LOCUS Exported 5423 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION EGFP-mCherry Met71GAG reporter for methionine misincorporation. ACCESSION . VERSION . KEYWORDS GAG-Pan SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5423) AUTHORS Gomes AC, Kordala AJ, Strack R, Wang X, Geslain R, Delaney K, Clark WC, Keenan R, Pan T TITLE A dual fluorescent reporter for the investigation of methionine mistranslation in live cells. JOURNAL RNA. 2016 Mar;22(3):467-76. doi: 10.1261/rna.054163.115. Epub 2016 Jan 4. PUBMED 26729921 REFERENCE 2 (bases 1 to 5423) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5423 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 120..423 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 424..627 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 578..598 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 694..703 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 700..1416 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(745..766) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1006..1025) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1353..1374 /label=EGFP-C /note="EGFP, forward primer" regulatory 1432..1441 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1438..2148 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFEYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1583..1601) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1851..1870 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2270..2391 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2307..2326) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2361..2380 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2398..2853) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2535..2556) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2747..2766 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2880..2984 /gene="bla" /label=AmpR promoter promoter 2986..3343 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3009..3029) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3194..3329 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3256..3275 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3378..4172 /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(3432..3451) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4042..4061 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4360..4379) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4404..4451 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4780..5368 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5269..5288 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt 61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 601 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac 661 cggactcaga tctcgagctc aagcttcgaa ttcgccacca tggtgagcaa gggcgaggag 721 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag 781 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc 841 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac 901 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc 961 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac 1021 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag 1081 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac 1141 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag 1201 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc 1261 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag 1321 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc 1381 gccgggatca ctctcggcat ggacgagctg tacaagtcgg atccactagt cgccaccatg 1441 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg 1501 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc 1561 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc 1621 gcctgggaca tcctgtcccc tcagttcgag tacggctcca aggcctacgt gaagcacccc 1681 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg 1741 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc 1801 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg 1861 cagaagaaga ctatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc 1921 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag 1981 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac 2041 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc 2101 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagc ggccgcgact 2161 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc 2221 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat 2281 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 2341 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa 2401 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 2461 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 2521 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 2581 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat 2641 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 2701 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 2761 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac 2821 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc 2881 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 2941 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa 3001 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 3061 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 3121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 3181 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 3301 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag 3361 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 3421 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 3481 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 3541 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg 3601 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 3661 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 3721 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 3781 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 3841 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 3901 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 3961 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 4021 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 4081 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 4141 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga 4201 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag 4261 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct 4321 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat 4381 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg 4441 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc 4501 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag 4561 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc 4621 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 4681 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 4741 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 4801 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 4861 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 4921 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 4981 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 5041 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 5101 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 5161 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 5221 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 5281 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 5341 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 5401 ggccttttgc tggccttttg ctc //