LOCUS Exported 6256 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression of iLID fused to mRuby3 and intracellular signaling domain of TrkA (aa 450-799). ACCESSION . VERSION . KEYWORDS iLID-mRuby3-iTrkA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6256) TITLE Construction of light-activated neurotrophin receptors using the improved Light-Induced Dimerizer (iLID) JOURNAL Unpublished REFERENCE 2 (bases 1 to 6256) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6256 /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" regulatory 1121..1130 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1127..1837 /codon_start=1 /product="monomeric red fluorescent protein derived from mRuby2, with improved brightness and photostability (Bajar et al., 2016)" /label=mRuby3 /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV AKYSNLGGGMDELYK" polyA_signal 3044..3165 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3081..3100) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3135..3154 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3172..3627) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3309..3330) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3521..3540 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3654..3758 /gene="bla" /label=AmpR promoter promoter 3760..4117 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3783..3803) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3968..4103 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4030..4049 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4152..4946 /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(4206..4225) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4816..4835 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5134..5153) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5178..5225 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5554..6142 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6043..6062 /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 atctcgagct caagcttcga attcatgggg gagtttctgg caaccacact 661 ggaacggatc gagaaaaatt tcgtgattac tgatccgaga ctgcctgaca acccaatcat 721 ttttgcgagc gattccttcc tgcagctgac agaatattct cgggaagaga tcctggggcg 781 caattgccgt tttctgcagg gacccgagac agaccgtgcc actgttcgga aaatcagaga 841 tgctattgac aaccagactg aagtgaccgt tcagctgatc aattatacca agagcggcaa 901 gaagttctgg aacgtgttcc acctgcagcc gatgcgcgat tataagggcg acgtccagta 961 cttcattggc gtgcagctgg atggcaccga acgtcttcat ggcgccgctg agcgtgaggc 1021 ggtctgcctg atcaaaaaga cagcctttca gattgctgag gcagcgaacg acgaaaatta 1081 ctttggaagc gggagtggga gcccccggga tccaccggtc gccaccatgg tgtctaaggg 1141 cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg 1201 ccaccaattc aaatgcacag gtgaaggaga aggcagaccg tacgagggag tgcaaaccat 1261 gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc 1321 gttcatgtat ggcagccgta cctttatcaa gtacccggcc gacatccctg atttctttaa 1381 acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt 1441 cgtcaccgtc acgcaggaca ccagccttga ggatggcgag ctcgtctaca acgtcaaggt 1501 cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca agggttggga 1561 gcctaataca gagatgatgt atccagcaga tggtggtctg agaggataca ctgacatcgc 1621 actgaaagtt gatggtggtg gccatctgca ctgcaacttc gtgacaactt acaggtcaaa 1681 aaagaccgtc gggaacatca agatgcccgg tgtccatgcc gttgatcacc gcctggaaag 1741 gatcgaggag agtgacaatg aaacctacgt agtgcaaaga gaagtggcag ttgccaaata 1801 cagcaacctt ggtggtggca tggacgagct gtacaagtcc ggactcagat ctggaggctc 1861 cggaggcaaa tttgggatca accgccctgc tgtgctggcg ccagaggatg ggctggccat 1921 gtccctacac ttcatgacac tgggtggcag ttctctttcc cctactgagg gcaaaggctc 1981 cggactccag ggccacatca tggagaaccc acagtacttc agtgatacct gtgtccacca 2041 tatcaagcgc caggacatca ttctcaagtg ggagctaggg gagggagcct ttggaaaggt 2101 ctttcttgct gagtgctaca accttctgaa tgatcaggac aagatgctgg tggctgtcaa 2161 ggcactgaag gagacatctg agaatgctcg tcaggacttc catcgtgagg cagagctgct 2221 caccatgcta cagcaccaac acatcgtacg cttctttgga gtctgcacgg agggtggccc 2281 attgctcatg gtcttcgagt acatgcgcca tggggacctc aaccgtttcc tccggtccca 2341 cggacctgat gcaaaactgc tggctggcgg cgaggatgtg gctcctggtc ctttgggcct 2401 tgggcagctt ctggctgtgg ctagccaggt ggctgctggt atggtgtacc tagccagcct 2461 gcactttgtg caccgggacc tggccacacg caactgtctg gtgggtcagg gactagtggt 2521 caagattgga gacttcggca tgagcaggga catctacagc acagactact accgtgtggg 2581 aggtcggacc atgctgccca tccgctggat gcctccagag agcatcctct accgcaagtt 2641 cagcaccgag agtgatgtgt ggagcttcgg ggtggtgctc tgggagatct tcacctatgg 2701 aaagcaaccc tggtaccagc tctccaacac tgaggcgatc gagtgcatca cgcagggccg 2761 ggagctggag cggccgcgcg cctgccctcc tgatgtctac gccatcatgc gcggctgctg 2821 gcagcgtgag ccgcaacagc gcctcagcat gaaggatgtg cacgcgcggc tgcaggcctt 2881 ggcacaggcg ccaccgagtt acctggacgt tctgggttag aagcggccgc gactctagat 2941 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 3001 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 3061 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 3121 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa 3181 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 3241 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 3301 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 3361 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 3421 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 3481 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 3541 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 3601 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa 3661 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 3721 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct 3781 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3841 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 3901 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 3961 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 4021 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 4081 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag 4141 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 4201 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 4261 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 4321 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4381 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4441 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 4501 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4561 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4621 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga 4681 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4741 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4801 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4861 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4921 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 4981 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 5041 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 5101 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga 5161 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt 5221 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc 5281 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg 5341 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta 5401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 5461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 5521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 5581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 5641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 5701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 5761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 5821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 5881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 5941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 6001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 6061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 6121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 6181 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 6241 tattaccgcc atgcat //