LOCUS       sequence-244952-        4990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For C-terminal tagging with TurboID in yeast including 
            nourseothricin-resistant marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4990)
  AUTHORS   Larochelle M, Bergeron D, Arcand B, Bachand F
  TITLE     Proximity-dependent biotinylation by TurboID to identify 
            protein-protein interaction networks in yeast.
  JOURNAL   J Cell Sci. 2019 May 7. pii: jcs.232249. doi: 10.1242/jcs.232249.
  PUBMED    31064814
REFERENCE   2  (bases 1 to 4990)
  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: "J Cell Sci.
            2019 May 7. pii: jcs.232249. doi: 10.1242/jcs.232249."
COMMENT     Sequence Label: pFA6a-TurboID-3MYC-natMX6 (pFB1434)
FEATURES             Location/Qualifiers
     source          1..4990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1027..1116
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     terminator      1140..1327
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1195..1219)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1402..2521
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1402..1745
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1746..2318
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2324..2521
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2625..2644)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2626..2644)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2731..2748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2902..3490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2982..3001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3661..4452,4453..4521))
                     /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     4284..4303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4522..4626)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4694..4712
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4750..4772)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4872..4891
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4972..4990
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4972..4989
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 atctttaaag acaatacagt gccgctcaaa ttgatagccc tccttgcaaa tggagagttt
      121 cacagcggtg aacaactggg cgagactctg ggtatgtcac gcgcagctat aaataagcat
      181 atacagacac tcagggattg gggggttgac gtatttacag tgccaggcaa gggctattct
      241 ctcccagagc ccataccact tttgaatgct aaacagatac ttggtcaatt ggatggaggt
      301 agcgttgcgg tgctcccggt tgttgattcc acaaatcagt atctgctgga taggattgga
      361 gaactcaaaa gcggggatgc ctgtatcgcg gaataccaac aagctggtcg aggctcacga
      421 ggcagaaaat ggttctcccc gttcggggca aatctgtacc ttagcatgtt ttggcgcttg
      481 aaaagaggac ccgcggcgat aggactcggc cccgttattg gaattgttat ggccgaagct
      541 cttcgaaaat tgggtgctga caaggttcgg gtgaagtggc ctaacgattt gtatttgcag
      601 gaccggaaac tcgcaggaat tttggtcgag ctggcaggta ttactggtga cgcggctcaa
      661 attgttatag gtgccggcat caacgtcgct atgagacggg tcgaagagtc cgttgtaaat
      721 caagggtgga ttacactcca ggaggcaggc ataaatctcg accgaaatac gttggctgct
      781 accctgattc gagagcttag agctgcactc gaattgtttg aacaagaagg tctcgcacca
      841 tatcttccga gatgggagaa actcgataat ttcattaacc ggcctgtcaa gttgatcatt
      901 ggagataaag agattttcgg gatttccaga ggtattgaca aacaaggcgc tctcctcctt
      961 gagcaagatg gggtgatcaa gccttggatg gggggcgaga tctccctgag gtcagcagag
     1021 aaagacgagc agaaactcat ctctgaagaa gatctggaac aaaagttgat ttcagaagaa
     1081 gatctggaac agaagctcat ctctgaggaa gatctgtgag gcgcgccact tctaaataag
     1141 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     1201 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     1261 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
     1321 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
     1381 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
     1441 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
     1501 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
     1561 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
     1621 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
     1681 catatacttc ctttttaaat cttgctagga tacagttctc acatcacatc cgaacataaa
     1741 caaccatggg taccactctt gacgacacgg cttaccggta ccgcaccagt gtcccggggg
     1801 acgccgaggc catcgaggca ctggatgggt ccttcaccac cgacaccgtc ttccgcgtca
     1861 ccgccaccgg ggacggcttc accctgcggg aggtgccggt ggacccgccc ctgaccaagg
     1921 tgttccccga cgacgaatcg gacgacgaat cggacgacgg ggaggacggc gacccggact
     1981 ctcggacgtt cgtcgcgtac ggggacgacg gcgacctggc gggcttcgtg gtcgtctcgt
     2041 actccggctg gaaccgccgg ctgaccgtcg aggacatcga ggtcgccccg gagcaccggg
     2101 ggcacggggt cgggcgcgcg ttgatggggc tcgcgacgga gttcgcccgc gagcggggtg
     2161 ccgggcacct ctggctggag gtcaccaacg tcaacgcacc ggcgatccac gcgtaccggc
     2221 ggatggggtt caccctctgc ggcctggaca ccgccctgta cgacggcacc gcctcggacg
     2281 gcgagcaggc gctctacatg agcatgccct gcccctaatc agtactgaca ataaaaagat
     2341 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     2401 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     2461 gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact
     2521 gctgtcgatt cgatactaac gccgccatcc agtttaaacg agctcgaatt catcgatgat
     2581 atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt
     2641 attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg
     2701 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2761 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2821 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3301 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3421 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3481 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3541 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3601 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3661 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3721 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3781 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3841 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3901 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3961 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4021 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4081 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4141 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4201 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4261 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4321 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4381 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4441 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4501 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4561 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4621 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4681 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4741 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4801 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4861 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc
     4921 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     4981 acactataga
//
