LOCUS       sequence-191049-        4523 bp    DNA     circular SYN 14-APR-2025
DEFINITION  C-terminal AID*-6HA degron cassette with HygR marker for S. 
            cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4523)
  AUTHORS   Morawska M, Ulrich HD
  TITLE     An expanded tool kit for the auxin-inducible degron system in 
            budding yeast.
  JOURNAL   Yeast. 2013 Sep;30(9):341-51. doi: 10.1002/yea.2967. Epub 2013 Jul 
            23.
  PUBMED    23836714
REFERENCE   2  (bases 1 to 4523)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/yea.2967"; 
            journalName: "Yeast"; date: "2013-09"; volume: "30"; issue: "9"; 
            pages: "341-51"
COMMENT     Sequence Label: pHyg-AID*-6HA
FEATURES             Location/Qualifiers
     source          1..4523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             226..315
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             322..411
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     promoter        471..814
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             815..1843
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             815..1843
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1851..2098
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1867..1885)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2160..2179)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2161..2179)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2266..2283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2437..3025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2517..2536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3196..3987,3988..4056))
                     /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     3819..3838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4057..4161)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4229..4247
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4285..4307)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4407..4426
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4507..4523,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4507..4523,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggtgcagg cgccggggct
       61 ggtgctgggg caggtacccc taaagatcca gccaaacctc cggccaaggc acaagttgtg
      121 ggatggccac cggtgagatc ataccggaag aacgtgatgg tttcctgcca aaaatcaagc
      181 ggtggcccgg aggcggcggc gttcgtgaag gtatcacccg ggaaataccc atacgatgtt
      241 cctgactatg cgggctatcc ctatgacgtc ccggactatg caggatccta tccatatgac
      301 gttccagatt acgcttctag atacccatac gatgttcctg actatgcggg ctatccctat
      361 gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc ttaaaaaggc
      421 gcgccagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg
      481 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
      541 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
      601 gccgcacggc gcgaagcaaa aattacggct cctcgctgcg gacctgcgag cagggaaacg
      661 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
      721 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
      781 acagttctca catcacatcc gaacataaac aaccatgggt aaaaagcctg aactcaccgc
      841 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
      901 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
      961 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     1021 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
     1081 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     1141 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     1201 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     1261 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     1321 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     1381 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     1441 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     1501 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     1561 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     1621 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     1681 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     1741 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     1801 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taatctcgag tcatgtaatt
     1861 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     1921 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     1981 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     2041 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taacgagctc
     2101 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     2161 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     2221 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2281 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2341 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2941 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3001 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3061 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3121 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3181 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3481 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3781 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4201 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4261 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4321 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4381 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4441 tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca
     4501 catacgattt aggtgacact ata
//
