LOCUS Exported 7755 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION MAC-tagged gene expression. ACCESSION . VERSION . KEYWORDS MAC_C_TR61B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7755) 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 7755) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7755 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 23..402 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 403..606 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 557..577 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 608..626 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 629..647 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 657..681 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 824..848 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2280..2304) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2351..2374 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2411..2434 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2465..2491 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2492..3451 /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(3495..3512) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3501..3725 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3858..3877) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 4009..4056 /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 4064..5086 /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 5216..5337 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5253..5272) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5307..5326 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5386..5402) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5386..5402) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5399..5421) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5410..5426 /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(5434..5464) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5479..5500 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5617..5634) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5788..6376) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5868..5887) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6547..7407) /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 7170..7189 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7408..7512) /gene="bla" /label=AmpR promoter primer_bind complement(7587..7606) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt 61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc 301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct 361 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag 421 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt 481 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac 541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc 601 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct 661 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga 721 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tggtaccgag 781 ctcggatcca ctagtccagt gtggtggaat tctgcagata tcaacaagtt tgtacaaaaa 841 agttggcatg ctaatggcat ggtgccgcgg tcctgtcttg ctgtgcctgc ggcaggggct 901 cggaaccaat tcattcctgc acggcctggg gcaggagccc ttcgagggag ctcggtcact 961 gtgttgcagg tcctcgccta gagacctgcg agatggagaa agagagcacg aggcggcaca 1021 aaggaaagcc ccaggagcag agtcttgccc atctctccct ctgagcatct cggacattgg 1081 gactggatgt ctttcgtcac tggaaaacct cagactgccg acgctgcggg aagagtcatc 1141 acctcgagag ctcgaggact cgagcggaga ccagggccgg tgcggtccca cacaccaggg 1201 atccgaggat ccttcgatgc tctcgcaggc ccagtccgct accgaggtcg aagagcgtca 1261 cgtctcccct tcttgttcaa cttccagaga gagacccttt caggctgggg agctgatttt 1321 agctgagact ggggagggag aaacaaaatt taagaaatta tttaggttga acaacttcgg 1381 actcttaaat agtaactggg gggcagtccc gttcggcaag atcgtgggga agttccccgg 1441 ccagatactg aggagttcct tcggtaagca gtacatgctg aggaggccag ccttggaaga 1501 ctatgtagta ttgatgaaaa gagggactgc cataacattc ccaaaggata ttaatatgat 1561 tctctcaatg atggatatca acccaggtga tactgttttg gaagctggct caggctctgg 1621 tggaatgagc ttatttttat ccaaagcagt tggatcacaa ggacgagtca taagttttga 1681 ggtacgaaaa gaccaccatg atctggctaa gaagaattac aaacactggc gtgattcatg 1741 gaaattaagt catgtagaag agtggccaga caatgtggat tttattcata aggacatttc 1801 aggagcaacc gaagacataa aatctttaac atttgacgca gtagctttgg atatgttaaa 1861 tcctcatgtt actttgcctg ttttttaccc acatcttaag catggtggtg tatgtgctgt 1921 atatgtagta aacatcacac aggttattga acttttagat ggaattcgca cctgtgaact 1981 tgctctttca tgtgaaaaga taagcgaggt cattgtcaga gattggttgg tttgccttgc 2041 aaaacagaaa aatggaattt tagctcaaaa agtagaatct aaaatcaaca cagatgtaca 2101 actagattct caagagaaaa ttggagttaa aggtgagctg tttcaagagg atgaccatga 2161 agaatcgcat tctgattttc catatggatc atttccctat gttgctagac cagtacactg 2221 gcaacctggt catacagctt ttcttgtcaa gttgaggaag gtcaaaccac aacttaactg 2281 cccaactttc ttgtacaaag tggttgatat caaggccgcc gacatcacca gcctgtacaa 2341 gaaagtgggg tggagccatc cccagttcga gaagggcgga ggatctggcg gcggaagcgg 2401 cggaggaagc tggtcccacc ctcagtttga gaaaggcacc gagctgggca gcaccatggc 2461 cagctacccc tacgacgtgc ccgactacgc caaggacaac accgtgcccc tgaagctgat 2521 cgccctgctg gccaacggcg agttccactc tggcgagcag ctgggcgaga cactgggcat 2581 gagcagagcc gccatcaaca agcacatcca gaccctgcgg gactggggcg tggacgtgtt 2641 cacagtgccc ggcaagggct acagcctgcc cgagcccatc cagctgctga acgccaagca 2701 gatcctgggc cagctggatg gcggatctgt ggccgtgctg cccgtgatcg acagcaccaa 2761 ccagtatctg ctcgaccgga tcggcgagct gaagtccggc gacgcctgta tcgccgagta 2821 ccagcaggcc ggcagaggcg gcaggggcag aaagtggttc agccccttcg gcgccaacct 2881 gtacctgagc atgttctgga gactggaaca gggcccagcc gccgctattg gcctgagcct 2941 ggtcatcggc atcgtgatgg ccgaggtgct gcggaagctg ggcgccgaca aagtgcgcgt 3001 gaagtggccc aacgacctgt atctgcagga ccggaagctg gccggcatcc tggtggaact 3061 gaccggcaaa acaggcgacg ccgcccagat cgtgatcggc gctggcatca acatggccat 3121 gcggcgggtg gaagagagcg tggtcaacca gggctggatc accctgcagg aagccggcat 3181 caacctggac cggaacaccc tggccgccat gctgatcaga gagctgagag ccgccctgga 3241 actgttcgag caggaaggcc tggcccccta cctgagcaga tgggagaagc tggacaactt 3301 catcaaccgg cccgtgaaac tgatcatcgg cgacaaagag atcttcggca tcagccgggg 3361 catcgacaag cagggggccc tgctcctgga acaggacggc atcatcaagc cctggatggg 3421 cggcgagatc agcctgcgga gcgccgagaa gtgatgactc gagtctagag ggcccgttta 3481 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc 3541 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 3601 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 3661 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 3721 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg 3781 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 3841 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 3901 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 3961 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt 4021 ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac 4081 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca 4141 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt 4201 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt 4261 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct 4321 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga 4381 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct 4441 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg 4501 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga 4561 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga 4621 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga 4681 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata 4741 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg 4801 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct 4861 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc 4921 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac 4981 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga 5041 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga 5101 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 5161 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt 5221 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 5281 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 5341 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc 5401 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 5461 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 5521 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 5581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 5641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 5701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 5761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 5821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 5881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 6241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 6421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 6481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 6541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 6601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 6661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 6721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 6781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 6841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 7321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 7381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 7441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 7501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct 7561 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 7621 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct 7681 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt 7741 gcgctgcttc gcgat //