LOCUS       sequence-212773-        6840 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 6840)
  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 6840)
  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_C_COX41
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        18..397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..601
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     552..572
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    603..621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    624..642
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     652..676
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    819..843
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1360..1384)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1431..1454
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1491..1514
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1545..1571
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1545..1571
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1572..1919,1920..1922,1923..2531)
                     /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="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(2575..2592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2581..2805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2938..2957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3089..3136
                     /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             3144..4166
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3144..4166
                     /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    4296..4417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4333..4352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4387..4406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4466..4482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4466..4482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4479..4501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4490..4506
                     /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(4514..4520,4521..4538,4539..4544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4559..4580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4697..4714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4868..5456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4948..4967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5627..6418,6419..6487))
                     /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     6250..6269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6488..6592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6667..6686)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
       61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      361 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      421 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      481 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      541 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      601 tctccctatc agtgatagag atctccctat cagtgataga gatcgtcgac gagctcgttt
      661 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      721 ccgggaccga tccagcctcc ggactctagc gtttaaactt aagcttggta ccgagctcgg
      781 atccactagt ccagtgtggt ggaattctgc agatatcaac aagtttgtac aaaaaagcag
      841 gctccaccat gttggctacc agggtattta gcctagttgg caagcgagca atttccacct
      901 ctgtgtgtgt acgagctcat gaaagtgttg tgaagagcga agacttttcg ctcccagctt
      961 atatggatcg gcgtgaccac cccttgccgg aggtggccca tgtcaagcac ctgtctgcca
     1021 gccagaaggc actgaaggag aaggagaagg cctcctggag cagcctctcc atggatgaga
     1081 aagtcgagtt gtatcgcatt aagttcaagg agagctttgc tgagatgaac aggggctcga
     1141 acgagtggaa gacggttgtg ggcggtgcca tgttcttcat cggtttcacc gcgctcgtta
     1201 tcatgtggca gaagcactat gtgtacggcc ccctcccgca aagctttgac aaagagtggg
     1261 tggccaagca gaccaagagg atgctggaca tgaaggtgaa ccccatccag ggcttagcct
     1321 ccaagtggga ctacgaaaag aacgagtgga agaagttgga cccagctttc ttgtacaaag
     1381 tggttgatat caaggccgcc gacatcacca gcctgtacaa gaaagtgggg tggagccatc
     1441 cccagttcga gaagggcgga ggatctggcg gcggaagcgg cggaggaagc tggtcccacc
     1501 ctcagtttga gaaaggcacc gagctgggca gcaccatggc cagctacccc tacgacgtgc
     1561 ccgactacgc caaggacaac accgtgcccc tgaagctgat cgccctgctg gccaacggcg
     1621 agttccactc tggcgagcag ctgggcgaga cactgggcat gagcagagcc gccatcaaca
     1681 agcacatcca gaccctgcgg gactggggcg tggacgtgtt cacagtgccc ggcaagggct
     1741 acagcctgcc cgagcccatc cagctgctga acgccaagca gatcctgggc cagctggatg
     1801 gcggatctgt ggccgtgctg cccgtgatcg acagcaccaa ccagtatctg ctcgaccgga
     1861 tcggcgagct gaagtccggc gacgcctgta tcgccgagta ccagcaggcc ggcagaggcg
     1921 gcaggggcag aaagtggttc agccccttcg gcgccaacct gtacctgagc atgttctgga
     1981 gactggaaca gggcccagcc gccgctattg gcctgagcct ggtcatcggc atcgtgatgg
     2041 ccgaggtgct gcggaagctg ggcgccgaca aagtgcgcgt gaagtggccc aacgacctgt
     2101 atctgcagga ccggaagctg gccggcatcc tggtggaact gaccggcaaa acaggcgacg
     2161 ccgcccagat cgtgatcggc gctggcatca acatggccat gcggcgggtg gaagagagcg
     2221 tggtcaacca gggctggatc accctgcagg aagccggcat caacctggac cggaacaccc
     2281 tggccgccat gctgatcaga gagctgagag ccgccctgga actgttcgag caggaaggcc
     2341 tggcccccta cctgagcaga tgggagaagc tggacaactt catcaaccgg cccgtgaaac
     2401 tgatcatcgg cgacaaagag atcttcggca tcagccgggg catcgacaag cagggggccc
     2461 tgctcctgga acaggacggc atcatcaagc cctggatggg cggcgagatc agcctgcgga
     2521 gcgccgagaa gtgatgactc gagtctagag ggcccgttta aacccgctga tcagcctcga
     2581 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2641 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2701 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2761 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2821 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2881 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2941 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3001 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3061 ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag
     3121 aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     3181 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     3241 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     3301 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     3361 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     3421 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     3481 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     3541 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     3601 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     3661 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     3721 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     3781 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     3841 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     3901 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     3961 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     4021 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     4081 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     4141 gcactcgtcc gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct
     4201 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4261 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4441 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4501 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4561 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4621 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4681 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4741 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4801 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4861 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4921 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4981 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5041 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5101 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5161 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5221 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5281 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5341 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5401 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5461 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5521 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5581 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5641 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5701 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5761 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5821 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5881 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5941 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6001 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6061 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6121 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6181 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6241 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6601 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6661 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6721 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6781 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
//
