LOCUS Exported 6861 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCry2PHR-mCh-LRP6c SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6861) AUTHORS Bugaj LJ, Choksi AT, Mesuda CK, Kane RS, Schaffer DV TITLE Optogenetic protein clustering and signaling activation in mammalian cells. JOURNAL Nat Methods. 2013 Feb 3. doi: 10.1038/nmeth.2360. PUBMED 23377377 REFERENCE 2 (bases 1 to 6861) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6861 /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..2847 /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 EGRHSTGGMDELY" primer_bind complement(2288..2306) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2556..2575 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 3649..3770 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3686..3705) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3740..3759 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3777..4232) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3914..3935) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4126..4145 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4259..4363 /gene="bla" /label=AmpR promoter promoter 4365..4722 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4388..4408) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4573..4708 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4635..4654 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4757..5551 /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(4811..4830) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5421..5440 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5739..5758) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5783..5830 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6159..6747 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6648..6667 /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 gctgtacgat ggtggaggag gtagtggagg aggaggatca 2881 aggatgttgt gtccacgtat gaagggagat ggggaaacta tgactaatga ctatgtagtt 2941 catggaccag cttctgtgcc tcttggttat gtgccacacc caagttcttt gtcaggatct 3001 cttccaggaa tgtctcgagg taaatcaatg atcagctccc tcagtatcat ggggggaagc 3061 agtggacccc cctatgaccg agcccatgtt acaggagcat catcaagtag ttcttcaagc 3121 accaaaggca cttacttccc tgcaattttg aaccctccac catccccagc cacagagcga 3181 tcacattaca ctatggaatt tggatattct tcaaacagtc cttccactca taggtcatac 3241 agctacaggc catatagcta ccggcacttt gcacccccca ccacaccctg cagcacagat 3301 gtttgtgaca gtgactatgc tcctagtcgg agaatgacct cagtggcaac agccaagggc 3361 tataccagtg acttgaacta tgattcagaa cctgtgcccc cacctcccac accccgaagc 3421 caatacttgt cagcagagga gaactatgaa agctgcccac cttctccata cacagagagg 3481 agctattctc atcacctcta cccaccgcca ccctctccct gtacagactc ctcctgactc 3541 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca 3601 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 3661 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 3721 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat 3781 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 3841 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 3901 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 3961 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc 4021 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 4081 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 4141 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 4201 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg 4261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 4321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac 4381 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 4441 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 4501 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 4561 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 4621 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 4681 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg 4741 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 4801 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 4861 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 4921 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt 4981 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 5041 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 5101 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 5161 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 5221 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 5281 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 5341 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 5401 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 5461 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 5521 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 5581 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 5641 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 5701 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata 5761 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg 5821 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 5881 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 5941 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca 6001 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 6061 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 6121 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 6181 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 6241 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 6301 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 6361 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 6421 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 6481 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 6541 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 6601 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 6661 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 6721 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 6781 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 6841 aaccgtatta ccgccatgca t //