LOCUS Exported 8052 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION MAC-tagged gene expression. ACCESSION . VERSION . KEYWORDS MAC_C_CH60 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8052) 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 8052) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8052 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 32..411 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 412..615 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 566..586 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 617..635 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 638..656 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 666..690 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 833..857 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2586..2610) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2657..2680 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2717..2740 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2771..2797 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2798..3757 /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(3801..3818) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3807..4031 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4164..4183) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 4315..4362 /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 4370..5392 /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 5522..5643 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5559..5578) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5613..5632 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5692..5708) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5692..5708) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5705..5727) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5716..5732 /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(5740..5770) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5785..5806 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5923..5940) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6094..6682) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6174..6193) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6853..7713) /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 7476..7495 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7714..7818) /gene="bla" /label=AmpR promoter primer_bind complement(7893..7912) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag 61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg 421 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc 481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa 541 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 601 tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga tagagatcgt 661 cgacgagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac 721 ctccatagaa gacaccggga ccgatccagc ctccggactc tagcgtttaa acttaagctt 781 ggtaccgagc tcggatccac tagtccagtg tggtggaatt ctgcagatat caacaagttt 841 gtacaaaaaa gcaggctcca ccatgcttcg gttacccaca gtctttcggc agatgagacc 901 ggtgtccagg gtactggctc ctcatctcac ccgggcttat gccaaagatg taaaatttgg 961 tgcagatgcc cgagccttaa tgcttcaagg tgtagacctt ttagccgatg ctgtggccgt 1021 tacaatgggg ccaaagggaa gaacagtgat tattgagcag agttggggaa gtcccaaagt 1081 aacaaaagat ggtgtgactg ttgcaaagtc aattgactta aaagataaat acaagaacat 1141 tggagctaaa cttgttcaag atgttgccaa taacacaaat gaagaagctg gggatggcac 1201 taccactgct actgtactgg cacgctctat agccaaggaa ggcttcgaga agattagcaa 1261 aggtgctaat ccagtggaaa tcaggagagg tgtgatgtta gctgttgatg ctgtaattgc 1321 tgaacttaaa aagcagtcta aacctgtgac cacccctgaa gaaattgcac aggttgctac 1381 gatttctgca aacggagaca aagaaattgg caatatcatc tctgatgcaa tgaaaaaagt 1441 tggaagaaag ggtgtcatca cagtaaagga tggaaaaaca ctgaatgatg aattagaaat 1501 tattgaaggc atgaagtttg atcgaggcta tatttctcca tactttatta atacatcaaa 1561 aggtcagaaa tgtgaattcc aggatgccta tgttctgttg agtgaaaaga aaatttctag 1621 tatccagtcc attgtacctg ctcttgaaat tgccaatgct caccgtaagc ctttggtcat 1681 aatcgctgaa gatgttgatg gagaagctct aagtacactc gtcttgaata ggctaaaggt 1741 tggtcttcag gttgtggcag tcaaggctcc agggtttggt gacaatagaa agaaccagct 1801 taaagatatg gctattgcta ctggtggtgc agtgtttgga gaagagggat tgaccctgaa 1861 tcttgaagac gttcagcctc atgacttagg aaaagttgga gaggtcattg tgaccaaaga 1921 cgatgccatg ctcttaaaag gaaaaggtga caaggctcaa attgaaaaac gtattcaaga 1981 aatcattgag cagttagatg tcacaactag tgaatatgaa aaggaaaaac tgaatgaacg 2041 gcttgcaaaa ctttcagatg gagtggctgt gctgaaggtt ggtgggacaa gtgatgttga 2101 agtgaatgaa aagaaagaca gagttacaga tgcccttaat gctacaagag ctgctgttga 2161 agaaggcatt gttttgggag ggggttgtgc cctccttcga tgcattccag ccttggactc 2221 attgactcca gctaatgaag atcaaaaaat tggtatagaa attattaaaa gaacactcaa 2281 aattccagca atgaccattg ctaagaatgc aggtgttgaa ggatctttga tagttgagaa 2341 aattatgcaa agttcctcag aagttggtta tgatgctatg gctggagatt ttgtgaatat 2401 ggtggaaaaa ggaatcattg acccaacaaa ggttgtgaga actgctttat tggatgctgc 2461 tggtgtggcc tctctgttaa ctacagcaga agttgtagtc acagaaattc ctaaagaaga 2521 gaaggaccct ggaatgggtg caatgggtgg aatgggaggt ggtatgggag gtggcatgtt 2581 cttggaccca gctttcttgt acaaagtggt tgatatcaag gccgccgaca tcaccagcct 2641 gtacaagaaa gtggggtgga gccatcccca gttcgagaag ggcggaggat ctggcggcgg 2701 aagcggcgga ggaagctggt cccaccctca gtttgagaaa ggcaccgagc tgggcagcac 2761 catggccagc tacccctacg acgtgcccga ctacgccaag gacaacaccg tgcccctgaa 2821 gctgatcgcc ctgctggcca acggcgagtt ccactctggc gagcagctgg gcgagacact 2881 gggcatgagc agagccgcca tcaacaagca catccagacc ctgcgggact ggggcgtgga 2941 cgtgttcaca gtgcccggca agggctacag cctgcccgag cccatccagc tgctgaacgc 3001 caagcagatc ctgggccagc tggatggcgg atctgtggcc gtgctgcccg tgatcgacag 3061 caccaaccag tatctgctcg accggatcgg cgagctgaag tccggcgacg cctgtatcgc 3121 cgagtaccag caggccggca gaggcggcag gggcagaaag tggttcagcc ccttcggcgc 3181 caacctgtac ctgagcatgt tctggagact ggaacagggc ccagccgccg ctattggcct 3241 gagcctggtc atcggcatcg tgatggccga ggtgctgcgg aagctgggcg ccgacaaagt 3301 gcgcgtgaag tggcccaacg acctgtatct gcaggaccgg aagctggccg gcatcctggt 3361 ggaactgacc ggcaaaacag gcgacgccgc ccagatcgtg atcggcgctg gcatcaacat 3421 ggccatgcgg cgggtggaag agagcgtggt caaccagggc tggatcaccc tgcaggaagc 3481 cggcatcaac ctggaccgga acaccctggc cgccatgctg atcagagagc tgagagccgc 3541 cctggaactg ttcgagcagg aaggcctggc cccctacctg agcagatggg agaagctgga 3601 caacttcatc aaccggcccg tgaaactgat catcggcgac aaagagatct tcggcatcag 3661 ccggggcatc gacaagcagg gggccctgct cctggaacag gacggcatca tcaagccctg 3721 gatgggcggc gagatcagcc tgcggagcgc cgagaagtga tgactcgagt ctagagggcc 3781 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 3841 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 3901 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3961 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 4021 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 4081 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 4141 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 4201 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 4261 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt 4321 cctattccga agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga 4381 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct 4441 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg 4501 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg 4561 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga 4621 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga 4681 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc 4741 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac 4801 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt 4861 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc 4921 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct 4981 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc 5041 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca 5101 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta 5161 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga 5221 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg 5281 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact 5341 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact 5401 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 5461 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 5521 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 5581 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 5641 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 5701 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 5761 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 5821 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 5881 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 5941 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 6001 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 6061 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 6121 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 6181 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 6241 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 6301 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 6361 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 6421 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 6481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 6541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 6601 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 7141 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 7441 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 7861 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 7921 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt 7981 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg 8041 cgttttgcgc tg //