LOCUS       sequence-451856-        5065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To insert biotin ligase, BASU, at the C-terminus of genes; yeast 
            BioID, KanMX6 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5065)
  AUTHORS   Pfannenstein J, Tyryshkin M, Gulden ME, Doud EH, Mosley AL, Reese JC
  TITLE     Characterization of BioID tagging systems in budding yeast and 
            exploring the interactome of the Ccr4-Not complex.
  JOURNAL   G3 (Bethesda). 2024 Sep 13:jkae221. doi: 10.1093/g3journal/jkae221.
  PUBMED    39271111
REFERENCE   2  (bases 1 to 5065)
  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: "G3 
            (Bethesda). 2024 Sep 13:jkae221. doi: 10.1093/g3journal/jkae221."
COMMENT     Sequence Label: pFA6a-BASU-HA3-KanMX6
FEATURES             Location/Qualifiers
     source          1..5065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             856..945
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      978..1165
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1033..1057)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1240..2596
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1240..1583
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1584..2393
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1651..1670)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2399..2596
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2700..2719)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2701..2719)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2806..2823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2977..3565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3057..3076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3736..4527,4528..4596))
                     /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     4359..4378
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4597..4701)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4769..4787
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4825..4847)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4947..4966
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5047..5065
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5047..5064
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 ggcaagctga gcgaaagtga gattagattc ggtttaaaaa ctgaggtcat gggtcaacat
      121 ttgatatacc atgacgtact gtctagcaca caaaagaccg ctcacgagtt agcaaacaat
      181 aacgctcccg aaggtacatt ggtcgttgcg gataaacaaa cggctggtag aggcggaatg
      241 agtagggtct ggcattcaca agaaggaaat ggtgtgtgga tgtccttgat actaaggcca
      301 gacataccgt tacaaaaaac cccgcaacta acccttttgg ctgcggtggc cgtggttcag
      361 gggatcgagg aagccgctgg gatccaaacg gacataaaat ggccgaatga cattctaatc
      421 aacggcaaaa aaacagtcgg gattctgacc gaaatgcaag ccgaagagga cagggtaaga
      481 agtgtgatca ttgggattgg catcaatgta aatcagcaac ctaatgattt cccggacgaa
      541 ttaaaggaca tagccacctc tcttagccaa gccgctggcg aaaaaataga ccgtgctgga
      601 gtgatccagc atatacttct atgctttgag aagaggtaca gagactacat gactcatggt
      661 tttacgccaa taaagctgtt gtgggaatcc tacgctctgg gaataggaac taacatgcgt
      721 gctagaacac tgaatggtac attttacggt aaggccctag ggattgacga cgaaggtgta
      781 cttcttcttg agacaaacga gggcataaaa aagatataca gtgctgacat atctttgagg
      841 ttaattaaca tcttttaccc atacgatgtt cctgactatg cgggctatcc ctatgacgtc
      901 ccggactatg caggatccta tccatatgac gttccagatt acgctgctca gtgctgaggc
      961 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     1021 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     1081 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     1141 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
     1201 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
     1261 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
     1321 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
     1381 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
     1441 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
     1501 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
     1561 atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat
     1621 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1681 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     1741 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1801 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1861 ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg
     1921 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     1981 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2041 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2101 ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca
     2161 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2221 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2281 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2341 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac
     2401 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     2461 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     2521 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     2581 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc
     2641 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     2701 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     2761 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2821 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2881 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2941 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3001 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3061 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3121 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3181 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3241 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3301 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3361 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3421 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3481 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3601 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3661 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3721 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3781 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3841 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3901 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3961 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4021 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4081 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4141 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4201 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4261 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4321 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4381 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4441 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4501 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4561 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4621 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4681 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4741 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4801 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4861 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4921 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4981 tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca
     5041 catacgattt aggtgacact ataga
//
