LOCUS Exported 4729 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: Actin, Excitation: 488, Emission: 507. ACCESSION . VERSION . KEYWORDS mEGFP-Lifeact-7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4729) TITLE Michael Davidson Empty Backbones REFERENCE 2 (bases 1 to 4729) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4729 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 39..342 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 343..546 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 497..517 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 575..584 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 581..631 /codon_start=1 /product="peptide that binds filamentous actin (Riedl et al., 2008)" /label=Lifeact /note="first 17 amino acids of budding yeast Abp140" /translation="MGVADLIKKFESISKEE" regulatory 647..656 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 653..1372 /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(698..719) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(959..978) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1306..1327 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1495..1616 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1532..1551) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1586..1605 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1623..2078) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1760..1781) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1972..1991 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2105..2209 /gene="bla" /label=AmpR promoter promoter 2211..2568 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2234..2254) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2419..2554 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2481..2500 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2603..3397 /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(2657..2676) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3267..3286 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3585..3604) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3629..3676 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4005..4593 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4494..4513 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 61 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 121 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 181 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc 241 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct 301 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag 361 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt 421 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac 481 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc 541 agagctggtt tagtgaaccg tcagatccgc tagcgccacc atgggcgtgg ccgacttgat 601 caagaagttc gagtccatct ccaaggagga gggggatcca ccggtcgcca ccatggtgag 661 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 721 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 781 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 841 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 901 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 961 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 1021 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 1081 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 1141 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 1201 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 1261 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 1321 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg 1381 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 1441 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 1501 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 1561 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc 1621 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 1681 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 1741 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 1801 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 1861 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 1921 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 1981 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 2041 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa 2101 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 2161 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga 2221 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 2281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 2341 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 2401 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 2461 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 2521 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga 2581 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 2641 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 2701 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 2761 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 2821 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 2881 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 2941 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 3001 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 3061 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 3121 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 3181 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 3241 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 3301 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 3361 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 3421 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 3481 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 3541 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag 3601 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt 3661 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc 3721 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc 3781 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata 3841 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 3901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 3961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 4021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 4081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 4141 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 4201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 4261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 4321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 4381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 4441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 4501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 4561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 4621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 4681 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatt //