LOCUS       sequence-451888-        7292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to transiently express a TurboID fusion of the protein kinase 
            MEK1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7292)
  AUTHORS   Simon JJ, Fowler DM, Maly DJ
  TITLE     Multiplexed profiling of intracellular protein abundance, activity, 
            interactions and druggability with LABEL-seq.
  JOURNAL   Nat Methods. 2024 Nov;21(11):2094-2106. doi: 
            10.1038/s41592-024-02456-7. Epub 2024 Oct 21.
  PUBMED    39433876
REFERENCE   2  (bases 1 to 7292)
  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"; doi: 
            "10.1038/s41592-024-02456-7"; journalName: "Nat Methods"; date: 
            "2024-11"; volume: "21"; issue: "11"; pages: "2094-2106"
COMMENT     Sequence Label: TurboID-MEK1
FEATURES             Location/Qualifiers
     source          1..7292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(404..1195,1196..1264))
                     /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"
     promoter        complement(1265..1369)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1444..1463)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1635..2014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2015..2218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2169..2189
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2220..2238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2241..2259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2269..2293
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2387..2396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2396..2425
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2432..3388
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     primer_bind     complement(4644..4661)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4650..4874
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5007..5026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5158..5205
                     /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             5213..6235
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5213..6235
                     /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    6365..6486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6402..6421)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6456..6475
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6535..6551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6535..6551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6548..6570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6559..6575
                     /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(6583..6589,6590..6607,6608..6613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6628..6649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6766..6783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6937..7292,1..233))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7017..7036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
       61 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
      961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1381 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     1441 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     1501 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     1561 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     1621 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     1681 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1741 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1801 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1861 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1921 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1981 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     2041 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2101 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     2161 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     2221 ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt
     2281 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
     2341 ggaccgatcc agcctccgga ctctagcgtt taaacttaag cttgccgcca ccatggaaca
     2401 aaaactcatc tcagaagagg atctcggtgg aaaagacaat actgtgcctc tgaagctgat
     2461 cgctctcctg gctaatggcg agttccatag tggcgaacag ctgggagaaa ccctgggcat
     2521 gtccagggcc gctatcaaca agcacattca gactctgcgc gactggggcg tggacgtgtt
     2581 caccgtgccc ggaaagggct actctctgcc cgagcctatc ccgctgctga acgctaaaca
     2641 gattctggga cagctggacg gcgggagcgt ggcagtcctg cctgtggtcg actccaccaa
     2701 tcagtacctg ctggatcgaa tcggcgagct gaagagtggg gatgcttgca ttgcagaata
     2761 tcagcaggca gggagaggaa gcagagggag gaaatggttc tctccttttg gagctaacct
     2821 gtacctgagt atgttttggc gcctgaagcg gggaccagca gcaatcggcc tgggcccggt
     2881 catcggaatt gtcatggcag aagcgctgcg aaagctggga gcagacaagg tgcgagtcaa
     2941 atggcccaat gacctgtatc tgcaggatag aaagctggca ggcatcctgg tggagctggc
     3001 cggaataaca ggcgatgctg cacagatcgt cattggcgcc gggattaacg tggctatgag
     3061 gcgcgtggag gaaagcgtgg tcaatcaggg ctggatcaca ctgcaggaag cagggattaa
     3121 cctggacagg aatactctgg ccgctacgct gatccgagag ctgcgggcag ccctggaact
     3181 gttcgagcag gaaggcctgg ctccatatct gccacggtgg gagaagctgg ataacttcat
     3241 caatagaccc gtgaagctga tcattgggga caaagagatt ttcgggatta gccgggggat
     3301 tgataaacag ggagccctgc tgctggaaca ggacggagtt atcaaaccct ggatgggcgg
     3361 agaaatcagt ctgcggtctg ccgaaaagct gcagggtggc agcggtagtg gatctggctc
     3421 aggaatgccc aagaagaagc cgacgcccat ccagctgaac ccggcccccg acggctctgc
     3481 agttaacggg accagctctg cggagaccaa cttggaggcc ttgcagaaga agctggagga
     3541 gctagagctt gatgagcagc agcgaaagcg ccttgaggcc tttcttaccc agaagcagaa
     3601 ggtgggagaa ctgaaggatg acgactttga gaagatcagt gagctggggg ctggcaatgg
     3661 cggtgtggtg ttcaaggtct cccacaagcc ttctggcctg gtcatggcca gaaagctaat
     3721 tcatctggag atcaaacccg caatccggaa ccagatcata agggagctgc aggttctgca
     3781 tgagtgcaac tctccgtaca tcgtgggctt ctatggtgcg ttctacagcg atggcgagat
     3841 cagtatctgc atggagcaca tggatggagg ttctctggat caagtcctga agaaagctgg
     3901 aagaattcct gaacaaattt taggaaaagt tagcattgct gtaataaaag gcctgacata
     3961 tctgagggag aagcacaaga tcatgcacag agatgtcaag ccctccaaca tcctagtcaa
     4021 ctcccgtggg gagatcaagc tctgtgactt tggggtcagc gggcagctca tcgactccat
     4081 ggccaactcc ttcgtgggca caaggtccta catgtcgcca gaaagactcc aggggactca
     4141 ttactctgtg cagtcagaca tctggagcat gggactgtct ctggtagaga tggcggttgg
     4201 gaggtatccc atccctcctc cagatgccaa ggagctggag ctgatgtttg ggtgccaggt
     4261 ggaaggagat gcggctgaga ccccacccag gccaaggacc cccgggaggc cccttagctc
     4321 atacggaatg gacagccgac ctcccatggc aatttttgag ttgttggatt acatagtcaa
     4381 cgagcctcct ccaaaactgc ccagtggagt gttcagtctg gaatttcaag attttgtgaa
     4441 taaatgctta ataaaaaacc ccgcagagag agcagatttg aagcaactca tggttcatgc
     4501 ttttatcaag agatctgatg ctgaggaagt ggattttgca ggttggctct gctccaccat
     4561 cggccttaac cagcccagca caccaaccca tgctgctggc gtctagctcg agtctagagg
     4621 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4681 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4741 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4801 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4861 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4981 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5041 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5101 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa
     5161 gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc
     5221 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     5281 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     5341 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     5401 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     5461 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     5521 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     5581 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     5641 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     5701 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     5761 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     5821 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     5881 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     5941 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     6001 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     6061 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     6121 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     6181 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     6241 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6301 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6361 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6421 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6481 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6541 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6601 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6661 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6721 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7261 aagacacgac ttatcgccac tggcagcagc ca
//
