LOCUS Exported 8152 bp ds-DNA circular SYN 25-JUL-2022 DEFINITION Optically controlled eDrEGFR expression in mammalian cells. ACCESSION . VERSION . KEYWORDS eDrEGFR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8152) TITLE HOLD - Verkhusha lab plasmids 2022 REFERENCE 2 (bases 1 to 8152) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 25, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8152 /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" sig_peptide 597..659 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader CDS 660..1361 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG RHSTGGMDELYK" primer_bind complement(799..817) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1067..1086 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 4940..5061 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4977..4996) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5031..5050 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5068..5523) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5205..5226) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5417..5436 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5550..5654 /gene="bla" /label=AmpR promoter promoter 5656..6013 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5679..5699) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5864..5999 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5926..5945 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6048..6842 /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(6102..6121) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6712..6731 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7030..7049) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7074..7121 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7450..8038 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7939..7958 /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 gctagcatgg 601 agacagacac actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca 661 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca 721 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct 781 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct 841 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg 901 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga 961 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt 1021 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga 1081 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga 1141 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca 1201 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca 1261 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg 1321 agggccgcca ctccaccggc ggcatggacg agctgtacaa ggaattcagt gctggtagtg 1381 ctggtagtgc tggcaccggt atgagccggg acccgttgcc cttttttcca ccgctttacc 1441 ttggtggccc ggaaattacc accgagaact gcgagcgcga gccgattcat attcccggca 1501 gcatccagcc gcacggcgcc ctgctcactg ccgacgggca cagcggcgag gtgctccaga 1561 tgagcctcaa cgcggccact tttctgggac aggaacccac agtgctgcgc ggacagaccc 1621 tcgccgcact gctgcccgag cagtggcccg cgctgcaagc ggccctgccc cccggctgcc 1681 ccgacgccct gcaataccgc gcaacgctgg actggcctgc cgccgggcac ctttcgctga 1741 cggtgcaccg ggtcggcgag ttgctgattc tggagttcga gccgacggag gcctgggaca 1801 gcaccgggcc gcacgcgctg cgcaacgcga tgttcgcgct cgaaagtgcc cccaacctgc 1861 gggcgctggc cgaggtggcg acccagacgg tccgcgagct gacgggcttt gaccgggtga 1921 tgctctacaa atttgccccc gacgccaccg gcgaagtgat tgccgaggcc cgccgtgagg 1981 ggctgcacgc ctttctgggc caccgttttc ccgcgtcgga cattccggcg caggcccgcg 2041 cgctctacac ccggcacctg ctgcgcctga ccgccgacac ccgcgccgcc gccgtgccgc 2101 tcgatcccgt cctcaacccg cagacgaatg cgcccacccc gctgggcggc gccgtgctgc 2161 gcgccacctc gcccatgcac atgcagtacc tgcggaacat gggcgtcggg tcgagcctgt 2221 cggtgtcggt ggtggtcggc ggccagctct ggggcctgat cgcctgccac caccagacgc 2281 cctacgtgtt gccgcccgac ctgcgaacca cgctcgaata cctgggccgc ttgctgagcc 2341 tgcaagttca ggtcaaggaa gcggcggacg tggcggcctt tcgccagagc ctgcgggagc 2401 accacgcgcg ggtggccctc gcggcggcgc actcgctctc gccgcacgac accctcagtg 2461 acccggcgct tgacctgctg ggcctgatgc gggccggggg cctgattctg cgtttcgagg 2521 gccgctggca gacgttgggt gaagtgccgc ctgccccggc ggtggacgcg ctgctggcgt 2581 ggctcgaaac ccagccgggc gccctggtcc agaccgacgc gctaggccaa ctgtggcccg 2641 ccggcgccga tctcgccccc agcgcagcgg gcctgctcgc catcagcgtg ggcgagggct 2701 ggtcggagtg cctcgtctgg ctgcggcccg aactgcggct ggaggtcgcc tggggcgggg 2761 ccactcctga ccaggcgaaa gacgacctcg ggccgcgcca ctcattcgac acctacctcg 2821 aagaaaaacg cggctacgcc gagccctggc atcccggcga aatcgaggag gcgcaggatc 2881 tacgtgacac attgaccggg gcgctgggcg agcagctcga gatcgccact gggatggtgg 2941 gggccctcct cttgctgctg gtggtggccc tggggatcgg cctcttcatg cgaaggcgcc 3001 acatcgttcg gaagcgcacg ctgcggaggc tgctgcagga gagggagctt gtggagcctc 3061 ttacacccag tggagaagct cccaaccaag ctctcttgag gatcttgaag gaaactgaat 3121 tcaaaaagat caaagtgctg ggctccggtg cgttcggcac ggtgtataag ggactctgga 3181 tcccagaagg tgagaaagtt aaaattcccg tcgctatcaa ggaattaaga gaagcaacat 3241 ctccgaaagc caacaaggaa atcctcgatg aagcctacgt gatggccagc gtggacaacc 3301 cccacgtgtg ccgcctgctg ggcatctgcc tcacctccac cgtgcaactc atcacgcagc 3361 tcatgccctt cggctgcctc ctggactatg tccgggaaca caaagacaat attggctccc 3421 agtacctgct caactggtgt gtgcagatcg caaagggcat gaactacttg gaggaccgtc 3481 gcttggtgca ccgcgacctg gcagccagga acgtactggt gaaaacaccg cagcatgtca 3541 agatcacaga ttttgggctg gccaaactgc tgggtgcgga agagaaagaa taccatgcag 3601 aaggaggcaa agtgcctatc aagtggatgg cattggaatc aattttacac agaatctata 3661 cccaccagag tgatgtctgg agctacgggg tgaccgtttg ggagttgatg acctttggat 3721 ccaagccata tgacggaatc cctgccagcg agatctcctc catcctggag aaaggagaac 3781 gcctccctca gccacccata tgtaccatcg atgtctacat gatcatggtc aagtgctgga 3841 tgatagacgc agatagtcgc ccaaagttcc gtgagttgat catcgaattc tccaaaatgg 3901 cccgagaccc ccagcgctac cttgtcattc agggggatga aagaatgcat ttgccaagtc 3961 ctacagactc caacttctac cgtgccctga tggatgaaga agacatggac gacgtggtgg 4021 atgccgacga gtacctcatc ccacagcagg gcttcttcag cagcccctcc acgtcacgga 4081 ctcccctcct gagctctctg agtgcaacca gcaacaattc caccgtggct tgcattgata 4141 gaaatgggct gcaaagctgt cccatcaagg aagacagctt cttgcagcga tacagctcag 4201 accccacagg cgccttgact gaggacagca tagacgacac cttcctccca gtgcctgaat 4261 acataaacca gtccgttccc aaaaggcccg ctggctctgt gcagaatcct gtctatcaca 4321 atcagcctct gaaccccgcg cccagcagag acccacacta ccaggacccc cacagcactg 4381 cagtgggcaa ccccgagtat ctcaacactg tccagcccac ctgtgtcaac agcacattcg 4441 acagccctgc ccactgggcc cagaaaggca gccaccaaat tagcctggac aaccctgact 4501 accagcagga cttctttccc aaggaagcca agccaaatgg catctttaag ggctccacag 4561 ctgaaaatgc agaataccta agggtcgcgc cacaaagcag tgaatttatt ggagcatcta 4621 gaggcggtgg tggcagcgga ggagggggga gcggcggcgg tggcagtggc ggcggcggta 4681 gtcgcagccg ctttgtgaaa aaagatagcg ccggcagtgc gggtagtgcg gggagcgccg 4741 gtagctatct ggcgaacgaa attctgtggg gcagcgctgg tagcgcagga agcgccggca 4801 gtgcagggtt ttgctatgaa aacgaagtgg cgctgtcata atcagccata ccacatttgt 4861 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat 4921 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa 4981 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 5041 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg 5101 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 5161 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga 5221 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 5281 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 5341 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 5401 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 5461 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 5521 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 5581 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 5641 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg 5701 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 5761 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 5821 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 5881 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 5941 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 6001 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag 6061 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 6121 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 6181 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag 6241 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 6301 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 6361 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 6421 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 6481 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 6541 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg 6601 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 6661 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 6721 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 6781 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 6841 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 6901 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 6961 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg 7021 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat 7081 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc 7141 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt 7201 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa 7261 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt 7321 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 7381 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 7441 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 7501 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 7561 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 7621 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 7681 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 7741 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 7801 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 7861 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 7921 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 7981 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 8041 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 8101 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at //