LOCUS       sequence-212881-        7665 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 7665)
  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 7665)
  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_TBA1A
FEATURES             Location/Qualifiers
     source          1..7665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        405..608
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     559..579
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    610..628
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    631..649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     659..683
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(851..1201,1202..1204,1205..1813)
                     /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             1814..1840
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1871..1894
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1931..1954
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1967..1991
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3345..3369)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3407..3424)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3413..3637
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3770..3789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3921..3968
                     /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             3976..4998
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3976..4998
                     /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    5128..5249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5165..5184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5219..5238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5298..5314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5298..5314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5311..5333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5322..5338
                     /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(5346..5352,5353..5370,5371..5376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5391..5412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5529..5546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5700..6288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5780..5799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6459..7250,7251..7319))
                     /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     7082..7101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7320..7424)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7499..7518)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
       61 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      421 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      481 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      541 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      601 gcagagctct ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag
      661 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
      721 gaagacaccg ggaccgatcc agcctccgga ctctagcgtt taaacttaag cttggtaccg
      781 agctcggatc cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc
      841 gctcgagacc atgaaggaca acaccgtgcc cctgaagctg atcgccctgc tggccaacgg
      901 cgagttccac tctggcgagc agctgggcga gacactgggc atgagcagag ccgccatcaa
      961 caagcacatc cagaccctgc gggactgggg cgtggacgtg ttcacagtgc ccggcaaggg
     1021 ctacagcctg cccgagccca tccagctgct gaacgccaag cagatcctgg gccagctgga
     1081 tggcggatct gtggccgtgc tgcccgtgat cgacagcacc aaccagtatc tgctcgaccg
     1141 gatcggcgag ctgaagtccg gcgacgcctg tatcgccgag taccagcagg ccggcagagg
     1201 cggcaggggc agaaagtggt tcagcccctt cggcgccaac ctgtacctga gcatgttctg
     1261 gagactggaa cagggcccag ccgccgctat tggcctgagc ctggtcatcg gcatcgtgat
     1321 ggccgaggtg ctgcggaagc tgggcgccga caaagtgcgc gtgaagtggc ccaacgacct
     1381 gtatctgcag gaccggaagc tggccggcat cctggtggaa ctgaccggca aaacaggcga
     1441 cgccgcccag atcgtgatcg gcgctggcat caacatggcc atgcggcggg tggaagagag
     1501 cgtggtcaac cagggctgga tcaccctgca ggaagccggc atcaacctgg accggaacac
     1561 cctggccgcc atgctgatca gagagctgag agccgccctg gaactgttcg agcaggaagg
     1621 cctggccccc tacctgagca gatgggagaa gctggacaac ttcatcaacc ggcccgtgaa
     1681 actgatcatc ggcgacaaag agatcttcgg catcagccgg ggcatcgaca agcagggggc
     1741 cctgctcctg gaacaggacg gcatcatcaa gccctggatg ggcggcgaga tcagcctgcg
     1801 gagcgccgag aagtacccat acgatgttcc tgactatgcc ggtaccgagc tcggatccac
     1861 catggctagc tggagccacc cgcagttcga gaaaggtgga ggttccggag gtggatcggg
     1921 aggtggatcg tggagccacc cgcagttcga aaaagcggcc gatatcacaa gtttgtacaa
     1981 aaaagttggc atgcgtgagt gcatctccat ccacgttggc caggctggtg tccagattgg
     2041 caatgcctgc tgggagctct actgcctgga acacggcatc cagcccgatg gccagatgcc
     2101 aagtgacaag accattgggg gaggagatga ttccttcaac accttcttca gtgagacggg
     2161 ggctggcaag catgtgcccc gggcagtgtt tgtagacttg gaacccacag tcattgatga
     2221 agttcgcact ggcacctacc gccagctctt ccaccctgag caacttatca caggcaaaga
     2281 agatgctgcc aataactatg cccgagggca ctacaccatt ggcaaggaga tcattgacct
     2341 cgtgttggac cgaattcgca agctggccga ccagtgcacg ggtctccagg gcttcttggt
     2401 tttccacagc tttggtgggg gaactggttc tgggttcacc tcgctgctca tggaacgtct
     2461 ctcagttgat tatggcaaga agtccaagct ggagttctct atttacccgg cgccccaggt
     2521 ttccacagct gtagttgagc cctacaactc catcctcacc acccacacca ccctggagca
     2581 ctctgattgt gccttcatgg tagacaatga ggccatctat gacatctgtc gtagaaacct
     2641 cgatattgag cgtccaacct atactaacct gaataggtta ataggtcaaa ttgtgtcctc
     2701 catcactgct tccctgagat ttgatggagc cctgaatgtt gacctgacag aattccagac
     2761 caacctggtg ccctatcccc gcatccactt ccctctggcc acatatgccc ctgtcatctc
     2821 tgctgagaaa gcctaccatg aacagctttc tgtagcagag atcaccaatg cttgctttga
     2881 gccagccaac cagatggtga aatgtgaccc tcgccatggt aaatacatgg cttgctgcct
     2941 gttgtaccgt ggtgacgtgg ttcccaaaga tgtcaatgct gccattgcca ccatcaagac
     3001 caagcgtacc atccagtttg tggattggtg ccccactggc ttcaaggttg gcatcaacta
     3061 ccagcctccc actgtggtgc ctggtggaga cctggccaag gtacagagag ctgtgtgcat
     3121 gctgagcaac accacagcca ttgctgaggc ctgggctcgc ctggaccaca agtttgacct
     3181 gatgtatgcc aaacgtgcct ttgttcactg gtacgttggg gaggggatgg aggaaggtga
     3241 gttttcagag gcccgtgagg acatggctgc ccttgagaag gattatgagg aggttggtgt
     3301 ggattctgtt gaaggagagg gtgaggaaga aggagaggaa tactacccaa ctttcttgta
     3361 caaagtggtg acgtaagcta ggggcccgtt taaacccgct gatcagcctc gactgtgcct
     3421 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3481 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3541 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3601 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3661 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3721 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3781 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3841 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3901 ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct agaaagtata
     3961 ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     4021 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     4081 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     4141 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     4201 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     4261 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     4321 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     4381 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     4441 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     4501 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     4561 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     4621 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     4681 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     4741 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     4801 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     4861 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     4921 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     4981 ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc cttctatgaa
     5041 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5101 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5161 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5221 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5281 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5341 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5401 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5461 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5521 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5581 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5641 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5701 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5761 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5821 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5881 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5941 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6001 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6061 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6121 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6181 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6241 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6301 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6361 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6421 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6481 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6541 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6601 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6661 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6721 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6781 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6841 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6901 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6961 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7021 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7081 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7141 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7201 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7261 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7321 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7381 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7441 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     7501 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7561 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     7621 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcg
//
