LOCUS       sequence-214715-        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_TBG1
FEATURES             Location/Qualifiers
     source          1..7665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    594..612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    615..633
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     643..667
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(835..1185,1186..1188,1189..1797)
                     /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             1798..1824
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1855..1878
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1915..1938
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1951..1975
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3329..3353)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3391..3408)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3397..3621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3754..3773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3905..3952
                     /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             3960..4982
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3960..4982
                     /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    5112..5233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5149..5168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5203..5222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5282..5298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5282..5298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5295..5317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5306..5322
                     /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(5330..5336,5337..5354,5355..5360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5375..5396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5513..5530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5684..6272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5764..5783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6443..7234,7235..7303))
                     /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     7066..7085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7304..7408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7483..7502)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat
      601 cagtgataga gatctcccta tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg
      661 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
      721 atccagcctc cggactctag cgtttaaact taagcttggt accgagctcg gatccactag
      781 tccagtgtgg tggaattctg cagatatcca gcacagtggc ggccgctcga gaccatgaag
      841 gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca acggcgagtt ccactctggc
      901 gagcagctgg gcgagacact gggcatgagc agagccgcca tcaacaagca catccagacc
      961 ctgcgggact ggggcgtgga cgtgttcaca gtgcccggca agggctacag cctgcccgag
     1021 cccatccagc tgctgaacgc caagcagatc ctgggccagc tggatggcgg atctgtggcc
     1081 gtgctgcccg tgatcgacag caccaaccag tatctgctcg accggatcgg cgagctgaag
     1141 tccggcgacg cctgtatcgc cgagtaccag caggccggca gaggcggcag gggcagaaag
     1201 tggttcagcc ccttcggcgc caacctgtac ctgagcatgt tctggagact ggaacagggc
     1261 ccagccgccg ctattggcct gagcctggtc atcggcatcg tgatggccga ggtgctgcgg
     1321 aagctgggcg ccgacaaagt gcgcgtgaag tggcccaacg acctgtatct gcaggaccgg
     1381 aagctggccg gcatcctggt ggaactgacc ggcaaaacag gcgacgccgc ccagatcgtg
     1441 atcggcgctg gcatcaacat ggccatgcgg cgggtggaag agagcgtggt caaccagggc
     1501 tggatcaccc tgcaggaagc cggcatcaac ctggaccgga acaccctggc cgccatgctg
     1561 atcagagagc tgagagccgc cctggaactg ttcgagcagg aaggcctggc cccctacctg
     1621 agcagatggg agaagctgga caacttcatc aaccggcccg tgaaactgat catcggcgac
     1681 aaagagatct tcggcatcag ccggggcatc gacaagcagg gggccctgct cctggaacag
     1741 gacggcatca tcaagccctg gatgggcggc gagatcagcc tgcggagcgc cgagaagtac
     1801 ccatacgatg ttcctgacta tgccggtacc gagctcggat ccaccatggc tagctggagc
     1861 cacccgcagt tcgagaaagg tggaggttcc ggaggtggat cgggaggtgg atcgtggagc
     1921 cacccgcagt tcgaaaaagc ggccgatatc acaagtttgt acaaaaaagt tggcatgccg
     1981 agggaaatca tcaccctaca gttgggccag tgcggcaatc agattgggtt cgagttctgg
     2041 aaacagctgt gcgccgagca tggtatcagc cccgagggca tcgtggagga gttcgccacc
     2101 gagggcactg accgcaagga cgtctttttc taccaggcag acgatgagca ctacatcccc
     2161 cgggccgtgc tgctggactt ggaaccccgg gtgatccact ccatcctcaa ctccccctat
     2221 gccaagctct acaacccaga gaacatctac ctgtcggaac atggaggagg agctggcaac
     2281 aactgggcca gcggattctc ccagggagaa aagatccatg aggacatttt tgacatcata
     2341 gaccgggagg cagatggtag tgacagtcta gagggctttg tgctgtgtca ctccattgct
     2401 ggggggacag gctctggact gggttcctac ctcttagaac ggctgaatga caggtatcct
     2461 aagaagctgg tgcagacata ctcagtgttt cccaaccagg acgagatgag cgatgtggtg
     2521 gtccagcctt acaattcact cctcacactc aagaggctga cgcagaatgc agactgtgtg
     2581 gtggtgctgg acaacacagc cctgaaccgg attgccacag accgcctgca catccagaac
     2641 ccatccttct cccagatcaa ccagctggtg tctaccatca tgtcagccag caccaccacc
     2701 ctgcgctacc ctggctacat gaacaatgac ctcatcggcc tcatcgcctc gctcattccc
     2761 accccacggc tccacttcct catgaccggc tacacccctc tcactacgga ccagtcagtg
     2821 gccagcgtga ggaagaccac ggtcctggat gtcatgaggc ggctgctgca gcccaagaac
     2881 gtgatggtgt ccacaggccg agaccgccag accaaccact gctacatcgc catcctcaac
     2941 atcatccagg gagaggtgga ccccacccag gtccacaaga gcttgcagag gatccgggaa
     3001 cgcaagttgg ccaacttcat cccgtggggc cccgccagca tccaggtggc cctgtcgagg
     3061 aagtctccct acctgccctc ggcccaccgg gtcagcgggc tcatgatggc caaccacacc
     3121 agcatctcct cgctcttcga gagaacctgt cgccagtatg acaagctgcg taagcgggag
     3181 gccttcctgg agcagttccg caaggaggac atgttcaagg acaactttga tgagatggac
     3241 acatccaggg agattgtgca gcagctcatc gatgagtacc atgcggccac acggccagac
     3301 tacatctcct ggggcaccca ggagcagtgc ccaactttct tgtacaaagt ggtgacgtaa
     3361 gctaggggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3421 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3481 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3541 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3601 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3661 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3721 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3781 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3841 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3901 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
     3961 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     4021 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     4081 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     4141 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     4201 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     4261 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     4321 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     4381 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     4441 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     4501 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     4561 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     4621 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     4681 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     4741 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     4801 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     4861 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     4921 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     4981 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5041 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5101 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5161 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5221 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5281 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5341 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5401 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5461 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5521 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5581 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5641 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6121 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6721 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7261 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7321 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7381 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7441 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     7501 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7561 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     7621 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agata
//
