LOCUS       sequence-212412-        6873 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 6873)
  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 6873)
  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_PTH2
FEATURES             Location/Qualifiers
     source          1..6873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        20..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     554..574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    605..623
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    626..644
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     654..678
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    821..845
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1395..1419)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1466..1489
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1526..1549
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1580..1606
                     /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             1580..1606
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1607..1954,1955..1957,1958..2566)
                     /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(2610..2627)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2616..2840
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2973..2992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3124..3171
                     /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             3179..4201
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3179..4201
                     /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    4331..4452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4368..4387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4422..4441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4501..4517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4501..4517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4514..4536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4525..4541
                     /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(4549..4555,4556..4573,4574..4579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4594..4615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4732..4749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4903..5491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4983..5002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5662..6453,6454..6522))
                     /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     6285..6304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6523..6627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6702..6721)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
       61 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      121 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      181 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      241 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      301 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      361 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      421 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      481 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      541 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      601 gctctcccta tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt
      661 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      721 caccgggacc gatccagcct ccggactcta gcgtttaaac ttaagcttgg taccgagctc
      781 ggatccacta gtccagtgtg gtggaattct gcagatatca acaagtttgt acaaaaaagc
      841 aggctccacc atgccctcca aatccttggt tatggaatat ttggctcatc ccagtacact
      901 cggcttggct gttggagttg cttgtggcat gtgcctgggc tggagccttc gagtatgctt
      961 tgggatgctc cccaaaagca agacgagcaa gacacacaca gatactgaaa gtgaagcaag
     1021 catcttggga gacagcgggg agtacaagat gattcttgtg gttcgaaatg acttaaagat
     1081 gggaaaaggg aaagtggctg cccagtgctc tcatgctgct gtttcagcct acaagcagat
     1141 tcaaagaaga aatcctgaaa tgctcaaaca atgggaatac tgtggccagc ccaaggtggt
     1201 ggtcaaagct cctgatgaag aaaccctgat tgcattattg gcccatgcaa aaatgctggg
     1261 actgactgta agtttaattc aagatgctgg acgtactcag attgcaccag gctctcaaac
     1321 tgtcctaggg attgggccag gaccagcaga cctaattgac aaagtcactg gtcacctaaa
     1381 actttactgg atccacccag ctttcttgta caaagtggtt gatatcaagg ccgccgacat
     1441 caccagcctg tacaagaaag tggggtggag ccatccccag ttcgagaagg gcggaggatc
     1501 tggcggcgga agcggcggag gaagctggtc ccaccctcag tttgagaaag gcaccgagct
     1561 gggcagcacc atggccagct acccctacga cgtgcccgac tacgccaagg acaacaccgt
     1621 gcccctgaag ctgatcgccc tgctggccaa cggcgagttc cactctggcg agcagctggg
     1681 cgagacactg ggcatgagca gagccgccat caacaagcac atccagaccc tgcgggactg
     1741 gggcgtggac gtgttcacag tgcccggcaa gggctacagc ctgcccgagc ccatccagct
     1801 gctgaacgcc aagcagatcc tgggccagct ggatggcgga tctgtggccg tgctgcccgt
     1861 gatcgacagc accaaccagt atctgctcga ccggatcggc gagctgaagt ccggcgacgc
     1921 ctgtatcgcc gagtaccagc aggccggcag aggcggcagg ggcagaaagt ggttcagccc
     1981 cttcggcgcc aacctgtacc tgagcatgtt ctggagactg gaacagggcc cagccgccgc
     2041 tattggcctg agcctggtca tcggcatcgt gatggccgag gtgctgcgga agctgggcgc
     2101 cgacaaagtg cgcgtgaagt ggcccaacga cctgtatctg caggaccgga agctggccgg
     2161 catcctggtg gaactgaccg gcaaaacagg cgacgccgcc cagatcgtga tcggcgctgg
     2221 catcaacatg gccatgcggc gggtggaaga gagcgtggtc aaccagggct ggatcaccct
     2281 gcaggaagcc ggcatcaacc tggaccggaa caccctggcc gccatgctga tcagagagct
     2341 gagagccgcc ctggaactgt tcgagcagga aggcctggcc ccctacctga gcagatggga
     2401 gaagctggac aacttcatca accggcccgt gaaactgatc atcggcgaca aagagatctt
     2461 cggcatcagc cggggcatcg acaagcaggg ggccctgctc ctggaacagg acggcatcat
     2521 caagccctgg atgggcggcg agatcagcct gcggagcgcc gagaagtgat gactcgagtc
     2581 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2641 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2701 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2761 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2821 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2881 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2941 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3001 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3061 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     3121 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     3181 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3241 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3301 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3361 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3421 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3481 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3541 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3601 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3661 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3721 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     3781 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     3841 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     3901 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     3961 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4021 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4081 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4141 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4201 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4261 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4321 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4381 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4441 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4501 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4561 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4621 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4681 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4741 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4801 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4861 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4921 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4981 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5041 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5101 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5161 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5221 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5281 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5341 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5401 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5461 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5521 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5581 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5641 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5701 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5761 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5821 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5881 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5941 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6001 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6061 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6121 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6181 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6241 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6301 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6361 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6421 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6481 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6541 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6601 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     6661 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     6721 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     6781 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     6841 agggttaggc gttttgcgct gcttcgcgat gta
//
