LOCUS Exported 7437 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION MAC-tagged gene expression. ACCESSION . VERSION . KEYWORDS MAC_C_AOX SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7437) 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 7437) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7437 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 26..405 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 406..609 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 560..580 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 611..629 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 632..650 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 660..684 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 827..851 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(1965..1989) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2036..2059 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2096..2119 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2150..2176 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2177..3136 /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(3180..3197) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3186..3410 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3543..3562) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3694..3741 /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 3749..4771 /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 4901..5022 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4938..4957) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4992..5011 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5071..5087) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5071..5087) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5084..5106) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5095..5111 /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(5119..5149) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5164..5185 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5302..5319) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5473..6061) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5553..5572) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6232..7092) /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 6855..6874 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7093..7197) /gene="bla" /label=AmpR promoter primer_bind complement(7272..7291) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca 61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta 121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat 181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg 241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac 301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt 361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg 421 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc 481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt 541 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata 601 agcagagctc tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga 661 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 721 agaagacacc gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc 781 gagctcggat ccactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa 841 aaaagcaggc tccaccatgc tgtctaccgg cagcaagacc ttcctgttcc ggccattcct 901 gggctcctgc cacgccctgc agagcggcaa gctgccctgc agcaacctgc acaccacccc 961 taccaagatc accgtgaaga gatacctcgt gggatacagc tggtccaccc agccccacag 1021 cagactgctc cacagctgcc agcagctgaa gatcgacgac aagaacaaga gcgagcactt 1081 caagatcgag acaaacgaca gcaccgacga gcccaacatc gaggtggaaa acttccctca 1141 cttcagagag gccaagaagg ccaaagagac acagaagggc agcagtctgg ccgaggccga 1201 ggaacacccc gacgtggaag agggcagagc catgcaggac ggcggctaca gactgcccca 1261 ccccatctgg cacaagcagg aactcgaaag cgtccggatc agccaccggc ctcccgtggg 1321 caaggtggac aagctggcct actacagcgt gcagctgctg cggaccggct tcgacgtgtt 1381 cagcggctac accctgggca cctacaccgg cagactggac gagaagcagt gggtgaaacg 1441 gatcatcttc ctggaaacaa tcgccggcgt gccaggcatg gtcggagcta tggtccgcca 1501 cctggtgtcc ctgcggagac tgaagcggga ccacggctgg atccacaccc tgctggaaga 1561 ggccgagaac gagcggatgc acctgatgac cgccatgcgg atcgccaacc ccggcatcat 1621 catgcggacc agcatcgtgg tggcccaggg catcttcgtg tccggcttca gcctggccta 1681 cctgatcagc cccagattct gccacagatt cgtgggctac ctggaagagg aagccgtgaa 1741 aacctacacc cactgcctgg aagaactgga cagcggcaac ctgaagatgt ggtgccggat 1801 gaaggcccct gagatcgccg tggaatactg gaagctgccc gacgacgcca tgatgcggga 1861 cgtgatcctg gccatcagag ccgacgaggc ccaccacaga agcgtgaacc acgacctggg 1921 cagccggaag cccgacgagc agaaccctta ccctcccggc caggacccag ctttcttgta 1981 caaagtggtt gatatcaagg ccgccgacat caccagcctg tacaagaaag tggggtggag 2041 ccatccccag ttcgagaagg gcggaggatc tggcggcgga agcggcggag gaagctggtc 2101 ccaccctcag tttgagaaag gcaccgagct gggcagcacc atggccagct acccctacga 2161 cgtgcccgac tacgccaagg acaacaccgt gcccctgaag ctgatcgccc tgctggccaa 2221 cggcgagttc cactctggcg agcagctggg cgagacactg ggcatgagca gagccgccat 2281 caacaagcac atccagaccc tgcgggactg gggcgtggac gtgttcacag tgcccggcaa 2341 gggctacagc ctgcccgagc ccatccagct gctgaacgcc aagcagatcc tgggccagct 2401 ggatggcgga tctgtggccg tgctgcccgt gatcgacagc accaaccagt atctgctcga 2461 ccggatcggc gagctgaagt ccggcgacgc ctgtatcgcc gagtaccagc aggccggcag 2521 aggcggcagg ggcagaaagt ggttcagccc cttcggcgcc aacctgtacc tgagcatgtt 2581 ctggagactg gaacagggcc cagccgccgc tattggcctg agcctggtca tcggcatcgt 2641 gatggccgag gtgctgcgga agctgggcgc cgacaaagtg cgcgtgaagt ggcccaacga 2701 cctgtatctg caggaccgga agctggccgg catcctggtg gaactgaccg gcaaaacagg 2761 cgacgccgcc cagatcgtga tcggcgctgg catcaacatg gccatgcggc gggtggaaga 2821 gagcgtggtc aaccagggct ggatcaccct gcaggaagcc ggcatcaacc tggaccggaa 2881 caccctggcc gccatgctga tcagagagct gagagccgcc ctggaactgt tcgagcagga 2941 aggcctggcc ccctacctga gcagatggga gaagctggac aacttcatca accggcccgt 3001 gaaactgatc atcggcgaca aagagatctt cggcatcagc cggggcatcg acaagcaggg 3061 ggccctgctc ctggaacagg acggcatcat caagccctgg atgggcggcg agatcagcct 3121 gcggagcgcc gagaagtgat gactcgagtc tagagggccc gtttaaaccc gctgatcagc 3181 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3241 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3301 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3361 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3421 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3481 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3541 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3601 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 3661 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc 3721 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga 3781 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga 3841 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag 3901 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct 3961 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc 4021 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct 4081 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg 4141 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg 4201 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc 4261 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg 4321 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac 4381 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat 4441 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag 4501 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga 4561 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg 4621 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag 4681 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg 4741 ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc 4801 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 4861 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 4921 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4981 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 5041 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5101 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5161 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5221 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 5281 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 5341 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 5401 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5461 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5521 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5581 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5641 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5701 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5761 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5821 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5881 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5941 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6001 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6061 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 6121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 6181 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 6241 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 6301 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 6361 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 6421 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 6481 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6541 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 6601 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 6661 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 6721 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 6781 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6841 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 6901 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 6961 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7021 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7081 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 7141 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 7201 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg 7261 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct 7321 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc 7381 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgc //