LOCUS Exported 5220 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses human cofilin fused to EGFP in mammalian cells . ACCESSION . VERSION . KEYWORDS pEGFP-N1 human cofilin WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5220) AUTHORS Garvalov BK, Flynn KC, Neukirchen D, Meyn L, Teusch N, Wu X, Brakebusch C, Bamburg JR, Bradke F TITLE Cdc42 regulates cofilin during the establishment of neuronal polarity. JOURNAL J Neurosci. 2007 Nov 28;27(48):13117-29. PUBMED 18045906 REFERENCE 2 (bases 1 to 5220) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5220 /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 1160..1169 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1166..1885 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1166..1882 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1211..1232) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1472..1491) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1819..1840 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2008..2129 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2045..2064) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2099..2118 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2136..2591) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2273..2294) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2485..2504 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2618..2722 /gene="bla" /label=AmpR promoter promoter 2724..3081 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2747..2767) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2932..3067 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2994..3013 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3116..3910 /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(3170..3189) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3780..3799 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4098..4117) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4142..4189 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4518..5106 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5007..5026 /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 attcaccacc atggcctccg gtgtggctgt 661 ctctgatggt gtcatcaagg tgttcaacga catgaaggtg cgtaagtctt caacgccaga 721 ggaggtgaag aagcgcaaga aggcggtgct cttctgcctg agtgaggaca agaagaacat 781 catcctggag gagggcaagg agatcctggt gggcgatgtg ggccagactg tcgacgatcc 841 ctacgccacc tttgtcaaga tgctgccaga taaggactgc cgctatgccc tctatgatgc 901 aacctatgag accaaggaga gcaagaagga ggatctggtg tttatcttct gggcccccga 961 gtctgcgccc cttaagagca aaatgattta tgccagctcc aaggacgcca tcaagaagaa 1021 gctgacaggg atcaagcatg aattgcaagc aaactgctac gaggaggtca aggaccgctg 1081 caccctggca gagaagctgg ggggcagtgc ggtcatctcc ctggagggca agcctttgtt 1141 atcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 1201 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 1261 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 1321 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 1381 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 1441 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 1501 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 1561 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 1621 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 1681 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 1741 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 1801 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 1861 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc 1921 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa 1981 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa 2041 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 2101 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt 2161 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg 2221 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg 2281 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta 2341 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg 2401 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa 2461 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct 2521 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct 2581 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 2641 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 2701 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag 2761 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 2821 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 2881 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 2941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 3001 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 3061 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat 3121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 3181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 3241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga 3301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 3361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 3421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 3481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 3541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 3601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga 3661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 3721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 3781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 3841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 3901 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 3961 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4021 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4081 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg 4141 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg 4201 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac 4261 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc 4321 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag 4381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 4441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 4501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 4681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 4981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 5041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 5101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 5161 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat //