LOCUS       sequence-212878-        7059 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 7059)
  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 7059)
  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_N_RS6
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    599..617
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    620..638
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     648..672
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(840..1190,1191..1193,1194..1802)
                     /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="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1803..1829
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1860..1883
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1920..1943
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1956..1980
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2728..2752)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2790..2807)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2796..3020
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3153..3172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3304..3351
                     /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             3359..4381
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3359..4381
                     /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    4511..4632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4548..4567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4602..4621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4681..4697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4681..4697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4694..4716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4705..4721
                     /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(4729..4735,4736..4753,4754..4759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4774..4795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4912..4929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5083..5671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5163..5182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5842..6633,6634..6702))
                     /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     6465..6484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6703..6807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6882..6901)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      601 cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg
      661 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      721 gaccgatcca gcctccggac tctagcgttt aaacttaagc ttggtaccga gctcggatcc
      781 actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagacca
      841 tgaaggacaa caccgtgccc ctgaagctga tcgccctgct ggccaacggc gagttccact
      901 ctggcgagca gctgggcgag acactgggca tgagcagagc cgccatcaac aagcacatcc
      961 agaccctgcg ggactggggc gtggacgtgt tcacagtgcc cggcaagggc tacagcctgc
     1021 ccgagcccat ccagctgctg aacgccaagc agatcctggg ccagctggat ggcggatctg
     1081 tggccgtgct gcccgtgatc gacagcacca accagtatct gctcgaccgg atcggcgagc
     1141 tgaagtccgg cgacgcctgt atcgccgagt accagcaggc cggcagaggc ggcaggggca
     1201 gaaagtggtt cagccccttc ggcgccaacc tgtacctgag catgttctgg agactggaac
     1261 agggcccagc cgccgctatt ggcctgagcc tggtcatcgg catcgtgatg gccgaggtgc
     1321 tgcggaagct gggcgccgac aaagtgcgcg tgaagtggcc caacgacctg tatctgcagg
     1381 accggaagct ggccggcatc ctggtggaac tgaccggcaa aacaggcgac gccgcccaga
     1441 tcgtgatcgg cgctggcatc aacatggcca tgcggcgggt ggaagagagc gtggtcaacc
     1501 agggctggat caccctgcag gaagccggca tcaacctgga ccggaacacc ctggccgcca
     1561 tgctgatcag agagctgaga gccgccctgg aactgttcga gcaggaaggc ctggccccct
     1621 acctgagcag atgggagaag ctggacaact tcatcaaccg gcccgtgaaa ctgatcatcg
     1681 gcgacaaaga gatcttcggc atcagccggg gcatcgacaa gcagggggcc ctgctcctgg
     1741 aacaggacgg catcatcaag ccctggatgg gcggcgagat cagcctgcgg agcgccgaga
     1801 agtacccata cgatgttcct gactatgccg gtaccgagct cggatccacc atggctagct
     1861 ggagccaccc gcagttcgag aaaggtggag gttccggagg tggatcggga ggtggatcgt
     1921 ggagccaccc gcagttcgaa aaagcggccg atatcacaag tttgtacaaa aaagttggca
     1981 tgaagctgaa catctccttc ccagccactg gctgccagaa actcattgaa gtggacgatg
     2041 aacgcaaact tcgtactttc tatgagaagc gtatggccac agaagttgct gctgacgctc
     2101 tgggtgaaga atggaagggt tatgtggtcc gaatcagtgg tgggaacgac aaacaaggtt
     2161 tccccatgaa gcagggtgtc ttgacccatg gccgtgtccg cctgctactg agtaaggggc
     2221 attcctgtta cagaccaagg agaactggag aaagaaagag aaaatcagtt cgtggttgca
     2281 ttgtggatgc aaatctgagc gttctcaact tggttattgt aaaaaaagga gagaaggata
     2341 ttcctggact gactgatact acagtgcctc gccgcctggg ccccaaaaga gctagcagaa
     2401 tccgcaaact tttcaatctc tctaaagaag atgatgtccg ccagtatgtt gtaagaaagc
     2461 ccttaaataa agaaggtaag aaacctagga ccaaagcacc caagattcag cgtcttgtta
     2521 ctccacgtgt cctgcagcac aaacggcggc gtattgctct gaagaagcag cgtaccaaga
     2581 aaaataaaga agaggctgca gaatatgcta aacttttggc caagagaatg aaggaggcta
     2641 aggagaagcg ccaggaacaa attgcgaaga gacgcagact ttcctctctg cgagcttcta
     2701 cttctaagtc tgaatccagt cagaaatacc caactttctt gtacaaagtg gtgacgtaag
     2761 ctaggggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2821 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2881 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2941 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3001 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3061 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3121 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3181 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3241 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     3301 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     3361 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3421 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3481 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3541 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3601 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3661 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3721 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3781 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3841 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3901 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     3961 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4021 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4081 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     4141 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4201 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4261 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4321 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4381 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4441 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4501 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4561 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4621 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4681 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4741 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4801 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     6841 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     6901 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     6961 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7021 agggttaggc gttttgcgct gcttcgcgat gtacgggcc
//
