LOCUS       sequence-212883-        6747 bp    DNA     circular SYN 14-APR-2025
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6747)
  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 6747)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_TOM22
FEATURES             Location/Qualifiers
     source          1..6747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        24..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     558..578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    609..627
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    630..648
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     658..682
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(850..1200,1201..1203,1204..1812)
                     /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             1813..1839
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1870..1893
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1930..1953
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1966..1990
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2426..2450)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2488..2505)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2494..2718
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2851..2870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3002..3049
                     /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             3057..4079
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3057..4079
                     /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    4209..4330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4246..4265)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4300..4319
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4379..4395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4379..4395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4392..4414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4403..4419
                     /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(4427..4433,4434..4451,4452..4457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4472..4493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4610..4627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4781..5369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4861..4880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5540..6331,6332..6400))
                     /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     6163..6182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6401..6505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6580..6599)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
       61 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      361 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      601 cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc
      661 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      721 aagacaccgg gaccgatcca gcctccggac tctagcgttt aaacttaagc ttggtaccga
      781 gctcggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg
      841 ctcgagacca tgaaggacaa caccgtgccc ctgaagctga tcgccctgct ggccaacggc
      901 gagttccact ctggcgagca gctgggcgag acactgggca tgagcagagc cgccatcaac
      961 aagcacatcc agaccctgcg ggactggggc gtggacgtgt tcacagtgcc cggcaagggc
     1021 tacagcctgc ccgagcccat ccagctgctg aacgccaagc agatcctggg ccagctggat
     1081 ggcggatctg tggccgtgct gcccgtgatc gacagcacca accagtatct gctcgaccgg
     1141 atcggcgagc tgaagtccgg cgacgcctgt atcgccgagt accagcaggc cggcagaggc
     1201 ggcaggggca gaaagtggtt cagccccttc ggcgccaacc tgtacctgag catgttctgg
     1261 agactggaac agggcccagc cgccgctatt ggcctgagcc tggtcatcgg catcgtgatg
     1321 gccgaggtgc tgcggaagct gggcgccgac aaagtgcgcg tgaagtggcc caacgacctg
     1381 tatctgcagg accggaagct ggccggcatc ctggtggaac tgaccggcaa aacaggcgac
     1441 gccgcccaga tcgtgatcgg cgctggcatc aacatggcca tgcggcgggt ggaagagagc
     1501 gtggtcaacc agggctggat caccctgcag gaagccggca tcaacctgga ccggaacacc
     1561 ctggccgcca tgctgatcag agagctgaga gccgccctgg aactgttcga gcaggaaggc
     1621 ctggccccct acctgagcag atgggagaag ctggacaact tcatcaaccg gcccgtgaaa
     1681 ctgatcatcg gcgacaaaga gatcttcggc atcagccggg gcatcgacaa gcagggggcc
     1741 ctgctcctgg aacaggacgg catcatcaag ccctggatgg gcggcgagat cagcctgcgg
     1801 agcgccgaga agtacccata cgatgttcct gactatgccg gtaccgagct cggatccacc
     1861 atggctagct ggagccaccc gcagttcgag aaaggtggag gttccggagg tggatcggga
     1921 ggtggatcgt ggagccaccc gcagttcgaa aaagcggccg atatcacaag tttgtacaaa
     1981 aaagcaggct ccaccatggc tgccgccgtc gctgctgccg gtgcagggga accccagtcc
     2041 ccggacgaat tgctcccgaa aggcgacgcg gagaagcctg aggaggagct ggaggaggac
     2101 gacgatgagg agctagatga gaccctgtcg gagagactat ggggcctgac ggagatgttt
     2161 ccggagaggg tccggtccgc ggccggagcc acttttgatc tttccctctt tgtggctcag
     2221 aaaatgtaca ggttttccag ggcagccttg tggattggga ccacttcctt tatgatcctg
     2281 gttcttcccg ttgtctttga gacggagaag ttgcaaatgg agcaacagca gcaactgcag
     2341 cagcggcaga tacttctagg acctaacaca gggctctcag gaggaatgcc aggggctcta
     2401 ccctcacttc ctggaaagat cttggaccca gctttcttgt acaaagtggt gacgtaagct
     2461 aggggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2521 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2581 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2641 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2701 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2761 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2821 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2881 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2941 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct
     3001 agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa
     3061 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     3121 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     3181 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     3241 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
     3301 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     3361 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     3421 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     3481 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     3541 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     3601 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     3661 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     3721 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     3781 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     3841 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     3901 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     3961 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     4021 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc
     4081 acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4141 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4201 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4261 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4321 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4381 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4441 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4501 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4561 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4621 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4681 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6481 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6541 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     6601 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     6661 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     6721 ggttaggcgt tttgcgctgc ttcgcga
//
