LOCUS       Exported                8166 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    MAC_C_AIFM1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8166)
  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 8166)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8166
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    599..617
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    620..638
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     648..672
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    815..839
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2682..2706)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2753..2776
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2813..2836
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2867..2893
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2894..3853
                     /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(3897..3914)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3903..4127
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4260..4279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4411..4458
                     /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             4466..5488
                     /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    5618..5739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5655..5674)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5709..5728
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5788..5804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5788..5804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5801..5823)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5812..5828
                     /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(5836..5866)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5881..5902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6019..6036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6190..6778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6270..6289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6949..7809)
                     /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     7572..7591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7810..7914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7989..8008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      601 cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg
      661 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      721 gaccgatcca gcctccggac tctagcgttt aaacttaagc ttggtaccga gctcggatcc
      781 actagtccag tgtggtggaa ttctgcagat atcaacaagt ttgtacaaaa aagttggcac
      841 catgttccgg tgtggaggcc tggcggcggg tgctttgaag cagaagctgg tgcccttggt
      901 gcggaccgtg tgcgtccgaa gcccgaggca gaggaaccgg ctcccaggca acttgttcca
      961 gcgatggcat gttcctctag aactccagat gacaagacaa atggctagct ctggtgcatc
     1021 agggggcaaa atcgataatt ctgtgttagt ccttattgtg ggcttatcaa cagtaggagc
     1081 tggtgcctat gcctacaaga ctatgaaaga ggatgaaaaa agatacaatg aaagaatttc
     1141 agggttaggg ctgacaccag aacagaaaca gaaaaaggcc gcgttatctg cttcagaagg
     1201 agaggaagtt cctcaagaca aggcgccaag tcatgttcct ttcctgctaa ttggtggagg
     1261 cacagctgct tttgctgcag ccagatccat ccgggctcgg gatcctgggg ccagggtact
     1321 gattgtatct gaagatcctg agctgccgta catgcgacct cctctttcaa aagaactgtg
     1381 gttttcagat gacccaaatg tcacaaagac actgcgattc aaacagtgga atggaaaaga
     1441 gagaagcata tatttccagc caccttcttt ctatgtctct gctcaggacc tgcctcatat
     1501 tgagaatggt ggtgtggctg tcctcactgg gaagaaggta gtacagctgg atgtgagaga
     1561 caacatggtg aaacttaatg atggctctca aataacctat gaaaagtgct tgattgcaac
     1621 aggaggtact ccaagaagtc tgtctgccat tgatagggct ggagcagagg tgaagagtag
     1681 aacaacgctt ttcagaaaga ttggagactt tagaagcttg gagaagattt cacgggaagt
     1741 caaatcaatt acgattatcg gtgggggctt ccttggtagc gaactggcct gtgctcttgg
     1801 cagaaaggct cgagccttgg gcacagaagt gattcaactc ttccccgaga aaggaaatat
     1861 gggaaagatc ctccccgaat acctcagcaa ctggaccatg gaaaaagtca gacgagaggg
     1921 ggttaaggtg atgcccaatg ctattgtgca atccgttgga gtcagcagtg gcaagttact
     1981 tatcaagctg aaagacggca ggaaggtaga aactgaccac atagtggcag ctgtgggcct
     2041 ggagcccaat gttgagttgg ccaagactgg tggcctggaa atagactcag attttggtgg
     2101 cttccgggta aatgcagagc tacaagcacg ctctaacatc tgggtggcag gagatgctgc
     2161 atgcttctac gatataaagt tgggaaggag gcgggtagag caccatgatc acgctgttgt
     2221 gagtggaaga ttggctggag aaaatatgac tggagctgct aagccgtact ggcatcagtc
     2281 aatgttctgg agtgatttgg gccccgatgt tggctatgaa gctattggtc ttgtggacag
     2341 tagtttgccc acagttggtg tttttgcaaa agcaactgca caagacaacc ccaaatctgc
     2401 cacagagcag tcaggaactg gtatccgatc agagagtgag acagagtccg aggcctcaga
     2461 aattactatt cctcccagca ccccggcagt tccacaggct cccgtccagg gggaggacta
     2521 cggcaaaggt gtcatcttct acctcaggga caaagtggtc gtggggattg tgctatggaa
     2581 catctttaac cgaatgccaa tagcaaggaa gatcattaag gacggtgagc agcatgaaga
     2641 tctcaatgaa gtagccaaac tattcaacat tcatgaagac ttgccaactt tcttgtacaa
     2701 agtggttgat atcaaggccg ccgacatcac cagcctgtac aagaaagtgg ggtggagcca
     2761 tccccagttc gagaagggcg gaggatctgg cggcggaagc ggcggaggaa gctggtccca
     2821 ccctcagttt gagaaaggca ccgagctggg cagcaccatg gccagctacc cctacgacgt
     2881 gcccgactac gccaaggaca acaccgtgcc cctgaagctg atcgccctgc tggccaacgg
     2941 cgagttccac tctggcgagc agctgggcga gacactgggc atgagcagag ccgccatcaa
     3001 caagcacatc cagaccctgc gggactgggg cgtggacgtg ttcacagtgc ccggcaaggg
     3061 ctacagcctg cccgagccca tccagctgct gaacgccaag cagatcctgg gccagctgga
     3121 tggcggatct gtggccgtgc tgcccgtgat cgacagcacc aaccagtatc tgctcgaccg
     3181 gatcggcgag ctgaagtccg gcgacgcctg tatcgccgag taccagcagg ccggcagagg
     3241 cggcaggggc agaaagtggt tcagcccctt cggcgccaac ctgtacctga gcatgttctg
     3301 gagactggaa cagggcccag ccgccgctat tggcctgagc ctggtcatcg gcatcgtgat
     3361 ggccgaggtg ctgcggaagc tgggcgccga caaagtgcgc gtgaagtggc ccaacgacct
     3421 gtatctgcag gaccggaagc tggccggcat cctggtggaa ctgaccggca aaacaggcga
     3481 cgccgcccag atcgtgatcg gcgctggcat caacatggcc atgcggcggg tggaagagag
     3541 cgtggtcaac cagggctgga tcaccctgca ggaagccggc atcaacctgg accggaacac
     3601 cctggccgcc atgctgatca gagagctgag agccgccctg gaactgttcg agcaggaagg
     3661 cctggccccc tacctgagca gatgggagaa gctggacaac ttcatcaacc ggcccgtgaa
     3721 actgatcatc ggcgacaaag agatcttcgg catcagccgg ggcatcgaca agcagggggc
     3781 cctgctcctg gaacaggacg gcatcatcaa gccctggatg ggcggcgaga tcagcctgcg
     3841 gagcgccgag aagtgatgac tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3901 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3961 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4021 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4081 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     4141 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4201 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4261 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4321 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4381 acttgattag ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct
     4441 agaaagtata ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     4501 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     4561 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     4621 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     4681 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     4741 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4801 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     4861 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4921 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     4981 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     5041 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     5101 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     5161 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     5221 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     5281 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     5341 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     5401 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     5461 cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc
     5521 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5581 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5641 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5701 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5761 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5821 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5881 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5941 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7921 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     7981 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     8041 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     8101 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     8161 cgggcc
//
