LOCUS       sequence-212735-        7518 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 7518)
  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 7518)
  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_ARP2
FEATURES             Location/Qualifiers
     source          1..7518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        22..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    628..646
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     656..680
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    823..847
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2042..2066)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2113..2136
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2173..2196
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2227..2253
                     /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             2227..2253
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2254..2601,2602..2604,2605..3213)
                     /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(3257..3274)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3263..3487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3620..3639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3771..3818
                     /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             3826..4848
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3826..4848
                     /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    4978..5099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5015..5034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5069..5088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5148..5164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5148..5164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5161..5183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5172..5188
                     /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(5196..5202,5203..5220,5221..5226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5241..5262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5379..5396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5550..6138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5630..5649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6309..7100,7101..7169))
                     /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     6932..6951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7170..7274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7349..7368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctctccc tatcagtgat agagatctcc ctatcagtga tagagatcgt cgacgagctc
      661 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
      721 gacaccggga ccgatccagc ctccggactc tagcgtttaa acttaagctt ggtaccgagc
      781 tcggatccac tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa
      841 gcaggctcca ccatggacag ccagggcagg aaggtggtgg tgtgcgacaa cggcaccggg
      901 tttgtgaagt gtggatatgc aggctctaac tttccagaac acatcttccc agctttggtt
      961 ggaagaccta ttatcagatc aaccaccaaa gtgggaaaca ttgaaatcaa ggatcttatg
     1021 gttggtgatg aggcaagtga attacgatca acgttagaag ttaactaccc tatggaaaat
     1081 ggcatagtac gaaattggga tgacatgaaa cacctgtggg actacacatt tggaccagag
     1141 aaacttaata tagataccag aaattgtaaa atcttactca cagaacctcc tatgaaccca
     1201 accaaaaaca gagagaagat tgtagaggta atgtttgaaa cttaccagtt ttccggtgta
     1261 tatgtagcca tccaggcagt tctgactttg tacgctcaag gtttattgac tggtgtagtg
     1321 gtagactctg gagatggtgt gactcacatt tgcccagtat atgaaagctt ttctctccct
     1381 catcttacca ggagactgga tattgctggg agggatataa ctagatatct tatcaagcta
     1441 cttctgttgc gaggatacgc cttcaaccac tctgctgatt ttgaaacggt tcgcatgatt
     1501 aaagaaaaac tgtgttacgt gggatataat attgagcaag agcagaaact ggccttagaa
     1561 accacagtat tagttgaatc ttatacactc ccagatggac gtatcatcaa agttggggga
     1621 gagagatttg aagcaccaga agctttattt cagcctcact tgatcaatgt tgaaggagtt
     1681 ggtgttgctg aattgctttt taacacaatt caggcagctg acattgatac cagatctgaa
     1741 ttctacaaac acattgtgct ttctggaggg tctactatgt atcctggcct gccatcacgg
     1801 ttggaacgag aacttaaaca gctttactta gaacgagttt tgaagggtga tgtggaaaaa
     1861 ctttctaaat ttaagatccg cattgaagac ccaccccgca gaaagcacat ggtattcctg
     1921 ggtggtgcag ttctagcgga tatcatgaaa gacaaagaca acttttggat gacccgacaa
     1981 gagtaccaag aaaaaggtgt ccgtgtgcta gagaaacttg gtgtgactgt tcgatggatc
     2041 cacccagctt tcttgtacaa agtggttgat atcaaggccg ccgacatcac cagcctgtac
     2101 aagaaagtgg ggtggagcca tccccagttc gagaagggcg gaggatctgg cggcggaagc
     2161 ggcggaggaa gctggtccca ccctcagttt gagaaaggca ccgagctggg cagcaccatg
     2221 gccagctacc cctacgacgt gcccgactac gccaaggaca acaccgtgcc cctgaagctg
     2281 atcgccctgc tggccaacgg cgagttccac tctggcgagc agctgggcga gacactgggc
     2341 atgagcagag ccgccatcaa caagcacatc cagaccctgc gggactgggg cgtggacgtg
     2401 ttcacagtgc ccggcaaggg ctacagcctg cccgagccca tccagctgct gaacgccaag
     2461 cagatcctgg gccagctgga tggcggatct gtggccgtgc tgcccgtgat cgacagcacc
     2521 aaccagtatc tgctcgaccg gatcggcgag ctgaagtccg gcgacgcctg tatcgccgag
     2581 taccagcagg ccggcagagg cggcaggggc agaaagtggt tcagcccctt cggcgccaac
     2641 ctgtacctga gcatgttctg gagactggaa cagggcccag ccgccgctat tggcctgagc
     2701 ctggtcatcg gcatcgtgat ggccgaggtg ctgcggaagc tgggcgccga caaagtgcgc
     2761 gtgaagtggc ccaacgacct gtatctgcag gaccggaagc tggccggcat cctggtggaa
     2821 ctgaccggca aaacaggcga cgccgcccag atcgtgatcg gcgctggcat caacatggcc
     2881 atgcggcggg tggaagagag cgtggtcaac cagggctgga tcaccctgca ggaagccggc
     2941 atcaacctgg accggaacac cctggccgcc atgctgatca gagagctgag agccgccctg
     3001 gaactgttcg agcaggaagg cctggccccc tacctgagca gatgggagaa gctggacaac
     3061 ttcatcaacc ggcccgtgaa actgatcatc ggcgacaaag agatcttcgg catcagccgg
     3121 ggcatcgaca agcagggggc cctgctcctg gaacaggacg gcatcatcaa gccctggatg
     3181 ggcggcgaga tcagcctgcg gagcgccgag aagtgatgac tcgagtctag agggcccgtt
     3241 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3301 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3361 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3421 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3481 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3541 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3601 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3661 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3721 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta
     3781 ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc
     3841 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
     3901 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     3961 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     4021 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc
     4081 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     4141 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     4201 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     4261 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     4321 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     4381 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     4441 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     4501 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     4561 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     4621 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     4681 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     4741 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     4801 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga
     4861 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4921 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4981 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5041 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5101 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5161 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5221 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5281 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5341 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5401 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7321 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7381 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7441 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     7501 ttgcgctgct tcgcgatg
//
