LOCUS       sequence-212747-        7437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-03-22"; volume: "9(1):1188. doi"; pages: " 
            10.1038/s41467-018-03523-2"
COMMENT     Sequence Label: MAC_C_AOX
FEATURES             Location/Qualifiers
     source          1..7437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2150..2176
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2177..2524,2525..2527,2528..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
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     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(join(5119..5125,5126..5143,5144..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(join(6232..7023,7024..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
//
