LOCUS       sequence-212859-        7275 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 7275)
  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 7275)
  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_FBRL
FEATURES             Location/Qualifiers
     source          1..7275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        395..598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     549..569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    600..618
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    621..639
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     649..673
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(841..1191,1192..1194,1195..1803)
                     /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             1804..1830
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1861..1884
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1921..1944
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1957..1981
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2755..2766)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     protein_bind    complement(2945..2969)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3007..3024)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3013..3237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3370..3389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3521..3568
                     /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             3576..4598
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3576..4598
                     /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    4728..4849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4765..4784)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4819..4838
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4898..4914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4898..4914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4911..4933)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4922..4938
                     /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(4946..4952,4953..4970,4971..4976))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4991..5012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5129..5146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5300..5888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5380..5399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6059..6850,6851..6919))
                     /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     6682..6701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6920..7024)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7099..7118)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      421 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      481 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      541 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      601 ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt
      661 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
      721 ggaccgatcc agcctccgga ctctagcgtt taaacttaag cttggtaccg agctcggatc
      781 cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagacc
      841 atgaaggaca acaccgtgcc cctgaagctg atcgccctgc tggccaacgg cgagttccac
      901 tctggcgagc agctgggcga gacactgggc atgagcagag ccgccatcaa caagcacatc
      961 cagaccctgc gggactgggg cgtggacgtg ttcacagtgc ccggcaaggg ctacagcctg
     1021 cccgagccca tccagctgct gaacgccaag cagatcctgg gccagctgga tggcggatct
     1081 gtggccgtgc tgcccgtgat cgacagcacc aaccagtatc tgctcgaccg gatcggcgag
     1141 ctgaagtccg gcgacgcctg tatcgccgag taccagcagg ccggcagagg cggcaggggc
     1201 agaaagtggt tcagcccctt cggcgccaac ctgtacctga gcatgttctg gagactggaa
     1261 cagggcccag ccgccgctat tggcctgagc ctggtcatcg gcatcgtgat ggccgaggtg
     1321 ctgcggaagc tgggcgccga caaagtgcgc gtgaagtggc ccaacgacct gtatctgcag
     1381 gaccggaagc tggccggcat cctggtggaa ctgaccggca aaacaggcga cgccgcccag
     1441 atcgtgatcg gcgctggcat caacatggcc atgcggcggg tggaagagag cgtggtcaac
     1501 cagggctgga tcaccctgca ggaagccggc atcaacctgg accggaacac cctggccgcc
     1561 atgctgatca gagagctgag agccgccctg gaactgttcg agcaggaagg cctggccccc
     1621 tacctgagca gatgggagaa gctggacaac ttcatcaacc ggcccgtgaa actgatcatc
     1681 ggcgacaaag agatcttcgg catcagccgg ggcatcgaca agcagggggc cctgctcctg
     1741 gaacaggacg gcatcatcaa gccctggatg ggcggcgaga tcagcctgcg gagcgccgag
     1801 aagtacccat acgatgttcc tgactatgcc ggtaccgagc tcggatccac catggctagc
     1861 tggagccacc cgcagttcga gaaaggtgga ggttccggag gtggatcggg aggtggatcg
     1921 tggagccacc cgcagttcga aaaagcggcc gatatcacaa gtttgtacaa aaaagttggc
     1981 atgaagccag gattcagtcc ccgtgggggt ggctttggcg gccgaggggg ctttggtgac
     2041 cgtggtggtc gtggaggccg agggggcttt ggcgggggcc gaggtcgagg cggaggcttt
     2101 agaggtcgtg gacgaggagg aggtggaggc ggcggcggcg gtggaggagg aggaagaggt
     2161 ggtggaggct tccattctgg tggcaaccgg ggtcgtggtc ggggaggaaa aagaggaaac
     2221 cagtcgggga agaatgtgat ggtggagccg catcggcatg agggtgtctt catttgtcga
     2281 ggaaaggaag atgcactggt caccaagaac ctggtccctg gggaatcagt ttatggagag
     2341 aagagagtct cgatttcgga aggagatgac aaaattgagt accgagcctg gaaccccttc
     2401 cgctccaagc tagcagcagc aatcctgggt ggtgtggacc agatccacat caaaccgggg
     2461 gctaaggttc tctacctcgg ggctgcctcg ggcaccacgg tctcccatgt ctctgacatc
     2521 gttggtccgg atggtctagt ctatgcagtc gagttctccc accgctctgg ccgtgacctc
     2581 attaacttgg ccaagaagag gaccaacatc attcctgtga tcgaggatgc tcgacaccca
     2641 cacaaatacc gcatgctcat cgcaatggtg gatgtgatct ttgctgatgt ggcccagcca
     2701 gaccagaccc ggattgtggc cctgaatgcc cacaccttcc tgcgtaatgg aggacacttt
     2761 gtgatttcca ttaaggccaa ctgcattgac tccacagcct cagccgaggc cgtgtttgcc
     2821 tccgaagtga aaaagatgca acaggagaac atgaagccgc aggagcagtt gacccttgag
     2881 ccatatgaaa gagaccatgc cgtggtcgtg ggagtgtaca ggccaccccc caaggtgaag
     2941 aactgcccaa ctttcttgta caaagtggtg acgtaagcta ggggcccgtt taaacccgct
     3001 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3061 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3121 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3181 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3241 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3481 accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta ttccgaagtt
     3541 cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc accgcgacgt
     3601 ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg
     3661 agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg
     3721 taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg
     3781 ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt
     3841 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
     3901 ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga
     3961 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
     4021 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
     4081 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
     4141 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
     4201 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
     4261 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
     4321 agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg
     4381 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
     4441 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
     4501 cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
     4561 accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt
     4621 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4681 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4741 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4801 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4861 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4921 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4981 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5041 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5101 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5161 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5221 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5281 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5341 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5401 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5461 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5521 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5581 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5641 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5701 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5761 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5821 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5881 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5941 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6001 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6061 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6121 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6181 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6241 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6301 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6361 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6421 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6481 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6541 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6601 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6661 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6721 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6781 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6841 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6901 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6961 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7021 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7081 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     7141 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     7201 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     7261 tcgcgatgta cgggc
//
