LOCUS       sequence-451859-        5620 bp    DNA     circular SYN 15-APR-2025
DEFINITION  To integrate TID-FKBP-NLS fusion at yeast promoter; yeast BioID with
            KanMX6 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5620)
  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 5620)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pFa6-TID-FKBP-NLS-3HA-KanMX6
FEATURES             Location/Qualifiers
     source          1..5620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1069..1389
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1396..1416
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1417..1506
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      1533..1720
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1588..1612)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1795..3151
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1795..2138
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2139..2948
                     /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(2206..2225)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2954..3151
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3255..3274)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3256..3274)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3361..3378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3532..4120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3612..3631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4291..5082,5083..5151))
                     /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     4914..4933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5152..5256)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5324..5342
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5380..5402)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5502..5521
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5602..5620
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5602..5619
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gcaagagatc cgcctgtggc cacgatgaaa gataacacgg ttcccttaaa attgatagcc
      121 cttcttgcca atggtgagtt tcattccggg gaacaacttg gcgaaactct gggaatgagc
      181 agggcagcga ttaacaagca tatccaaacc ttgagagact ggggagtaga tgtgttcacc
      241 gtgcctggta aaggttattc tttgccggaa cctatccctc tattaaatgc gaaacagata
      301 ctaggacaac tggacggtgg tagtgttgcc gtattgccgg ttgttgactc aaccaatcaa
      361 tacttgctgg acagaattgg cgaacttaaa agcggtgatg cgtgtattgc agaatatcag
      421 caagcaggga gaggctcaag aggtaggaaa tggttttcac cttttggcgc caatctttac
      481 ttaagtatgt tctggcgttt gaagagaggg cccgcggcta ttggtttagg tcccgttatc
      541 gggatagtta tggctgaagc gctacgtaaa cttggggccg acaaggtcag agtgaagtgg
      601 cctaatgatc tatacttgca agatagaaag ctagcgggta tcctggtgga gctagcggga
      661 atcacagggg acgctgccca aatagtaatc ggtgctggta taaatgtcgc aatgcgtaga
      721 gtcgaggaat cagtggtgaa ccagggctgg atcacgttgc aagaggcggg cataaacctg
      781 gaccgtaaca cccttgcagc gaccctaata agagaactac gtgctgcatt ggagcttttt
      841 gagcaggagg gcctggctcc gtacctaccc aggtgggaga aattagacaa tttcataaac
      901 cgtccagtca aactaataat tggagataag gagattttcg gtatttctcg tgggatagac
      961 aagcagggag cgttgttatt agaacaggat ggtgtcatta aaccttggat gggcggtgaa
     1021 attagtcttc gtagtgccga aaaggctagc ggttctggtg gtgctagcgg agtgcaggtg
     1081 gaaaccatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg
     1141 cactacaccg ggatgcttga agatggaaag aaatttgatt cctcccggga cagaaacaag
     1201 ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga aggggttgcc
     1261 cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc ctatggtgcc
     1321 actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt ggagcttcta
     1381 aaactggaaa ctccaccaaa aaagaaaaga aaagtttacc catacgatgt tcctgactat
     1441 gcgggctatc cgtatgacgt cccggactat gcaggatctt atccatatga cgttccagat
     1501 tacgctgctt gaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
     1561 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     1621 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     1681 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
     1741 gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
     1801 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
     1861 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
     1921 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
     1981 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
     2041 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
     2101 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg
     2161 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     2221 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     2281 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     2341 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     2401 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
     2461 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     2521 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     2581 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     2641 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     2701 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2761 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2821 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2881 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     2941 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     3001 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     3061 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     3121 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     3181 agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     3241 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     3301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3361 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3421 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3961 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5281 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5341 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5401 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5461 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5521 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     5581 cttatgtatc atacacatac gatttaggtg acactataga
//
