LOCUS       sequence-212858-        7706 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 7706)
  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 7706)
  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_CDK8
FEATURES             Location/Qualifiers
     source          1..7706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     550..570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    601..619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    622..640
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     650..674
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(842..1192,1193..1195,1196..1804)
                     /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             1805..1831
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1862..1885
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1922..1945
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1958..1982
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3377..3401)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3439..3456)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3445..3669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3802..3821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3953..4000
                     /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             4008..5030
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4008..5030
                     /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    5160..5281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5197..5216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5251..5270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5330..5346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5330..5346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5343..5365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5354..5370
                     /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(5378..5384,5385..5402,5403..5408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5423..5444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5561..5578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5732..6320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5812..5831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6491..7282,7283..7351))
                     /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     7114..7133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7352..7456)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7531..7550)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      601 tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga gctcgtttag
      661 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      721 gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc gagctcggat
      781 ccactagtcc agtgtggtgg aattctgcag atatccagca cagtggcggc cgctcgagac
      841 catgaaggac aacaccgtgc ccctgaagct gatcgccctg ctggccaacg gcgagttcca
      901 ctctggcgag cagctgggcg agacactggg catgagcaga gccgccatca acaagcacat
      961 ccagaccctg cgggactggg gcgtggacgt gttcacagtg cccggcaagg gctacagcct
     1021 gcccgagccc atccagctgc tgaacgccaa gcagatcctg ggccagctgg atggcggatc
     1081 tgtggccgtg ctgcccgtga tcgacagcac caaccagtat ctgctcgacc ggatcggcga
     1141 gctgaagtcc ggcgacgcct gtatcgccga gtaccagcag gccggcagag gcggcagggg
     1201 cagaaagtgg ttcagcccct tcggcgccaa cctgtacctg agcatgttct ggagactgga
     1261 acagggccca gccgccgcta ttggcctgag cctggtcatc ggcatcgtga tggccgaggt
     1321 gctgcggaag ctgggcgccg acaaagtgcg cgtgaagtgg cccaacgacc tgtatctgca
     1381 ggaccggaag ctggccggca tcctggtgga actgaccggc aaaacaggcg acgccgccca
     1441 gatcgtgatc ggcgctggca tcaacatggc catgcggcgg gtggaagaga gcgtggtcaa
     1501 ccagggctgg atcaccctgc aggaagccgg catcaacctg gaccggaaca ccctggccgc
     1561 catgctgatc agagagctga gagccgccct ggaactgttc gagcaggaag gcctggcccc
     1621 ctacctgagc agatgggaga agctggacaa cttcatcaac cggcccgtga aactgatcat
     1681 cggcgacaaa gagatcttcg gcatcagccg gggcatcgac aagcaggggg ccctgctcct
     1741 ggaacaggac ggcatcatca agccctggat gggcggcgag atcagcctgc ggagcgccga
     1801 gaagtaccca tacgatgttc ctgactatgc cggtaccgag ctcggatcca ccatggctag
     1861 ctggagccac ccgcagttcg agaaaggtgg aggttccgga ggtggatcgg gaggtggatc
     1921 gtggagccac ccgcagttcg aaaaagcggc cgatatcaca agtttgtaca aaaaagcagg
     1981 ctccaccatg gactatgact ttaaagtaaa gctgagcagc gagcgggagc gggtcgagga
     2041 cctgtttgaa tacgagggct gcaaagttgg ccgaggcact tatggtcacg tctacaaagc
     2101 caagaggaaa gatgggaagg atgataaaga ctatgcttta aaacaaatag aaggaactgg
     2161 gatctctatg tcggcatgta gagaaatagc attacttcga gagcttaagc atccaaacgt
     2221 catttctctt caaaaggtgt ttctgtctca tgctgatagg aaggtgtggc ttctgtttga
     2281 ctatgctgaa catgacctct ggcatataat caagtttcac agagcttcta aagcaaacaa
     2341 gaagccagtt cagttacctc ggggaatggt gaagtcacta ttatatcaga tcctagatgg
     2401 tattcactac ctgcatgcta actgggtgtt gcacagagat ttgaaacctg ctaatatttt
     2461 agttatgggt gaaggtcctg agcgaggaag agtaaaaatt gctgacatgg gctttgcccg
     2521 attatttaat tcacctttga agcctttagc agatttggat ccagtggttg ttacattctg
     2581 gtaccgagcc cctgaactac ttcttggagc aaggcattat accaaagcta ttgatatttg
     2641 ggctataggg tgtatatttg cagaactact aacgtcagaa ccaatatttc actgtcgaca
     2701 agaggacatc aaaactagta atccttatca ccatgaccag ctggacagaa tattcaatgt
     2761 aatgggattt cctgcagata aagattggga agatataaaa aagatgcctg aacattcaac
     2821 attaatgaaa gatttcagaa gaaatacgta taccaactgc agccttatca agtatatgga
     2881 aaaacataaa gttaaaccag atagtaaagc attccacttg cttcagaagc tgcttaccat
     2941 ggacccaata aagcgaatta cctcagaaca ggctatgcag gacccctatt tcttagaaga
     3001 cccacttcct acatcagacg tttttgccgg ttgtcaaatc ccttacccaa aacgagaatt
     3061 tttaacggaa gaagaacctg atgacaaagg agacaaaaac cagcagcagc agcagggcaa
     3121 taaccacact aatggaactg gccacccagg gaatcaagac agcagtcaca cacagggacc
     3181 cccgttgaag aaagtgagag ttgttcctcc taccactacc tcaggtggac ttatcatgac
     3241 ctcagactat cagcgttcca atccacatgc tgcctatccc aaccctggac caagcacatc
     3301 acagccgcag agcagcatgg gatactcagc tacctcccag cagcctccac agtactcaca
     3361 tcagacacac ggtacgaccc agctttcttg tacaaagtgg tgacgtaagc taggggcccg
     3421 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3481 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3541 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3601 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3661 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3721 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3781 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3841 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3901 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc
     3961 tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac
     4021 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     4081 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     4141 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     4201 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     4261 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     4321 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     4381 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     4441 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     4501 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     4561 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     4621 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     4681 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     4741 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     4801 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     4861 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     4921 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     4981 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac
     5041 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5101 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5161 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5221 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5281 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5341 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5461 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5521 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6361 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7501 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7561 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7621 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     7681 ttttgcgctg cttcgcgatg tacggg
//
