LOCUS Exported 5253 bp ds-DNA circular SYN 25-APR-2022 DEFINITION PtdIns(3,4,5)P3 biosensor. ACCESSION . VERSION . KEYWORDS NES-mCherry-PH-BTKx1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5253) TITLE Kinase-independent synthesis of 3-phosphorylated phosphoinositides by a phosphotransferase REFERENCE 2 (bases 1 to 5253) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 25, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5253 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 78..381 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..585 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 536..556 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 612..621 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 669..678 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 675..1382 /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(820..838) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1088..1107 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2058..2179 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2095..2114) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2149..2168 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2186..2641) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2323..2344) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2535..2554 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2668..2772 /gene="bla" /label=AmpR promoter promoter 2774..3131 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2797..2817) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2982..3117 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3044..3063 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3166..3960 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3220..3239) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3830..3849 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4148..4167) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4192..4239 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4568..5156 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5057..5076 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc 61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt 601 cagatccgct agccaccatg gctctgcaga aaaagttgga agagcttgag ctggatgagg 661 caccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt 721 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg 781 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg 841 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg 901 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct 961 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact 1021 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct 1081 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt 1141 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg 1201 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg 1261 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg 1321 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca 1381 agtccggact cagatctcga gctcaagctt cgaattctgc cgcagtgatt ctggagagca 1441 tctttctgaa gcgatcccaa cagaaaaaga aaacatcacc tctaaacttc aagaagcgcc 1501 tgtttctctt gaccgtgcac aaactctcct actatgagta tgactttgaa cgtgggagaa 1561 gaggcagtaa gaagggttca atagatgttg agaagatcac ttgtgttgaa acagtggttc 1621 ctgaaaaaaa tcctcctcca gaaagacaga ttccgagaag aggtgaagag tccagtgaaa 1681 tggagcaaat ttcaatcatt gaaaggttcc cttatccctt ccaggttgta tatgatgaag 1741 ggcctctcta cgtcttctcc ccaactgaag aactaaggaa gcggtggatt caccagctca 1801 aaaacgtaat ccggtacaac agtgatctgg ttcagaaata tcacccttgc ttctggatcg 1861 atgggcagta tctctgctgc tctcagacag ccaaaaatgc tatgggctgc caaattttgg 1921 agaactgagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag 1981 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 2041 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 2101 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 2161 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg 2221 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct 2281 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt 2341 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat 2401 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca 2461 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac 2521 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta 2581 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg 2641 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata 2701 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 2761 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg 2821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 2881 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 2941 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 3001 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 3061 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 3121 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 3181 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 3241 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 3301 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg 3361 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3421 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3481 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 3541 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3601 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 3661 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc 3721 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 3781 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 3841 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 3901 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 3961 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 4021 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 4081 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga 4141 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 4201 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 4261 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 4321 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 4381 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa 4441 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 4501 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 4561 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 4621 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 4681 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 4741 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 4801 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 4861 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 4921 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 4981 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 5041 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 5101 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 5161 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 5221 ttcctgcgtt atcccctgat tctgtggata acc //