LOCUS Exported 8364 bp ds-DNA circular SYN 15-APR-2022 DEFINITION Soluble light-dependent effector protein (nucleolin) for optodroplet formation or recruitment to CIBN localizers. ACCESSION . VERSION . KEYWORDS pPHR-mCherry-hNCL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8364) TITLE Rippe Lab Plasmids 2022 REFERENCE 2 (bases 1 to 8364) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8364 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 625..2118 /codon_start=1 /product="N-terminal photolyase homology region of Arabidopsis thaliana cryptochrome 2" /label=CRY2PHR /note="binds to CIB1 upon exposure to blue light (Kennedy et al., 2010)" /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA" regulatory 2137..2146 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2143..2850 /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(2288..2306) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2556..2575 /label=mCherry-F /note="mCherry, forward primer" protein_bind 3460..3471 /label=ZF binding site /bound_moiety="CCR5TC zinc-finger domain" /note="target sequence for the CCR5TC zinc-finger domain from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 2008; Gross et al., 2013)" protein_bind 3478..3489 /label=ZF binding site /bound_moiety="CCR5TC zinc-finger domain" /note="target sequence for the CCR5TC zinc-finger domain from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 2008; Gross et al., 2013)" CDS 4537..4548 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 5152..5273 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5189..5208) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5243..5262 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5280..5735) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5417..5438) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5629..5648 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5762..5866 /gene="bla" /label=AmpR promoter promoter 5868..6225 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5891..5911) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6076..6211 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6138..6157 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6260..7054 /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(6314..6333) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6924..6943 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7242..7261) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7286..7333 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7662..8250 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8151..8170 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgaggc caccatgaag atggacaaaa agactatagt ttggtttaga 661 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt 721 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca 781 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct 841 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt 901 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac 961 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat 1021 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat 1081 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg 1141 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta 1201 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca 1261 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat 1321 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat 1381 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg 1441 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt 1501 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg 1561 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga 1621 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga 1681 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt 1741 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt 1801 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc 1861 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa 1921 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct 1981 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta 2041 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag 2101 atcatgatcg gagcagcagc ccgggatcca ccggtcgcca ccatggtgag caagggcgag 2161 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc 2221 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc 2281 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg 2341 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac 2401 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac 2461 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag 2521 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg 2581 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc 2641 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac 2701 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc 2761 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac 2821 tccaccggcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg 2881 aattctgcag tcgacatggt gaagctcgcg aaggcaggta aaaatcaagg tgaccccaag 2941 aaaatggctc ctcctccaaa ggaggtagaa gaagatagtg aagatgagga aatgtcagaa 3001 gatgaagaag atgatagcag tggagaagag gtcgtcatac ctcagaagaa aggcaagaag 3061 gctgctgcaa cctcagcaaa gaaggtggtc gtttccccaa caaaaaaggt tgcagttgcc 3121 acaccagcca agaaagcagc tgtcactcca ggcaaaaagg cagcagcaac acctgccaag 3181 aagacagtta caccagccaa agcagttacc acacctggca agaagggagc cacaccaggc 3241 aaagcattgg tagcaactcc tggtaagaag ggtgctgcca tcccagccag gggggcaaag 3301 aatggcaaga atgccgagga ggaagacagt gatgaagagg aggatgatga cagtgaggag 3361 gatgaggagg atgacgagga cgaggatgag gatgaagatg aaattgaacc agcagcgatg 3421 aaagcagcag ctgctgcccc tgcctcagag gatgaggacg atgaggatga cgaagatgat 3481 gaggatgacg atgacgatga ggaagatgac tctgaagaag aagctatgga gactacacca 3541 gccaaaggaa agaaagctgc aaaagttgtt cctgtgaaag ccaagaacgt ggctgaggat 3601 gaagatgaag aagaggatga tgaggacgag gatgacgacg acgacgaaga tgatgaagat 3661 gatgatgatg aagatgatga ggaggaggaa gaagaggagg aggaagagcc tgtcaaagaa 3721 gcacctggaa aacgaaagaa ggaaatggcc aaacagaaag cagctcctga agccaagaaa 3781 cagaaagtgg aaggcacaga accgactacg gctttcaatc tctttgttgg aaacctaaac 3841 tttaacaaat ctgctcctga attaaaaact ggtatcagcg atgtttttgc taaaaatgat 3901 cttgctgttg tggatgtcag aattggtatg actaggaaat ttggttatgt ggattttgaa 3961 tctgctgaag acctggagaa agcgttggaa ctcactggtt tgaaagtctt tggcaatgaa 4021 attaaactag agaaaccaaa aggaaaagac agtaagaaag agcgagatgc gagaacactt 4081 ttggctaaaa atctccctta caaagtcact caggatgaat tgaaagaagt gtttgaagat 4141 gctgcggaga tcagattagt cagcaaggat gggaaaagta aagggattgc ttatattgaa 4201 tttaagacag aagctgatgc agagaaaacc tttgaagaaa agcagggaac agagatcgat 4261 gggcgatcta tttccctgta ctatactgga gagaaaggtc aaaatcaaga ctatagaggt 4321 ggaaagaata gcacttggag tggtgaatca aaaactctgg ttttaagcaa cctctcctac 4381 agtgcaacag aagaaactct tcaggaagta tttgagaaag caacttttat caaagtaccc 4441 cagaaccaaa atggcaaatc taaagggtat gcatttatag agtttgcttc attcgaagac 4501 gctaaagaag ctttaaattc ctgtaataaa agggaaattg agggcagagc aatcaggctg 4561 gagttgcaag gacccagggg atcacctaat gccagaagcc agccatccaa aactctgttt 4621 gtcaaaggcc tgtctgagga taccactgaa gagacattaa aggagtcatt tgacggctcc 4681 gttcgggcta ggatagttac tgaccgggaa actgggtcct ccaaagggtt tggttttgta 4741 gacttcaaca gtgaggagga tgccaaagct gccaaggagg ccatggaaga cggtgaaatt 4801 gatggaaata aagttacctt ggactgggcc aaacctaagg gtgaaggtgg cttcgggggt 4861 cgtggtggag gcagaggcgg ctttggagga cgaggtggtg gtagaggagg ccgaggagga 4921 tttggtggca gaagccgggg aggctttgga gggcgaggag gcttccgagg aggcagagga 4981 ggaggaggtg accacaagcc acaaggaaag aagacgaagt ttgaataggg gatccaccgg 5041 atctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 5101 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 5161 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 5221 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta 5281 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 5341 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 5401 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 5461 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta 5521 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 5581 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 5641 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 5701 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt 5761 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 5821 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag 5881 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 5941 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 6001 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 6061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 6121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc 6181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac 6241 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 6301 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 6361 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 6421 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg 6481 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 6541 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 6601 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 6661 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 6721 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 6781 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 6841 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 6901 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 6961 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 7021 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 7081 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 7141 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 7201 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca 7261 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt 7321 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca 7381 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca 7441 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc 7501 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 7561 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 7621 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 7681 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 7741 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 7801 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 7861 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 7921 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 7981 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 8041 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 8101 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 8161 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 8221 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 8281 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 8341 gataaccgta ttaccgccat gcat //