LOCUS Exported 5721 bp ds-DNA circular SYN 16-AUG-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEYFP(C1)-PP1alpha SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5721) AUTHORS Trinkle-Mulcahy L, Sleeman JE, Lamond AI TITLE Dynamic targeting of protein phosphatase 1 within the nuclei of living mammalian cells. JOURNAL J Cell Sci. 2001 Dec;114(Pt 23):4219-28. PUBMED 11739654 REFERENCE 2 (bases 1 to 5721) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5721 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 100..403 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 404..607 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 558..578 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 646..655 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 652..1368 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(697..718) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(958..977) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1305..1326 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2548..2669 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2585..2604) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2639..2658 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2676..3131) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2813..2834) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3025..3044 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3158..3262 /gene="bla" /label=AmpR promoter promoter 3264..3621 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3287..3307) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3472..3607 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3534..3553 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3656..4450 /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(3710..3729) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4320..4339 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4638..4657) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4682..4729 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5058..5646 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5547..5566 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat 61 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa 121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt 181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta 241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt 301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc 361 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca 421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat 481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa 541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag 601 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc 661 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta 721 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg 781 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc 841 accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac 901 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac 961 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc 1021 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag 1081 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag 1141 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac 1201 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc 1261 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag 1321 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga 1381 tctcgagctc aagcttcgaa ttcgatgtcc gacagcgaga agctcaacct ggactcgatc 1441 atcgggcgcc tgctggaagt gcagggctcg cggcctggca agaatgtaca gctgacagag 1501 aacgagatcc gcggtctgtg cctgaaatcc cgggagattt ttctgagcca gcccattctt 1561 ctggagctgg aggcacccct caagatctgc ggtgacatac acggccagta ctacgacctt 1621 ctgcgactat ttgagtatgg cggtttccct cccgagagca actacctctt tctgggggac 1681 tatgtggaca ggggcaagca gtccttggag accatctgcc tgctgctggc ctataagatc 1741 aagtaccccg agaacttctt cctgctccgt gggaaccacg agtgtgccag catcaaccgc 1801 atctatggtt tctacgatga gtgcaagaga cgctacaaca tcaaactgtg gaaaaccttc 1861 actgactgct tcaactgcct gcccatcgcg gccatagtgg acgaaaagat cttctgctgc 1921 cacggaggcc tgtccccgga cctgcagtct atggagcaga ttcggcggat catgcggccc 1981 acagatgtgc ctgaccaggg cctgctgtgt gacctgctgt ggtctgaccc tgacaaggac 2041 gtgcagggct ggggcgagaa cgaccgtggc gtctctttta cctttggagc cgaggtggtg 2101 gccaagttcc tccacaagca cgacttggac ctcatctgcc gagcacacca ggtggtagaa 2161 gacggctacg agttctttgc caagcggcag ctggtgacac ttttctcagc tcccaactac 2221 tgtggcgagt ttgacaatgc tggcgccatg atgagtgtgg acgagaccct catgtgctct 2281 ttccagatcc tcaagcccgc cgacaagaac aaggggaagt acgggcagtt cagtggcctg 2341 aaccctggag gccgacccat caccccaccc cgcaattccg ccaaagccaa gaaataggtc 2401 gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc 2461 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa 2521 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa 2581 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 2641 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt 2701 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg 2761 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg 2821 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta 2881 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg 2941 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa 3001 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct 3061 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct 3121 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 3181 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 3241 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag 3301 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 3361 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3421 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 3481 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 3541 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 3601 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat 3661 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 3721 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 3781 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga 3841 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 3901 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 3961 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 4021 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 4081 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 4141 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga 4201 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 4261 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 4321 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 4381 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 4441 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 4501 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4561 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4621 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg 4681 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg 4741 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac 4801 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc 4861 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag 4921 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 4981 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 5041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 5101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 5161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 5221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 5281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 5341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 5401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 5461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 5521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 5581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 5641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 5701 cacatgttct ttcctgcgtt a //