LOCUS       sequence-212857-        7356 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 7356)
  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 7356)
  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_CDK7
FEATURES             Location/Qualifiers
     source          1..7356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        21..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    606..624
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    627..645
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     655..679
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(847..1197,1198..1200,1201..1809)
                     /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             1810..1836
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1867..1890
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1927..1950
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1963..1987
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3032..3056)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3094..3111)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3100..3324
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3457..3476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3608..3655
                     /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             3663..4685
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3663..4685
                     /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    4815..4936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4852..4871)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4906..4925
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4985..5001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4985..5001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4998..5020)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5009..5025
                     /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(5033..5039,5040..5057,5058..5063))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5078..5099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5216..5233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5387..5975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5467..5486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6146..6937,6938..7006))
                     /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     6769..6788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7007..7111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7186..7205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctctccct atcagtgata gagatctccc tatcagtgat agagatcgtc gacgagctcg
      661 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
      721 acaccgggac cgatccagcc tccggactct agcgtttaaa cttaagcttg gtaccgagct
      781 cggatccact agtccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgctc
      841 gagaccatga aggacaacac cgtgcccctg aagctgatcg ccctgctggc caacggcgag
      901 ttccactctg gcgagcagct gggcgagaca ctgggcatga gcagagccgc catcaacaag
      961 cacatccaga ccctgcggga ctggggcgtg gacgtgttca cagtgcccgg caagggctac
     1021 agcctgcccg agcccatcca gctgctgaac gccaagcaga tcctgggcca gctggatggc
     1081 ggatctgtgg ccgtgctgcc cgtgatcgac agcaccaacc agtatctgct cgaccggatc
     1141 ggcgagctga agtccggcga cgcctgtatc gccgagtacc agcaggccgg cagaggcggc
     1201 aggggcagaa agtggttcag ccccttcggc gccaacctgt acctgagcat gttctggaga
     1261 ctggaacagg gcccagccgc cgctattggc ctgagcctgg tcatcggcat cgtgatggcc
     1321 gaggtgctgc ggaagctggg cgccgacaaa gtgcgcgtga agtggcccaa cgacctgtat
     1381 ctgcaggacc ggaagctggc cggcatcctg gtggaactga ccggcaaaac aggcgacgcc
     1441 gcccagatcg tgatcggcgc tggcatcaac atggccatgc ggcgggtgga agagagcgtg
     1501 gtcaaccagg gctggatcac cctgcaggaa gccggcatca acctggaccg gaacaccctg
     1561 gccgccatgc tgatcagaga gctgagagcc gccctggaac tgttcgagca ggaaggcctg
     1621 gccccctacc tgagcagatg ggagaagctg gacaacttca tcaaccggcc cgtgaaactg
     1681 atcatcggcg acaaagagat cttcggcatc agccggggca tcgacaagca gggggccctg
     1741 ctcctggaac aggacggcat catcaagccc tggatgggcg gcgagatcag cctgcggagc
     1801 gccgagaagt acccatacga tgttcctgac tatgccggta ccgagctcgg atccaccatg
     1861 gctagctgga gccacccgca gttcgagaaa ggtggaggtt ccggaggtgg atcgggaggt
     1921 ggatcgtgga gccacccgca gttcgaaaaa gcggccgata tcacaagttt gtacaaaaaa
     1981 gcaggctcca ccatggctct ggacgtgaag tctcgggcaa agcgttatga gaagctggac
     2041 ttccttgggg agggacagtt tgccaccgtt tacaaggcca gagataagaa taccaaccaa
     2101 attgtcgcca ttaagaaaat caaacttgga catagatcag aagctaaaga tggtataaat
     2161 agaaccgcct taagagagat aaaattatta caggagctaa gtcatccaaa tataattggt
     2221 ctccttgatg cttttggaca taaatctaat attagccttg tctttgattt tatggaaact
     2281 gatctagagg ttataataaa ggataatagt cttgtgctga caccatcaca catcaaagcc
     2341 tacatgttga tgactcttca aggattagaa tatttacatc aacattggat cctacatagg
     2401 gatctgaaac caaacaactt gttgctagat gaaaatggag ttctaaaact ggcagatttt
     2461 ggcctggcca aatcttttgg gagccccaat agagcttata cacatcaggt tgtaaccagg
     2521 tggtatcggg cccccgagtt actatttgga gctaggatgt atggtgtagg tgtggacatg
     2581 tgggctgttg gctgtatatt agcagagtta cttctaaggg ttcctttttt gccaggagat
     2641 tcagaccttg atcagctaac aagaatattt gaaactttgg gcacaccaac tgaggaacag
     2701 tggccggaca tgtgtagtct tccagattat gtgacattta agagtttccc tggaatacct
     2761 ttgcatcaca tcttcagtgc agcaggagac gacttactag atctcataca aggcttattc
     2821 ttatttaatc catgtgctcg aattacggcc acacaggcac tgaaaatgaa gtatttcagt
     2881 aatcggccag ggccaacacc tggatgtcag ctgccaagac caaactgtcc agtggaaacc
     2941 ttaaaggagc aatcaaatcc agctttggca ataaaaagga aaagaacaga ggccttagaa
     3001 caaggaggat tgcccaagaa actaattttt gacccagctt tcttgtacaa agtggtgacg
     3061 taagctaggg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3121 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3181 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3241 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3301 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3361 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3421 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3481 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3541 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3601 gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg
     3661 ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     3721 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     3781 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     3841 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     3901 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     3961 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     4021 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     4081 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     4141 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     4201 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     4261 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     4321 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     4381 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     4441 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     4501 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     4561 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     4621 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     4681 aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4741 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4801 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4861 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4921 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4981 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5041 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5101 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5161 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5221 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5281 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5341 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5401 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5461 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5521 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5581 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5641 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5701 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5761 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5821 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5881 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5941 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6001 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6061 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6121 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6181 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6241 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6301 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6361 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6421 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6481 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6541 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6601 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6661 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6721 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6781 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6841 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6901 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6961 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7021 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7081 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7141 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     7201 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     7261 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     7321 gcttagggtt aggcgttttg cgctgcttcg cgatgt
//
