LOCUS Exported 7231 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express Ran T24N mutant in mammalian cells (CMV promoter). Also suitable for in vitro transcription (T7 promoter) for injecting mRNA into oocytes.. ACCESSION . VERSION . KEYWORDS pcDNA-Ran-T24N-mRFP1-polyA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7231) AUTHORS Inoue A, Zhang Y TITLE Nucleosome assembly is required for nuclear pore complex assembly in mouse zygotes. JOURNAL Nat Struct Mol Biol. 2014 Jul;21(7):609-16. doi: 10.1038/nsmb.2839. Epub 2014 Jun 8. PUBMED 24908396 REFERENCE 2 (bases 1 to 7231) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7231 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 277..306 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 322..339 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(362..379) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 368..592 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 638..1066 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(725..744) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 935..956 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1075..1095) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1080..1409 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1260..1395 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1322..1341 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 1476..2267 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1530..1549) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2140..2159 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 2446..2567 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2483..2502) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2537..2556 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(2616..2632) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2616..2632) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2629..2651) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2640..2656 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2664..2694) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2709..2730 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2847..2864) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3018..3606) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3098..3117) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3777..4637) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4400..4419 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4638..4742) /gene="bla" /label=AmpR promoter primer_bind complement(4817..4836) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 5008..5387 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5388..5591 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5542..5562 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5636..5655 /label=T7 /note="T7 promoter, forward primer" promoter 5636..5654 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 6335..7012 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" primer_bind complement(6465..6483) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 6733..6752 /label=mCherry-F /note="mCherry, forward primer" primer_bind 6927..6950 /label=DsRed1-C /note="DsRed1, forward primer" ORIGIN 1 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 61 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 121 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 181 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa atacgcatca 241 tacgctgtgg cgtacctcga gtctagaggg cccttcgaac aaaaactcat ctcagaagag 301 gatctgaata tgcataccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca 361 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 421 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 481 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 541 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 601 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 661 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 721 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 781 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 841 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 901 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 961 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 1021 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 1081 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 1141 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 1201 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 1261 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 1321 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 1381 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg 1441 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 1501 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 1561 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 1621 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt 1681 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 1741 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 1801 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 1861 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 1921 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 1981 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg 2041 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 2101 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 2161 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 2221 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctta gcgggactct 2281 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 2341 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 2401 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 2461 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 2521 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 2581 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 2641 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 2701 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 2761 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 2821 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 2881 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 2941 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 3001 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 3061 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 3121 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 3181 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 3241 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 3301 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 3361 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 3421 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 3481 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 3541 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 3601 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 3661 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 3721 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 3781 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 3841 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 3901 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 3961 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 4021 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 4081 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 4141 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 4201 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 4261 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 4321 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 4381 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 4441 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 4501 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 4561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 4621 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 4681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 4741 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc 4801 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc 4861 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc 4921 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc 4981 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat 5041 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 5101 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 5161 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 5221 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 5281 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 5341 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 5401 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 5461 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 5521 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 5581 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata 5641 cgactcacta tagggagacc caagctggct agttaagctt atggccgccc agggagagcc 5701 gcaggtccag ttcaagctcg tcctggtggg cgacggcggc accgggaaga acaccttcgt 5761 gaagcgccac ttgacgggcg agtttgagaa gaagtatgta gccaccctgg gcgtggaggt 5821 gcacccgctc gtcttccata ccaacagagg acccatcaag ttcaacgtgt gggacacggc 5881 cggccaggag aagttcgggg gcctgcgcga tggctactac atccaagccc agtgtgccat 5941 tataatgttt gatgtaacct caagagttac ttacaagaat gtacctaact ggcatagaga 6001 tctggtacga gtgtgtgaaa acatccccat tgtattgtgt ggcaacaaag tggatattaa 6061 agacaggaaa gtgaaggcaa aatctattgt cttccaccgg aagaagaatc ttcagtacta 6121 tgacatttct gccaaaagta actacaactt tgaaaagcct ttcctctggc ttgccagaaa 6181 gctcattgga gatcctaact tggagtttgt tgccatgcct gctcttgccc cacctgaggt 6241 ggtcatggac ccagctttgg cagcacagta cgagcatgat ttagaggttg ctcagacgac 6301 tgctctccca gatgaggatg atgacctgga tccaatggcc tcctccgagg acgtcatcaa 6361 ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat 6421 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac 6481 caagggcggc cccctgccct tcgcctggga catcctgtcc cctcagttcc agtacggctc 6541 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga 6601 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca 6661 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt 6721 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg 6781 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag atgaggctga agctgaagga 6841 cggcggccac tacgacgccg aggtcaagac cacctacatg gccaagaagc ccgtgcagct 6901 gcccggcgcc tacaagaccg acatcaagct ggacatcacc tcccacaacg aggactacac 6961 catcgtggaa cagtacgagc gcgccgaggg ccgccactcc accggcgcct aagcggccgc 7021 tcgactatgg gcaccaaaga acctgtaaac gttatctttt taaattgaat gtgcacaaat 7081 aaaagtttgg aaaagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7141 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7201 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa a //