LOCUS Exported 6768 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION MAC-tagged gene expression. ACCESSION . VERSION . KEYWORDS MAC_C_TOM20 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6768) AUTHORS Liu X, Salokas K, Tamene F, Jiu Y, Weldatsadik RG, Ohman T, Varjosalo M TITLE An AP-MS- and BioID-compatible MAC-tag enables comprehensive mapping of protein interactions and subcellular localizations. JOURNAL Nat Commun. 2018 Mar 22;9(1):1188. doi: 10.1038/s41467-018-03523-2. PUBMED 29568061 REFERENCE 2 (bases 1 to 6768) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6768 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 16..395 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 396..599 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 550..570 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 601..619 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 622..640 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 650..674 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 817..841 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(1286..1310) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1357..1380 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 1417..1440 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 1471..1497 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1498..2457 /codon_start=1 /product="promiscuous R118G mutant of the E. coli biotin protein ligase (Cronan, 2005)" /label=BirA(R118G) /note="also known as BirA* or BioID" /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK" primer_bind complement(2501..2518) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2507..2731 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2864..2883) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3015..3062 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 3070..4092 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4222..4343 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4259..4278) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4313..4332 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4392..4408) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4392..4408) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4405..4427) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4416..4432 /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(4440..4470) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4485..4506 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4623..4640) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4794..5382) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4874..4893) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5553..6413) /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 6176..6195 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6414..6518) /gene="bla" /label=AmpR promoter primer_bind complement(6593..6612) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt 61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca 421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc 601 tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga gctcgtttag 661 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc 721 gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc gagctcggat 781 ccactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa aaaagcaggc 841 tccaccatgg tgggtcggaa cagcgccatc gccgccggtg tatgcggggc ccttttcatt 901 gggtactgca tctacatcga ccgcaaaaga cgaagtgacc ccaacttcaa gaacaggctt 961 cgagaacgaa gaaagaaaca gaagcttgcc aaggagagag ctgggctttc caagttacct 1021 gaccttaaag atgctgaagc tgttcagaag ttcttccttg aagaaataca gcttggtgaa 1081 gagttactag ctcaaggtga atatgagaag ggcgtagacc atctgacaaa tgcaattgct 1141 gtgtgtggac agccacagca gttactgcag gtcttacagc aaactcttcc accaccagtg 1201 ttccagatgc ttctgactaa gctcccaaca attagtcaga gaattgtaag tgctcagagc 1261 ttggctgaag atgatgtgga attggaccca gctttcttgt acaaagtggt tgatatcaag 1321 gccgccgaca tcaccagcct gtacaagaaa gtggggtgga gccatcccca gttcgagaag 1381 ggcggaggat ctggcggcgg aagcggcgga ggaagctggt cccaccctca gtttgagaaa 1441 ggcaccgagc tgggcagcac catggccagc tacccctacg acgtgcccga ctacgccaag 1501 gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca acggcgagtt ccactctggc 1561 gagcagctgg gcgagacact gggcatgagc agagccgcca tcaacaagca catccagacc 1621 ctgcgggact ggggcgtgga cgtgttcaca gtgcccggca agggctacag cctgcccgag 1681 cccatccagc tgctgaacgc caagcagatc ctgggccagc tggatggcgg atctgtggcc 1741 gtgctgcccg tgatcgacag caccaaccag tatctgctcg accggatcgg cgagctgaag 1801 tccggcgacg cctgtatcgc cgagtaccag caggccggca gaggcggcag gggcagaaag 1861 tggttcagcc ccttcggcgc caacctgtac ctgagcatgt tctggagact ggaacagggc 1921 ccagccgccg ctattggcct gagcctggtc atcggcatcg tgatggccga ggtgctgcgg 1981 aagctgggcg ccgacaaagt gcgcgtgaag tggcccaacg acctgtatct gcaggaccgg 2041 aagctggccg gcatcctggt ggaactgacc ggcaaaacag gcgacgccgc ccagatcgtg 2101 atcggcgctg gcatcaacat ggccatgcgg cgggtggaag agagcgtggt caaccagggc 2161 tggatcaccc tgcaggaagc cggcatcaac ctggaccgga acaccctggc cgccatgctg 2221 atcagagagc tgagagccgc cctggaactg ttcgagcagg aaggcctggc cccctacctg 2281 agcagatggg agaagctgga caacttcatc aaccggcccg tgaaactgat catcggcgac 2341 aaagagatct tcggcatcag ccggggcatc gacaagcagg gggccctgct cctggaacag 2401 gacggcatca tcaagccctg gatgggcggc gagatcagcc tgcggagcgc cgagaagtga 2461 tgactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt 2521 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 2581 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 2641 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 2701 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 2761 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2821 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2881 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 2941 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3001 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact 3061 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag 3121 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc 3181 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac 3241 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt 3301 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc 3361 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc 3421 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg 3481 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat 3541 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc 3601 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat 3661 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc 3721 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg 3781 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga 3841 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg 3901 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga 3961 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc 4021 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg 4081 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg 4141 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4201 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 4261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4321 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 4381 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 4441 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 4501 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 4561 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 4621 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 4681 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 4741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 4801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 4861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 4921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 4981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 5041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 5101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 5161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 5221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 5281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 5341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 5401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 5461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 5521 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 5581 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 5641 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 5701 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 5761 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 5821 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 5881 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 5941 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6001 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 6061 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 6121 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 6181 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 6241 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6301 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 6361 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 6421 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 6481 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 6541 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 6601 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 6661 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 6721 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacggg //