LOCUS Exported 5979 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEGFP-N1-ACTR3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5979) AUTHORS Welch MD, DePace AH, Verma S, Iwamatsu A, Mitchison TJ TITLE The human Arp2/3 complex is composed of evolutionarily conserved subunits and is localized to cellular regions of dynamic actin filament assembly. JOURNAL J Cell Biol 1997 Jul 28;138(2):375-84. PUBMED 9230079 REFERENCE 2 (bases 1 to 5979) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5979 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1859..1868 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1865..2584 /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 1865..2581 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1910..1931) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2171..2190) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2518..2539 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2707..2828 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2744..2763) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2798..2817 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2835..3290) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2972..2993) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3184..3203 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3317..3421 /gene="bla" /label=AmpR promoter promoter 3423..3780 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3446..3466) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3631..3766 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3693..3712 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3815..4609 /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(3869..3888) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4479..4498 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4797..4816) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4841..4888 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5217..5805 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5706..5725 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggactcag atctcgagct caagcttcga attcaggcga cggaggaaga tggcgggacg 601 gctgccggcc tgtgtggtgg actgtggcac ggggtataca aaactaggat atgctggaaa 661 tacagaacca cagtttatca tcccttcctg tattgctatt aaggagtcag caaaagtggg 721 tgatcaagct caaaggaggg tgatgaaagg tgttgatgac ctagacttct tcattggtga 781 tgaagcaata gaaaaaccta catatgcaac aaagtggcca atccgccatg gtatagttga 841 agattgggac ttaatggaaa ggtttatgga gcaagtgatc tttaaatatt taagggcaga 901 acctgaagac cattattttc ttttgactga acctccattg aatactccag aaaacaggga 961 atatactgct gaaataatgt ttgagtcctt caatgttcca ggcttgtaca ttgctgtgca 1021 ggctgttctt gccttagctg catcttggac ctcaagacaa gtaggagaac ggacgttgac 1081 cggtacggta atagacagtg gagatggtgt cactcatgtc attcctgtgg ctgaagggta 1141 tgtgattggc agctgtatta aacacattcc aatcgcagga cgagatataa catattttat 1201 tcagcaactg ctgagagacc gagaagtagg aatccctcca gaacaatcct tggaaactgc 1261 taaggcagta aaggagcgct atagttatgt ctgcccagat ttagtaaaag aatttaacaa 1321 gtatgataca gatgggtcaa aatggattaa acagtatact ggaatcaatg ctatctcaaa 1381 gaaagagttt tctatcgatg ttggttatga gagatttttg ggacctgaaa tcttttttca 1441 tccagagttt gctaatccag actttacaca acctatctca gaagttgtag atgaagtaat 1501 tcagaattgt cctattgatg tcagacgtcc tctctacaag aatattgtcc tctctggagg 1561 ttcaaccatg ttcagggact ttggacgtcg cttgcaaaga gatttgaaaa gaactgtaga 1621 tgcccggctg aaattaagtg aggaattgag tggtggtaga ttgaagccaa aacctattga 1681 tgtacaagtc attacacacc acatgcagcg atatgcagtt tggtttggag gatcaatgct 1741 ggcttccacg cctgagttct accaagtatg ccacaccaaa aaggattatg aagaaattgg 1801 acctagcatt tgtcgtcaca atccagtgtt tggagtcatg tcccgggatc caccggtcgc 1861 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct 1921 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac 1981 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc 2041 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat 2101 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat 2161 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac 2221 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg 2281 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa 2341 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct 2401 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa 2461 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat 2521 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa 2581 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt 2641 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 2701 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 2761 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 2821 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 2881 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 2941 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 3001 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 3061 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 3121 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 3181 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 3241 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 3301 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 3361 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 3421 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 3481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3541 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3601 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3661 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 3721 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 3781 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 3841 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 3901 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 3961 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 4021 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 4081 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 4141 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 4201 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 4261 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 4321 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 4381 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 4441 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 4501 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 4561 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 4621 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4681 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4741 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 4801 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 4861 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 4921 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 4981 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 5041 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 5101 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 5161 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 5221 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 5281 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 5341 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 5401 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 5461 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 5521 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 5581 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5641 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5701 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5761 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 5821 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 5881 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat 5941 tacggggtca ttagttcata gcccatatat ggagttccg //