LOCUS Exported 5855 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION golden gate entry vector for mCherry-LEXY tagging of proteins (no constitutive NLS). ACCESSION . VERSION . KEYWORDS pLEXY (pDN137) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5855) AUTHORS Niopek D, Wehler P, Roensch J, Eils R, Di Ventura B TITLE Optogenetic control of nuclear protein export. JOURNAL Nat Commun. 2016 Feb 8;7:10624. doi: 10.1038/ncomms10624. PUBMED 26853913 REFERENCE 2 (bases 1 to 5855) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5855 /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" CDS 993..1298 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 1177..1196 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 1335..2039 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(1477..1495) /label=mCherry-R /note="mCherry, reverse primer" polyA_signal 2643..2764 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2680..2699) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2734..2753 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2771..3226) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2908..2929) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3120..3139 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3253..3357 /gene="bla" /label=AmpR promoter promoter 3359..3716 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3382..3402) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3567..3702 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3629..3648 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3751..4545 /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(3805..3824) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4415..4434 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4733..4752) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4777..4824 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5153..5741 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5642..5661 /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 ccggactcag atctcgagct caagcttcga attccgccac catgaagtct tccgcgtgga 661 tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata 721 agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc 781 gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa 841 tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg 901 ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg 961 ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa 1021 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc 1081 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac 1141 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca 1201 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca 1261 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca 1321 gccagtgaag acaagtgagc aagggcgagg aggataacat ggccatcatc aaggagttca 1381 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 1441 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg 1501 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct 1561 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca 1621 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 1681 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg 1741 acggccccgt aatgcagaag aaaaccatgg gctgggaggc ctcctccgag cggatgtacc 1801 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc 1861 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg 1921 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg 1981 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg 2041 gtggatctgg aggttcaggt ggaagtttgg ctactacact tgaacgtatt gagaagaact 2101 ttgtcattac tgacccaaga ttgccagata atcccattat attcgcgtcc gatagtttct 2161 tgcagttgac agaatatagc cgtgaagaaa ttttgggaag aaactgcagg tttctacaag 2221 gtcctgaaac tgatcgcgcg acagtgagaa aaattagaga tgccatagat aaccaaacag 2281 aggtcactgt tcagctgatt aattatacaa agagtggtaa aaagttctgg aacctctttc 2341 acttgcagcc tatgcgagat cagaagggag atgtccagta ctttattggg gttcagttgg 2401 atggaactga gcatgtccga gatgctgccg agagagaggg agtcatgctg attaagaaaa 2461 ctgcagaaaa tattgacgag ctgctgaagg agctggccga cctgaacctg gactagctcg 2521 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt 2581 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg 2641 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 2701 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 2761 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa 2821 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa 2881 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac 2941 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa 3001 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct 3061 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa 3121 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc 3181 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt 3241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 3301 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct 3361 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3421 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3481 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3541 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3601 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 3661 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc 3721 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt 3781 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 3841 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 3901 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg 3961 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 4021 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 4081 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 4141 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 4201 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 4261 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 4321 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 4381 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 4441 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 4501 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 4561 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 4621 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 4681 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac 4741 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 4801 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg 4861 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac 4921 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc 4981 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat 5041 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 5101 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 5161 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 5221 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 5281 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact 5341 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 5401 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 5461 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 5521 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 5581 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 5641 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 5701 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 5761 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 5821 ctgattctgt ggataaccgt attaccgcca tgcat //