LOCUS       sequence-392348-        8276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses LOV-Turbo in the yeast cytosol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8276)
  AUTHORS   Lee SY, Cheah JS, Zhao B, Xu C, Roh H, Kim CK, Cho KF, Udeshi ND, 
            Carr SA, Ting AY
  TITLE     Engineered allostery in light-regulated LOV-Turbo enables precise 
            spatiotemporal control of proximity labeling in living cells.
  JOURNAL   Nat Methods. 2023 May 15. doi: 10.1038/s41592-023-01880-5.
  PUBMED    37188954
REFERENCE   2  (bases 1 to 8276)
  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 
            Methods. 2023 May 15. doi: 10.1038/s41592-023-01880-5."
COMMENT     Sequence Label: LOV-Turbo-V5_pRS415
FEATURES             Location/Qualifiers
     source          1..8276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1221..1662
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1221..1338
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1633..1656
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3105..3146
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      3159..3406
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3175..3193)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3424..3443)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3425..3443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3453..3470)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3453..3469)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3462..3484)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3610..4065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3697..3716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3907..3928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4366..4773
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4774..5868
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(6362..6381)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6481..6503
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6541..6559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6601..7104
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7141..7245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7246..7314,7315..8106)
                     /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     complement(7464..7483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctccg caaagaagtt taataatcat attacatggc
     1081 attaccacca tatacatatc catatctaat cttacttata tgttgtggaa atgtaaagag
     1141 ccccattatc ttagcctaaa aaaaccttct ctttggaact ttcagtaata cgcttaactg
     1201 ctcattgcta tattgaagta cggattagaa gccgccgagc gggtgacagc cctccgaagg
     1261 aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc
     1321 tcgcgccgca ctgctccgaa caataaagat tctacaatac tagcttttat ggttatgaag
     1381 aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa tgaacgaatc aaattaacaa
     1441 ccataggatg ataatgcgat tagtttttta gccttatttc tggggtaatt aatcagcgaa
     1501 gcgatgattt ttgatctatt aacagatata taaatgcaaa aactgcataa ccactttaac
     1561 taatactttc aacattttcg gtttgtatta cttcttattc aaatgtaata aaagtatcaa
     1621 caaaaaattg ttaatatacc tctatacttt aacgtcaagg agaaaaaacc ccggatcgaa
     1681 ttctacttca tacattttca attaagatgg ctcgggatcc accggtcgcc accatgaaag
     1741 acaacacagt gcctctgaag ctgatcgccc tcctggccaa tggagagttc cacagcggcg
     1801 agcagctggg cgaaaccctg ggcatgagca gagccgctat caacaagcac atccagaccc
     1861 ttcgcgactg gggcgtcgac gtgttcagcg tgcccggcaa gggctacagc ctgcctgagc
     1921 ctatccccct gctgaacgcc aagcagattc agggccagct ggaatttaga gccaccaccc
     1981 tggaaagaat cgagaagtct tttgtgatca ccgaccccag actgcctgat aatcctatca
     2041 tcttcgtgtc tgatagcttc ctgcagctca ccgagtattc tcgggaagag atcctcggaa
     2101 gaaattgcag attcctgcag ggcccagaga cagatagagc cacagtgcgg aagatcagag
     2161 atgccatcga caaccagaca gaggtgaccg tgcagctgat caactacacc aagtccggca
     2221 aaaagttctg gaacgttttc catctccaac ctatgagaga ctacaagggc gacgtgcagt
     2281 acttcatcgg agttcagctg gacggcaccg agagactgca cggcgccgca gagcgggaag
     2341 ctgtgtgcct ggtcaagaag accgccttcc agatcgccgt ggctgtgctg cccgtggtgg
     2401 acagcacaaa ccagtacctg ctggacagaa tcggcgagct gaagtccggc gacgcttgta
     2461 tcgccgaata ccagcaggcc ggcagaggca gcagaggccg gaaatggttc agccccttcg
     2521 gcgccaacct gtacctgagc atgttttggc ggctgaaaag aggccctgcc gccacaggcc
     2581 tgggccccgt gataggaatc gtgatggccg aggctctgag gaagctgggt gtggacaaag
     2641 tgcgggtgaa gtggcctaac gacctgtacc tgcaggacag aaagctggcc ggcatcctgg
     2701 tggaactggc tggcatcacc ggcgacgccg ctcagattgt gatcggcgct ggcatcaatg
     2761 tggccatgag acgcgtggaa gagagcgtgg tcaaccaggg ctggatcaca ctgcaagagg
     2821 ccggtgtgaa cctggatagg aacaccctgg ccgccaccct gatcagagag ctgagagccg
     2881 ctctggaatt gtttgagcag gagggcctgg ccccttacct gccacggtgg gagaaactgg
     2941 ataacttcat caacaggcct gtgaagctga tcatcggcga taaggaaatc ctgggaatca
     3001 gccggggcat cgacaagcag ggagccctgc tgctggaaca agatggagtg attaagcctt
     3061 ggatgggcgg agaaatcagc ctgcggtccg ccgagaaggc tagcggcaag cccatcccca
     3121 accccctgct gggcctggac agcacctaat agctcgagtc atgtaattag ttatgtcacg
     3181 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     3241 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     3301 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     3361 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat
     3421 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     3481 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     3541 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3601 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3661 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3721 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3781 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3841 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3901 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3961 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4021 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     4081 tctccttacg catctgtgcg gtatttcaca ccgcatatcg accggtcgag gagaacttct
     4141 agtatatcta catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc
     4201 aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa
     4261 aacgttatat ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc
     4321 gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact
     4381 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     4441 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     4501 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     4561 ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa
     4621 atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt gttttttcca
     4681 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     4741 tatatatata tatttcaagg atataccatt ctaatgtctg cccctaagaa gatcgtcgtt
     4801 ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt tcttaaagct
     4861 atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat tggtggtgct
     4921 gctatcgatg ctacaggtgt tccacttcca gatgaggcgc tggaagcctc caagaaggct
     4981 gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg gtaccggtag tgttagacct
     5041 gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt aagaccatgt
     5101 aactttgcat ccgactctct tttagactta tctccaatca agccacaatt tgctaaaggt
     5161 actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa gagaaaggaa
     5221 gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga agtgcaaaga
     5281 atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc tatttggtcc
     5341 ttggataaag ctaatgtttt ggcctcttca agattatgga gaaaaactgt ggaggaaacc
     5401 atcaagaacg aattccctac attgaaggtt caacatcaat tgattgattc tgccgccatg
     5461 atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa catgtttggt
     5521 gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt gccatctgcg
     5581 tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc atgccacggt
     5641 tctgctccag atttgccaaa gaataaggtc aaccctatcg ccactatctt gtctgctgca
     5701 atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga agatgcagtt
     5761 aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa cagtaccacc
     5821 gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt
     5881 tttttatgat atttgtacat aaactttata aatgaaattc ataatagaaa cgacacgaaa
     5941 ttacaaaatg gaatatgttc atagggtaga cgaaactata tacgcaatct acatacattt
     6001 atcaagaagg agaaaaagga ggatgtaaag gaatacaggt aagcaaattg atactaatgg
     6061 ctcaacgtga taaggaaaaa gaattgcact ttaacattaa tattgacaag gaggagggca
     6121 ccacacaaaa agttaggtgt aacagaaaat catgaaacta tgattcctaa tttatatatt
     6181 ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc
     6241 atttgatgga gtttaagtca ataccttctt gaaccatttc ccataatggt gaaagttccc
     6301 tcaagaattt tactctgtca gaaacggcct taacgacgta gtcgatatgg tgcactctca
     6361 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6421 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6481 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6541 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttaggacg
     6601 gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa
     6661 tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga
     6721 aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca
     6781 aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat agatatacat
     6841 tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag
     6901 acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt
     6961 atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct
     7021 ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt
     7081 ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg
     7141 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7201 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7261 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7321 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     7381 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7441 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7501 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     7561 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7621 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7681 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7741 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7801 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7861 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7921 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7981 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8041 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8101 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8161 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8221 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
