LOCUS       sequence-209183-        4773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tag protein of interest with PIF6 in Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4773)
  AUTHORS   Yang X, Jost AP, Weiner OD, Tang C
  TITLE     A light-inducible organelle-targeting system for dynamically 
            activating and inactivating signaling in budding yeast.
  JOURNAL   Mol Biol Cell. 2013 Aug;24(15):2419-30. doi: 
            10.1091/mbc.E13-03-0126. Epub 2013 Jun 12.
  PUBMED    23761071
REFERENCE   2  (bases 1 to 4773)
  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: 
            "10.1091/mbc.E13-03-0126"; journalName: "Mol Biol Cell"; date: 
            "2013-08"; volume: "24"; issue: "15"; pages: "2419-30"
COMMENT     Sequence Label: linker-mCitrine-PIF6-NAT
FEATURES             Location/Qualifiers
     source          1..4773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            115..1234
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        115..458
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             459..1031
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1037..1234
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1639..2352)
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSRLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2410..2429)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2411..2429)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2516..2533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2687..3275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2767..2786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3446..4237,4238..4306))
                     /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     4069..4088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4307..4411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4479..4497
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4535..4557)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4657..4676
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4757..4773,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4757..4773,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
      121 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      181 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      241 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
      301 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
      361 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
      421 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaccact cttgacgaca
      481 cggcttaccg gtaccgcacc agtgtcccgg gggacgccga ggccatcgag gcactggatg
      541 ggtccttcac caccgacacc gtcttccgcg tcaccgccac cggggacggc ttcaccctgc
      601 gggaggtgcc ggtggacccg cccctgacca aggtgttccc cgacgacgaa tcggacgacg
      661 aatcggacga cggggaggac ggcgacccgg actcccggac gttcgtcgcg tacggggacg
      721 acggcgacct ggcgggcttc gtggtcgtct cgtactccgg ctggaaccgc cggctgaccg
      781 tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg ggtcgggcgc gcgttgatgg
      841 ggctcgcgac ggagttcgcc cgcgagcggg gcgccgggca cctctggctg gaggtcacca
      901 acgtcaacgc accggcgatc cacgcgtacc ggcggatggg gttcaccctc tgcggcctgg
      961 acaccgccct gtacgacggc accgcctcgg acggcgagca ggcgctctac atgagcatgc
     1021 cctgccccta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     1081 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     1141 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     1201 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
     1261 tccagtgtcg aaaacgagct cgaattctta gtcaacatgt ttattgcttt ccaacatgtt
     1321 tgtttcatgg gcagcagcaa catgactttg tggaacaacc tgcgtgtccc cgagatttgt
     1381 gatggcacca ccaccaccat ggatgatact cttcatgaaa tcctcgttat actctgcctc
     1441 atacaaatcc atgatcgatt tggtacgttg attatgcaga gaaacgttgg atctttggcc
     1501 ctttgcaaga atctggccat tctcaaacac aagctccata tactcttgat cgcttaacct
     1561 gcaacaataa tcggttggta agaacatcat atcgctactc cctgagccgg atccgctccc
     1621 tgaacctagg gaactacctt tgtacaattc atccatacca tgggtaatac cagcagcagt
     1681 aacaaattct aacaagacca tgtggtctct cttttcgttt ggatctttgg ataatctaga
     1741 ttgataggat aagtaatggt tgtctggtaa caagactgga ccatcaccaa ttggagtatt
     1801 ttgttgataa tggtcagcta attgaacaga accatcttca atgttgtgtc taattttgaa
     1861 gttaactttg ataccattct tttgtttgtc agccatgatg taaacattgt gagagttata
     1921 gttgtattcc aatttgtgac ctaaaatgtt accatcttct ttaaaatcaa taccttttaa
     1981 ttcgattcta ttaactaagg tatcaccttc aaacttgact tcagctctgg tcttgtagtt
     2041 accgtcatct ttgaaaaaaa tagttctttc ttgaacataa ccttctggca tggcagactt
     2101 gaaaaagtca tgttgtttca tatgatctgg gtatctagca aaacacatca aaccataacc
     2161 taaagtagtg actaaggttg gccatggaac tggcaattta ccagtagtac aaataaattt
     2221 taaggtcaat ttaccgtaag tagcatcacc ttcaccttca ccggagacag aaaatttgtg
     2281 accattaaca tcaccatcta attcaaccaa aattgggaca acaccagtga ataattcttc
     2341 acctttagac attgctttgt taattaaacc agcaccgtca cctgcggcgg cccgcggatc
     2401 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat
     2461 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3121 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3241 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3301 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3361 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3421 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3481 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3541 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3601 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3661 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3721 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3781 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3841 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3901 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3961 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4021 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4081 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4141 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4201 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4261 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4321 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4381 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4441 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4501 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4561 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4621 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4681 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     4741 gtatcataca catacgattt aggtgacact ata
//
