LOCUS       Exported                8091 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    MAC_C_EZRI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8091)
  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 8091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8091
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(396..420)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             467..490
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             527..550
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             581..607
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             608..1567
                     /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(1611..1628)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1617..1841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1974..1993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2125..2172
                     /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             2180..3202
                     /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    3332..3453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3369..3388)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3423..3442
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3502..3518)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3502..3518)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3515..3537)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3526..3542
                     /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(3550..3580)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3595..3616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3733..3750)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3904..4492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3984..4003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4663..5523)
                     /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     5286..5305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5524..5628)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5703..5722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5894..6273
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6274..6477
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6428..6448
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6479..6497
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6500..6518
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6528..6552
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6695..6719
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ggtgaagacc aaggaggagc tgcacctggt gatgacagca cccccgcccc caccaccccc
       61 cgtgtacgag ccggtgagct accatgtcca ggagagcttg caggatgagg gcgcagagcc
      121 cacgggctac agcgcggagc tgtctagtga gggcatccgg gatgaccgca atgaggagaa
      181 gcgcatcact gaggcagaga agaacgagcg tgtgcagcgg cagctgctga cgctgagcag
      241 cgagctgtcc caggcccgag atgagaataa gaggacccac aatgacatca tccacaacga
      301 gaacatgagg caaggccggg acaagtacaa gacgctgcgg cagatccggc agggcaacac
      361 caagcagcgc atcgacgagt tcgaggccct gttggaccca gctttcttgt acaaagtggt
      421 tgatatcaag gccgccgaca tcaccagcct gtacaagaaa gtggggtgga gccatcccca
      481 gttcgagaag ggcggaggat ctggcggcgg aagcggcgga ggaagctggt cccaccctca
      541 gtttgagaaa ggcaccgagc tgggcagcac catggccagc tacccctacg acgtgcccga
      601 ctacgccaag gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca acggcgagtt
      661 ccactctggc gagcagctgg gcgagacact gggcatgagc agagccgcca tcaacaagca
      721 catccagacc ctgcgggact ggggcgtgga cgtgttcaca gtgcccggca agggctacag
      781 cctgcccgag cccatccagc tgctgaacgc caagcagatc ctgggccagc tggatggcgg
      841 atctgtggcc gtgctgcccg tgatcgacag caccaaccag tatctgctcg accggatcgg
      901 cgagctgaag tccggcgacg cctgtatcgc cgagtaccag caggccggca gaggcggcag
      961 gggcagaaag tggttcagcc ccttcggcgc caacctgtac ctgagcatgt tctggagact
     1021 ggaacagggc ccagccgccg ctattggcct gagcctggtc atcggcatcg tgatggccga
     1081 ggtgctgcgg aagctgggcg ccgacaaagt gcgcgtgaag tggcccaacg acctgtatct
     1141 gcaggaccgg aagctggccg gcatcctggt ggaactgacc ggcaaaacag gcgacgccgc
     1201 ccagatcgtg atcggcgctg gcatcaacat ggccatgcgg cgggtggaag agagcgtggt
     1261 caaccagggc tggatcaccc tgcaggaagc cggcatcaac ctggaccgga acaccctggc
     1321 cgccatgctg atcagagagc tgagagccgc cctggaactg ttcgagcagg aaggcctggc
     1381 cccctacctg agcagatggg agaagctgga caacttcatc aaccggcccg tgaaactgat
     1441 catcggcgac aaagagatct tcggcatcag ccggggcatc gacaagcagg gggccctgct
     1501 cctggaacag gacggcatca tcaagccctg gatgggcggc gagatcagcc tgcggagcgc
     1561 cgagaagtga tgactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     1621 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1681 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1741 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1801 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1861 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1921 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1981 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2041 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2101 ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag
     2161 tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     2221 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     2281 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     2341 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     2401 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
     2461 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     2521 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     2581 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     2641 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     2701 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     2761 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     2821 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     2881 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     2941 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     3001 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     3061 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     3121 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     3181 tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta
     3241 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3301 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3361 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3421 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3481 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3541 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3601 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3661 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3721 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3781 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3841 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3901 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3961 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4021 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4081 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4141 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4201 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4261 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4321 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4381 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4441 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4501 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4561 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4621 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4681 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4741 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4801 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4861 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4921 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4981 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5041 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5101 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5161 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5221 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5281 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5341 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5401 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5461 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5581 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5641 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     5701 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     5761 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     5821 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     5881 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     5941 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     6001 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     6061 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     6121 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     6181 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6241 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     6301 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     6361 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     6421 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     6481 cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg
     6541 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     6601 gaccgatcca gcctccggac tctagcgttt aaacttaagc ttggtaccga gctcggatcc
     6661 actagtccag tgtggtggaa ttctgcagat atcaacaagt ttgtacaaaa aagcaggctc
     6721 caccatgccg aaaccaatca atgtccgagt taccaccatg gatgcagagc tggagtttgc
     6781 aatccagcca aatacaactg gaaaacagct ttttgatcag gtggtaaaga ctatcggcct
     6841 ccgggaagtg tggtactttg gcctccacta tgtggataat aaaggatttc ctacctggct
     6901 gaagctggat aagaaggtgt ctgcccagga ggtcaggaag gagaatcccc tccagttcaa
     6961 gttccgggcc aagttctacc ctgaagatgt ggctgaggag ctcatccagg acatcaccca
     7021 gaaacttttc ttcctccaag tgaaggaagg aatccttagc gatgagatct actgcccccc
     7081 tgagactgcc gtgctcttgg ggtcctacgc tgtgcaggcc aagtttgggg actacaacaa
     7141 agaagtgcac aagtctgggt acctcagctc tgagcggctg atccctcaaa gagtgatgga
     7201 ccagcacaaa cttaccaggg accagtggga ggaccggatc caggtgtggc atgcggaaca
     7261 ccgtgggatg ctcaaagata atgctatgtt ggaatacctg aagattgctc aggacctgga
     7321 aatgtatgga atcaactatt tcgagataaa aaacaagaaa ggaacagacc tttggcttgg
     7381 agttgatgcc cttggactga atatttatga gaaagatgat aagttaaccc caaagattgg
     7441 ctttccttgg agtgaaatca ggaacatctc tttcaatgac aaaaagtttg tcattaaacc
     7501 catcgacaag aaggcacctg actttgtgtt ttatgcccca cgtctgagaa tcaacaagcg
     7561 gatcctgcag ctctgcatgg gcaaccatga gttgtatatg cgccgcagga agcctgacac
     7621 catcgaggtg cagcagatga aggcccaggc ccgggaggag aagcatcaga agcagctgga
     7681 gcggcaacag ctggaaacag agaagaaaag gagagaaacc gtggagagag agaaagagca
     7741 gatgatgcgc gagaaggagg agttgatgct gcggctgcag gactatgagg agaagacaaa
     7801 gaaggcagag agagagctct cggagcagat tcagagggcc ctgcagctgg aggaggagag
     7861 gaagcgggca caggaggagg ccgagcgcct agaggctgac cgtatggctg cactgcgggc
     7921 taaggaggag ctggagagac aggcggtgga tcagataaag agccaggagc agctggctgc
     7981 ggagcttgca gaatacactg ccaagattgc cctcctggaa gaggcgcgga ggcgcaagga
     8041 ggatgaagtt gaagagtggc agcacagggc caaagaagcc caggatgacc t
//
