LOCUS Exported 5220 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses psuedo-phosporylated, non-activatable cofilin fused to EGFP in mammalian cells . ACCESSION . VERSION . KEYWORDS pEGFP-N1 human cofilin S3E 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 16..319 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 320..523 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 474..494 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1115..1124 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1121..1840 /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 1121..1837 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1166..1187) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1427..1446) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1774..1795 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1963..2084 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2000..2019) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2054..2073 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2091..2546) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2228..2249) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2440..2459 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2573..2677 /gene="bla" /label=AmpR promoter promoter 2679..3036 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2702..2722) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2887..3022 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2949..2968 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3071..3865 /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(3125..3144) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3735..3754 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4053..4072) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4097..4144 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4473..5061 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4962..4981 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca 541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattca ccaccatggc 601 cgaaggtgtg gctgtctctg atggtgtcat caaggtgttc aacgacatga aggtgcgtaa 661 gtcttcaacg ccagaggagg tgaagaagcg caagaaggcg gtgctcttct gcctgagtga 721 ggacaagaag aacatcatcc tggaggaggg caaggagatc ctggtgggcg atgtgggcca 781 gactgtcgac gatccctacg ccacctttgt caagatgctg ccagataagg actgccgcta 841 tgccctctat gatgcaacct atgagaccaa ggagagcaag aaggaggatc tggtgtttat 901 cttctgggcc cccgagtctg cgccccttaa gagcaaaatg atttatgcca gctccaagga 961 cgccatcaag aagaagctga cagggatcaa gcatgaattg caagcaaact gctacgagga 1021 ggtcaaggac cgctgcaccc tggcagagaa gctggggggc agtgcggtca tctccctgga 1081 gggcaagcct ttgttatccc gggatccacc ggtcgccacc atggtgagca agggcgagga 1141 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa 1201 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt 1261 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta 1321 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc 1381 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta 1441 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa 1501 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa 1561 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa 1621 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac 1681 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc 1741 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc 1801 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc 1861 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc 1921 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct 1981 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 2041 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag 2101 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca 2161 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag 2221 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg 2281 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt 2341 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag 2401 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc 2461 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc 2521 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac 2581 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 2641 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg 2701 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2761 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 2821 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 2881 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 2941 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag 3001 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg 3061 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 3121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 3181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 3241 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 3301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 3361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 3421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 3481 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga 3541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag 3601 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat 3661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 3721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 3781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 3841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg 3901 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc 3961 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 4021 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa 4081 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt 4141 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 4201 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 4261 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac 4321 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 4381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 4441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 4501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 4561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt 4621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 4681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 4741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 4801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 4861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 4921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 4981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 5041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 5101 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt 5161 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc //