LOCUS Exported 5792 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of N-terminal fusion of mClover3 to actin. ACCESSION . VERSION . KEYWORDS pKanCMV-mClover3-18aa-actin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5792) AUTHORS Bajar BT, Wang ES, Lam AJ, Kim BB, Jacobs CL, Howe ES, Davidson MW, Lin MZ, Chu J TITLE Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting. JOURNAL Sci Rep. 2016 Feb 16;6:20889. doi: 10.1038/srep20889. PUBMED 26879144 REFERENCE 2 (bases 1 to 5792) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5792 /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 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2638..2759 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2675..2694) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2729..2748 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2766..3221) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2903..2924) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3115..3134 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3248..3352 /gene="bla" /label=AmpR promoter promoter 3354..3711 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3377..3397) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3562..3697 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3624..3643 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3746..4540 /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(3800..3819) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4410..4429 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4728..4747) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4772..4819 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5148..5736 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5637..5656 /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 ccggtcgcca ccatggtgtc taagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga gggcgagggc 721 gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaacttc aacagccact acgtctatat cacggccgac 1081 aagcagaaga actgcatcaa ggctaacttc aagatccgcc acaacgttga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag ccatcagtcc aagctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga ttacacatgg catggacgag 1321 ctgtacaagt ccggactcag atctggcagc ggtggaggca gcgcatccgg cggaagcgga 1381 agcgatgatg atatcgccgc gctcgtcgtc gacaacggct ccggcatgtg caaggccggc 1441 ttcgcgggcg acgatgcccc ccgggccgtc ttcccctcca tcgtggggcg ccccaggcac 1501 cagggcgtga tggtgggcat gggtcagaag gattcctatg tgggcgacga ggcccagagc 1561 aagagaggca tcctcaccct gaagtacccc atcgagcacg gcatcgtcac caactgggac 1621 gacatggaga aaatctggca ccacaccttc tacaatgagc tgcgtgtggc tcccgaggag 1681 caccccgtgc tgctgaccga ggcccccctg aaccccaagg ccaaccgcga gaagatgacc 1741 cagatcatgt ttgagacctt caacacccca gccatgtacg ttgctatcca ggctgtgcta 1801 tccctgtacg cctctggccg taccactggc atcgtgatgg actccggtga cggggtcacc 1861 cacactgtgc ccatctacga ggggtatgcc ctcccccatg ccatcctgcg tctggacctg 1921 gctggccggg acctgactga ctacctcatg aagatcctca ccgagcgcgg ctacagcttc 1981 accaccacgg ccgagcggga aatcgtgcgt gacattaagg agaagctgtg ctacgtcgcc 2041 ctggacttcg agcaagagat ggccacggct gcttccagct cctccctgga gaagagctac 2101 gagctgcctg acggccaggt catcaccatt ggcaatgagc ggttccgctg ccctgaggca 2161 ctcttccagc cttccttcct gggcatggag tcctgtggca tccacgaaac taccttcaac 2221 tccatcatga agtgtgacgt ggacatccgc aaagacctgt acgccaacac agtgctgtct 2281 ggcggcacca ccatgtaccc tggcattgcc gacaggatgc agaaggagat cactgccctg 2341 gcacccagca caatgaagat caagatcatt gctcctcctg agcgcaagta ctccgtgtgg 2401 atcggcggct ccatcctggc ctcgctgtcc accttccagc agatgtggat cagcaagcag 2461 gagtatgacg agtccggccc ctccatcgtc caccgcaaat gcttctaggg atccaccgga 2521 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa 2581 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac 2641 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 2701 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa 2761 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 2821 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 2881 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 2941 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 3001 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 3061 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 3121 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 3181 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg 3241 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 3301 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc 3361 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 3421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 3481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 3541 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 3601 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 3661 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca 3721 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 3781 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 3841 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 3901 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac 3961 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 4021 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 4081 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 4141 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 4201 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 4261 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 4321 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 4381 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 4441 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 4501 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 4561 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 4621 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 4681 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag 4741 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 4801 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 4861 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 4921 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc 4981 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 5041 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 5101 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 5161 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 5221 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 5281 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 5341 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 5401 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 5461 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 5521 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 5581 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 5641 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 5701 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 5761 cggttcctgg ccttttgctg gccttttgct ca //