LOCUS       sequence-42500-e        6492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Mid2(SS/TM), containing only the signal sequence and 
            transmembrane helix, fused to GFP and LOVpep(K533∆, high-affinity 
            epitope)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6492)
  AUTHORS   Strickland D, Lin Y, Wagner E, Hope CM, Zayner J, Antoniou C, 
            Sosnick TR, Weiss EL, Glotzer M
  TITLE     TULIPs: tunable, light-controlled interacting protein tags for cell 
            biology.
  JOURNAL   Nat Methods. 2012 Mar 4. doi: 10.1038/nmeth.1904.
  PUBMED    22388287
REFERENCE   2  (bases 1 to 6492)
  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. 2012 Mar 4. doi: 10.1038/nmeth.1904."
COMMENT     Sequence Label: pDS240
FEATURES             Location/Qualifiers
     source          1..6492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..463
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    22..139
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     434..457
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             792..1505
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(822..850)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1445..1465
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1977..2224
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1993..2011)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2249..2265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2249..2265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2262..2284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2273..2289
                     /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(2297..2303,2304..2321,2322..2327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2342..2363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2480..2497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2651..3239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2731..2750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3410..4201,4202..4270))
                     /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     4033..4052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4271..4375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4443..4461
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4463..4867
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4880..5974
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(6115..6137)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6237..6256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6450..6472
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6464..6481
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6465..6481
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgtttaacgg cgcgcctagt acggattaga agccgccgag cgggtgacag ccctccgaag
       61 gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc
      121 ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa
      181 gaggaaaaat tggcagtaac ctggccccac aaaccttcaa atgaacgaat caaattaaca
      241 accataggat gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga
      301 agcgatgatt tttgatctat taacagatat ataaatgcaa aaactgcata accactttaa
      361 ctaatacttt caacattttc ggtttgtatt acttcttatt caaatgtaat aaaagtatca
      421 acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaacc ccggattctg
      481 aagcggccgc actagtggat cccccgggct gcaggaattc gatatcaagc ttatcgatac
      541 cgtcgacaat gttgtctttc acaaccaaga atagtttccg cttattactt ttaatactgt
      601 catgcatatc gacgatacgc gcacaatttt tcgtgcaatc atcatcttcg aattcgggtc
      661 tttctaaaaa aaatagaaac atcgtcattg gttgtgtggt tggtattggt gtaccactga
      721 ttctggtgat actggcttta atttacatgt tttgtatcca atcatcaagg acggacgcgt
      781 cgacctcgac aatgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      841 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
      901 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
      961 ggccaacact tgtcactact ttcacttatg gtgttcaatg cttttcaaga tacccagatc
     1021 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
     1081 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
     1141 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
     1201 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
     1261 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
     1321 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     1381 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc
     1441 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     1501 acaaaggtgg atctggaggt tcaggtggaa gtcctaggtt ggctactaca cttgaacgta
     1561 ttgagaagaa ctttgtcatt actgacccaa gattgccaga taatcccatt atattcgcgt
     1621 ccgatagttt cttgcagttg acagaatata gccgtgaaga aattttggga agaaactgca
     1681 ggtttctaca aggtcctgaa actgatcgcg cgacagtgag aaaaattaga gatgccatag
     1741 ataaccaaac agaggtcact gttcagctga ttaattatac aaagagtggt aaaaagttct
     1801 ggaacctctt tcacttgcag cctatgcgag atcagaaggg agatgtccag tactttattg
     1861 gggttcagtt ggatggaact gagcatgtcc gagatgctgc cgagagagag gcagtcatgc
     1921 tgattaaaac tgcagaagag attcgggggt cgattgatac ctgggtgtaa ctcgagtcat
     1981 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
     2041 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
     2101 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc
     2161 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat
     2221 ttgcggccgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2281 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2341 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2401 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2461 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2521 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2581 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2641 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2701 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2761 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2821 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2881 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2941 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3001 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3061 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3121 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3181 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3241 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3301 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3361 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3421 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3481 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3541 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3601 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3661 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3721 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3781 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3841 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3901 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3961 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4021 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4081 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4141 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4201 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4261 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4321 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4381 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4441 aaaataggcg tatcacgagg ccaactgtgg gaatactcag gtatcgtaag atgcaagagt
     4501 tcgaatctct tagcaaccat tatttttttc ctcaacataa cgagaacaca caggggcgct
     4561 atcgcacaga atcaaattcg atgactggaa attttttgtt aatttcagag gtcgcctgac
     4621 gcatatacct ttttcaactg aaaaattggg agaaaaagga aaggtgagag gccggaaccg
     4681 gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca atacttgaag
     4741 ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca atcgtcttac
     4801 tttctaactt ttcttacctt ttacatttca gcaatatata tatatatttc aaggatatac
     4861 cattctaatg tctgccccta tgtctgcccc taagaagatc gtcgttttgc caggtgacca
     4921 cgttggtcaa gaaatcacag ccgaagccat taaggttctt aaagctattt ctgatgttcg
     4981 ttccaatgtc aagttcgatt tcgaaaatca tttaattggt ggtgctgcta tcgatgctac
     5041 aggtgtccca cttccagatg aggcgctgga agcctccaag aaggttgatg ccgttttgtt
     5101 aggtgctgtg ggtggtccta aatggggtac aggtagtgtt agacctgaac aaggtttact
     5161 aaaaatccgt aaagaacttc aattgtacgc caacttaaga ccatgtaact ttgcatccga
     5221 ctctctttta gacttatctc caatcaagcc acaatttgct aaaggtactg acttcgttgt
     5281 tgtcagagaa ttagtgggag gtatttactt tggtaagaga aaggaagacg atggtgatgg
     5341 tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg caaagaatca caagaatggc
     5401 cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg ataaagctaa
     5461 tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca agaacgaatt
     5521 tcctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc tagttaagaa
     5581 cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata tcatctccga
     5641 tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct tggcctcttt
     5701 gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg ctccagattt
     5761 gccaaagaat aaggttgacc ctatcgccac tatcttgtct gctgcaatga tgttgaaatt
     5821 gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa aggttttgga
     5881 tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag tcggtgatgc
     5941 tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat tctctttttt tatgatattt
     6001 gtacataaac tttataaatg aaattcataa tagaaacgac acgaaattac aaaatggaat
     6061 atgttcatag ggtaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6121 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6181 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6241 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6301 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6361 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6421 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     6481 tgaattcgag ct
//
