LOCUS       sequence-212874-        6969 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 6969)
  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 6969)
  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_RAB11A
FEATURES             Location/Qualifiers
     source          1..6969
                     /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"
     protein_bind    complement(2639..2663)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2701..2718)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2707..2931
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3064..3083)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3215..3262
                     /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             3270..4292
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3270..4292
                     /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    4422..4543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4459..4478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4513..4532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4592..4608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4592..4608)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4605..4627)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4616..4632
                     /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(4640..4646,4647..4664,4665..4670))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4685..4706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4823..4840)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4994..5582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5074..5093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5753..6544,6545..6613))
                     /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     6376..6395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6614..6718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6793..6812)
                     /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 aaaagcaggc
     1981 tccaccatgg gcacccgcga cgacgagtac gactacctct ttaaagttgt ccttattgga
     2041 gattctggtg ttggaaagag taatctcctg tctcgattta ctcgaaatga gtttaatctg
     2101 gaaagcaaga gcaccattgg agtagagttt gcaacaagaa gcatccaggt tgatggaaaa
     2161 acaataaagg cacagatatg ggacacagca gggcaagagc gatatcgagc tataacatca
     2221 gcatattatc gtggagctgt aggtgcctta ttggtttatg acattgctaa acatctcaca
     2281 tatgaaaatg tagagcgatg gctgaaagaa ctgagagatc atgctgatag taacattgtt
     2341 atcatgcttg tgggcaataa gagtgatcta cgtcatctca gggcagttcc tacagatgaa
     2401 gcaagagctt ttgcagaaaa gaatggtttg tcattcattg aaacttcggc cctagactct
     2461 acaaatgtag aagctgcttt tcagacaatt ttaacagaga tttaccgcat tgtttctcag
     2521 aagcaaatgt cagacagacg cgaaaatgac atgtctccaa gcaacaatgt ggttcctatt
     2581 catgttccac caaccactga aaacaagcca aaggtgcagt gctgtcagaa catcttggac
     2641 ccagctttct tgtacaaagt ggtgacgtaa gctaggggcc cgtttaaacc cgctgatcag
     2701 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2761 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2821 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2881 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2941 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3001 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3061 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3121 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3181 aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt
     3241 ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg
     3301 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     3361 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     3421 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     3481 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     3541 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     3601 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     3661 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     3721 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     3781 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     3841 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     3901 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     3961 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     4021 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     4081 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     4141 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     4201 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     4261 gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg
     4321 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4381 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4441 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4501 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4561 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4621 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4681 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4741 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4801 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4861 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4921 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5161 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5401 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5521 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5581 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5641 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5701 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5761 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5821 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5881 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5941 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6001 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6061 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6121 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6181 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6241 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6301 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6361 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6421 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6481 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6541 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6601 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6721 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6781 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6841 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6901 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     6961 tgtacgggc
//
