LOCUS       sequence-212848-        7437 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 7437)
  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 7437)
  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_ARPC1B
FEATURES             Location/Qualifiers
     source          1..7437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     550..570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    601..619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    622..640
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     650..674
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(842..1192,1193..1195,1196..1804)
                     /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             1805..1831
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1862..1885
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1922..1945
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1958..1982
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3108..3132)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3170..3187)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3176..3400
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3533..3552)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3684..3731
                     /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             3739..4761
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3739..4761
                     /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    4891..5012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4928..4947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4982..5001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5061..5077)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5061..5077)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5074..5096)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5085..5101
                     /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(5109..5115,5116..5133,5134..5139))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5154..5175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5292..5309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5463..6051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5543..5562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6222..7013,7014..7082))
                     /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     6845..6864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7083..7187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7262..7281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      601 tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga gctcgtttag
      661 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      721 gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc gagctcggat
      781 ccactagtcc agtgtggtgg aattctgcag atatccagca cagtggcggc cgctcgagac
      841 catgaaggac aacaccgtgc ccctgaagct gatcgccctg ctggccaacg gcgagttcca
      901 ctctggcgag cagctgggcg agacactggg catgagcaga gccgccatca acaagcacat
      961 ccagaccctg cgggactggg gcgtggacgt gttcacagtg cccggcaagg gctacagcct
     1021 gcccgagccc atccagctgc tgaacgccaa gcagatcctg ggccagctgg atggcggatc
     1081 tgtggccgtg ctgcccgtga tcgacagcac caaccagtat ctgctcgacc ggatcggcga
     1141 gctgaagtcc ggcgacgcct gtatcgccga gtaccagcag gccggcagag gcggcagggg
     1201 cagaaagtgg ttcagcccct tcggcgccaa cctgtacctg agcatgttct ggagactgga
     1261 acagggccca gccgccgcta ttggcctgag cctggtcatc ggcatcgtga tggccgaggt
     1321 gctgcggaag ctgggcgccg acaaagtgcg cgtgaagtgg cccaacgacc tgtatctgca
     1381 ggaccggaag ctggccggca tcctggtgga actgaccggc aaaacaggcg acgccgccca
     1441 gatcgtgatc ggcgctggca tcaacatggc catgcggcgg gtggaagaga gcgtggtcaa
     1501 ccagggctgg atcaccctgc aggaagccgg catcaacctg gaccggaaca ccctggccgc
     1561 catgctgatc agagagctga gagccgccct ggaactgttc gagcaggaag gcctggcccc
     1621 ctacctgagc agatgggaga agctggacaa cttcatcaac cggcccgtga aactgatcat
     1681 cggcgacaaa gagatcttcg gcatcagccg gggcatcgac aagcaggggg ccctgctcct
     1741 ggaacaggac ggcatcatca agccctggat gggcggcgag atcagcctgc ggagcgccga
     1801 gaagtaccca tacgatgttc ctgactatgc cggtaccgag ctcggatcca ccatggctag
     1861 ctggagccac ccgcagttcg agaaaggtgg aggttccgga ggtggatcgg gaggtggatc
     1921 gtggagccac ccgcagttcg aaaaagcggc cgatatcaca agtttgtaca aaaaagcagg
     1981 ctccaccatg gcctaccaca gcttcctggt ggagcccatc agctgccacg cctggaacaa
     2041 ggaccgcacc cagattgcca tctgccccaa caaccatgag gtgcatatct atgaaaagag
     2101 cggtgccaaa tggaccaagg tgcacgagct caaggagcac aacgggcagg tgacaggcat
     2161 cgactgggcc cccgagagta accgtattgt gacctgcggc acagaccgca acgcctacgt
     2221 gtggacgctg aagggccgca catggaagcc cacgctggtc atcctgcgga tcaaccgggc
     2281 tgcccgctgc gtgcgctggg cccccaacga gaacaagttt gctgtgggca gcggctctcg
     2341 tgtgatctcc atctgttatt tcgagcagga gaatgactgg tgggtttgca agcacatcaa
     2401 gaagcccatc cgctccaccg tcctcagcct ggactggcac cccaacaatg tgctgctggc
     2461 tgccggctcc tgtgacttca agtgtcggat cttttcagcc tacatcaagg aggtggagga
     2521 acggccggca cccaccccgt ggggctccaa gatgcccttt ggggaactga tgttcgaatc
     2581 cagcagtagc tgcggctggg tacatggcgt ctgtttctca gccagcggga gccgcgtggc
     2641 ctgggtaagc cacgacagca ccgtctgcct ggctgatgcc gacaagaaga tggccgtcgc
     2701 gactctggcc tctgaaacac taccactgct ggcgctgacc ttcatcacag acaacagcct
     2761 ggtggcagcg ggccacgact gcttcccggt gctgttcacc tatgacgccg ccgcggggat
     2821 gctgagcttc ggcgggcggc tggacgttcc taagcagagc tcgcagcgtg gcttgacggc
     2881 ccgcgagcgc ttccagaacc tggacaagaa ggcgagctcc gagggtggca cggctgcggg
     2941 cgcgggccta gactcgctgc acaagaacag cgtcagccag atctcggtgc tcagcggcgg
     3001 caaggccaag tgctcgcagt tctgcaccac tggcatggat ggcggcatga gtatctggga
     3061 tgtgaagagc ttggagtcag ccttgaagga cctcaagatc aaattggacc cagctttctt
     3121 gtacaaagtg gtgacgtaag ctaggggccc gtttaaaccc gctgatcagc ctcgactgtg
     3181 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3241 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3301 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3361 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3421 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3481 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3541 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3601 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3661 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt
     3721 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     3781 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     3841 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     3901 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     3961 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     4021 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     4081 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     4141 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     4201 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     4261 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     4321 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     4381 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     4441 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     4501 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     4561 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     4621 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     4681 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     4741 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat
     4801 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4861 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4921 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4981 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5041 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5101 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5161 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5221 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5281 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5461 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5881 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6241 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6301 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6361 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6421 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6481 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6541 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6601 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6661 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6721 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6781 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6841 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6901 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6961 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7021 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7081 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7141 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7201 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     7261 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     7321 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     7381 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacggg
//
