins. JOURNAL Nat Commun. 2015 Feb 24;6:6256. doi: 10.1038/ncomms7256. PUBMED 25708714 REFERENCE 2 (bases 1 to 7750) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7750 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1475..2185 /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(1620..1638) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1888..1907 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(2246..2266) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2664..2675) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2872..2891) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2937..2992 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2991..3010) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3344..3361) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3350..3574 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3620..4048 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3707..3726) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3917..3938 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4057..4077) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4062..4391 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4242..4377 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4304..4323 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4458..5252 /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(4512..4531) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5122..5141 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5426..5547 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5463..5482) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5517..5536 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5596..5612) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5596..5612) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5609..5631) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5620..5636 /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(5644..5674) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5689..5710 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5827..5844) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5998..6583) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6078..6097) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6754..7614) /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 7377..7396 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7615..7719) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttgccg ccaccatgag cgacgtggct attgtgaagg agggttggct 961 gcacaaacga ggggagtaca tcaagacctg gcggccacgc tacttcctcc tcaagaatga 1021 tggcaccttc attggctaca aggagcggcc gcaggatgtg gaccaacgtg aggctcccct 1081 caacaacttc tctgtggcgc agtgccagct gatgaagacg gagcggcccc ggcccaacac 1141 cttcatcatc cgctgcctgc agtggaccac tgtcatcgaa cgcaccttcc atgtggagac 1201 tcctgaggag cgggaggagt ggacaaccgc catccagact gtggctgacg gcctcaagaa 1261 gcaggaggag gaggagatgg acttccggtc gggctcaccc agtgacaact caggggctga 1321 agagatggag gtgtccctgg ccaagcccaa gcaccgcgtg accatgaacg agtttgagta 1381 cctgaagctg ctgggcaagg gcactttcgg caaggtggaa ttcggagggt ctggaggggg 1441 tggctcaggt ggcggcggaa gtggcggaag atctatggtg agcaagggcg aggaggataa 1501 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg 1561 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc 1621 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca 1681 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa 1741 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt 1801 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct 1861 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga 1921 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag 1981 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa 2041 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca 2101 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg 2161 cggcatggac gagctgtaca agtaatctag atgatcaacc tctggattac aaaatttgtg 2221 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 2281 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 2341 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 2401 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 2461 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 2521 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 2581 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg 2641 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 2701 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 2761 ccctttgggc cgcctccccg catcggtaca attcactcct caggtgcagg ctgcctatca 2821 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc 2881 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata 2941 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag 3001 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc 3061 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta 3121 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta 3181 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt 3241 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc 3301 tcttctctta tggagatcgg gcccgtttaa acccgctgat cagcctcgac tgtgccttct 3361 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc 3421 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt 3481 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat 3541 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg 3601 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 3661 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 3721 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 3781 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 3841 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 3901 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 3961 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 4021 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 4081 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 4141 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4201 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 4261 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4321 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4381 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag 4441 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 4501 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 4561 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 4621 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 4681 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 4741 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 4801 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 4861 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 4921 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 4981 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 5041 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 5101 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 5161 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 5221 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga 5281 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag 5341 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct 5401 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata 5461 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5521 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag 5581 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 5641 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 5701 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 5761 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 5821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 5881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 5941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 6001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 6061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 6121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 6181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 6241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 6301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 6361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 6421 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt 6541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 6601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 6661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 6721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 6781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 6841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 6901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 6961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 7381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 7441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 7501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 7561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 7621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 7681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 7741 acctgacgtc //