LOCUS Exported 7884 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION DULIP negative control (prey) to exclude unspecific background binding. ACCESSION . VERSION . KEYWORDS FL-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7884) AUTHORS Trepte P, Buntru A, Klockmeier K, Willmore L, Arumughan A, Secker C, Zenkner M, Brusendorf L, Rau K, Redel A, Wanker EE TITLE DULIP: A Dual Luminescence-Based Co-Immunoprecipitation Assay for Interactome Mapping in Mammalian Cells. JOURNAL J Mol Biol. 2015 Oct 23;427(21):3375-88. doi: 10.1016/j.jmb.2015.08.003. Epub 2015 Aug 8. PUBMED 26264872 REFERENCE 2 (bases 1 to 7884) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7884 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 940..949 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 946..2595 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(1012..1030) /label=LucNrev /note="Firefly luciferase, reverse primer" CDS 2620..2661 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" protein_bind 2674..2698 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2701..3408 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(2846..2864) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3114..3133 /label=mCherry-F /note="mCherry, forward primer" protein_bind complement(3410..3434) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(3480..3497) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3486..3710 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3756..4184 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3843..3862) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4053..4074 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4193..4213) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4198..4527 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4378..4513 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4440..4459 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4594..5388 /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(4648..4667) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5258..5277 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5562..5683 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5599..5618) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5653..5672 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5732..5748) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5732..5748) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5745..5767) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5756..5772 /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(5780..5810) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5825..5846 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5963..5980) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6134..6719) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6214..6233) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6890..7750) /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 7513..7532 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7751..7855) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta 901 gcgtttaaac ttaagcttgg taccgagctc ggatccgccg ccaccatgga agacgccaaa 961 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa 1021 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca 1081 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa 1141 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct 1201 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg 1261 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg 1321 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc 1381 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac 1441 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc 1501 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct 1561 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct 1621 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac 1681 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg 1741 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg 1801 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat 1861 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa 1921 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact 1981 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt 2041 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat 2101 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg 2161 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac 2221 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa 2281 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc 2341 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt 2401 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa 2461 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt 2521 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc 2581 ggaaagatcg ccgtggacgc gaattccgcg gggctcgacg gtaagcctat ccctaaccct 2641 ctcctcggtc tcgattctac gctcgaggat atcacaagtt tgtacaaaaa agcaggcttc 2701 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag 2761 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc 2821 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc 2881 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac 2941 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc 3001 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac 3061 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta 3121 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc 3181 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct 3241 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc 3301 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa 3361 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagga cccagctttc 3421 ttgtacaaag tggtgatatc gcggccgctc tagagggccc gtttaaaccc gctgatcagc 3481 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3541 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3601 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3661 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3721 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3781 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3841 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3901 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 3961 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4021 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4081 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 4141 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 4201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4381 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4441 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4501 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4561 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4621 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 4681 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4741 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 4801 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 4861 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 4921 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 4981 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 5041 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 5101 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 5161 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 5221 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 5281 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 5341 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 5401 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 5461 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5521 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5581 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5641 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5701 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5761 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5821 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5881 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5941 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 6001 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 6061 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 6121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 6181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 6241 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6301 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6361 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6421 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6481 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6541 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6601 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6661 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6721 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6781 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6841 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 6901 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 6961 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 7021 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 7081 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 7141 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 7201 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 7261 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7321 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7381 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7441 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7501 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7561 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7621 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 7681 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 7741 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 7801 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 7861 ttccccgaaa agtgccacct gacg //