LOCUS       sequence-212836-        7296 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 7296)
  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 7296)
  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_SFXN1
FEATURES             Location/Qualifiers
     source          1..7296
                     /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(1816..1840)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1887..1910
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1947..1970
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2001..2027
                     /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             2001..2027
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2028..2375,2376..2378,2379..2987)
                     /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(3031..3048)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3037..3261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3394..3413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3545..3592
                     /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             3600..4622
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3600..4622
                     /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    4752..4873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4789..4808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4843..4862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4922..4938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4922..4938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4935..4957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4946..4962
                     /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(4970..4976,4977..4994,4995..5000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5015..5036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5153..5170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5324..5912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5404..5423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6083..6874,6875..6943))
                     /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     6706..6725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6944..7048)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7123..7142)
                     /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 aaaaaagctg
      841 gcaccatgtc tggagaacta ccaccaaaca ttaacatcaa ggaacctcga tgggatcaaa
      901 gcactttcat tggacgagcc aatcatttct tcactgtaac tgaccccagg aacattctgt
      961 taaccaacga acaactcgag agtgcgagaa aaatagtaca tgattacagg caaggaattg
     1021 ttcctcctgg tcttacagaa aatgaattgt ggagagcaaa gtacatctat gattcagctt
     1081 ttcatcctga cactggtgag aagatgattt tgataggaag aatgtcagcc caggttccca
     1141 tgaacatgac catcacaggt tgtatgatga cgttttacag gactacgccg gctgtgctgt
     1201 tctggcagtg gattaaccag tccttcaatg ccgtcgtcaa ttacaccaac agaagtggag
     1261 acgcacccct cactgtcaat gagttgggaa cagcttacgt ttctgcaaca actggtgccg
     1321 tagcaacagc tctaggactc aatgcattga ccaagcatgt ctcaccactg ataggacgtt
     1381 ttgttccctt tgctgccgta gctgctgcta attgcattaa tattccatta atgaggcaaa
     1441 gggaactcaa agttggcatt cccgtcacgg atgagaatgg gaaccgcttg ggggagtcgg
     1501 cgaacgctgc gaaacaagcc atcacgcaag ttgtcgtgtc caggattctc atggcagccc
     1561 ctggcatggc catccctcca ttcattatga acactttgga aaagaaagcc tttttgaaga
     1621 ggttcccatg gatgagtgca cccattcaag ttgggttagt tggcttctgt ttggtgtttg
     1681 ctacacccct gtgttgtgcc ctgtttcctc agaaaagttc catgtctgtg acaagcttgg
     1741 aggccgagtt gcaagctaag atccaagaga gccatcctga attgcgacgc gtgtacttca
     1801 ataagggatt gggcgaccca gctttcttgt acaaagtggt tgatatcaag gccgccgaca
     1861 tcaccagcct gtacaagaaa gtggggtgga gccatcccca gttcgagaag ggcggaggat
     1921 ctggcggcgg aagcggcgga ggaagctggt cccaccctca gtttgagaaa ggcaccgagc
     1981 tgggcagcac catggccagc tacccctacg acgtgcccga ctacgccaag gacaacaccg
     2041 tgcccctgaa gctgatcgcc ctgctggcca acggcgagtt ccactctggc gagcagctgg
     2101 gcgagacact gggcatgagc agagccgcca tcaacaagca catccagacc ctgcgggact
     2161 ggggcgtgga cgtgttcaca gtgcccggca agggctacag cctgcccgag cccatccagc
     2221 tgctgaacgc caagcagatc ctgggccagc tggatggcgg atctgtggcc gtgctgcccg
     2281 tgatcgacag caccaaccag tatctgctcg accggatcgg cgagctgaag tccggcgacg
     2341 cctgtatcgc cgagtaccag caggccggca gaggcggcag gggcagaaag tggttcagcc
     2401 ccttcggcgc caacctgtac ctgagcatgt tctggagact ggaacagggc ccagccgccg
     2461 ctattggcct gagcctggtc atcggcatcg tgatggccga ggtgctgcgg aagctgggcg
     2521 ccgacaaagt gcgcgtgaag tggcccaacg acctgtatct gcaggaccgg aagctggccg
     2581 gcatcctggt ggaactgacc ggcaaaacag gcgacgccgc ccagatcgtg atcggcgctg
     2641 gcatcaacat ggccatgcgg cgggtggaag agagcgtggt caaccagggc tggatcaccc
     2701 tgcaggaagc cggcatcaac ctggaccgga acaccctggc cgccatgctg atcagagagc
     2761 tgagagccgc cctggaactg ttcgagcagg aaggcctggc cccctacctg agcagatggg
     2821 agaagctgga caacttcatc aaccggcccg tgaaactgat catcggcgac aaagagatct
     2881 tcggcatcag ccggggcatc gacaagcagg gggccctgct cctggaacag gacggcatca
     2941 tcaagccctg gatgggcggc gagatcagcc tgcggagcgc cgagaagtga tgactcgagt
     3001 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3061 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3121 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3181 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3241 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3301 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3361 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3421 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3481 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3541 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
     3601 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     3661 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     3721 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     3781 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     3841 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     3901 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     3961 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     4021 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     4081 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     4141 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     4201 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     4261 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     4321 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     4381 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     4441 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     4501 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     4561 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     4621 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4681 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4741 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4801 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4861 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4921 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4981 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5041 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5101 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5161 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5221 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5281 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5341 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5401 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5461 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5521 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5581 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5641 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5701 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5761 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5821 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5881 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5941 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6001 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6061 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6181 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7081 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     7141 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7201 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     7261 tagggttagg cgttttgcgc tgcttcgcga tgtacg
//
