LOCUS       sequence-191042-        4336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal AID* degron cassette with HygR marker for S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4336)
  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 4336)
  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: "10.1002/yea.2967"; 
            journalName: "Yeast"; date: "2013-09"; volume: "30"; issue: "9"; 
            pages: "341-51"
COMMENT     Sequence Label: pHyg-AID*
FEATURES             Location/Qualifiers
     source          1..4336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        301..644
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             645..1673
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             645..1673
                     /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      1681..1928
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1697..1715)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1990..2009)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1991..2009)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2096..2113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2267..2855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2347..2366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3026..3817,3818..3886))
                     /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     3649..3668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3887..3991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4059..4077
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4115..4137)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4237..4256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg
       61 gtgcaggcgc cggggctggt gctggggcag gtacccctaa agatccagcc aaacctccgg
      121 ccaaggcaca agttgtggga tggccaccgc ttgatttttg gcaggaaacc atcacgttct
      181 tccggtatga tctcaccggt ggcccggagg cggcggcgtt cgtgaagtga tcacccgggt
      241 taattaaggc gcgccagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
      301 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
      361 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      421 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgcg gacctgcgag
      481 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      541 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      601 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaaaagcctg
      661 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
      721 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
      781 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
      841 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
      901 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
      961 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     1021 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     1081 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     1141 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     1201 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     1261 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     1321 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     1381 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     1441 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     1501 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     1561 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     1621 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taatctcgag
     1681 tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg
     1741 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt
     1801 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt
     1861 acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt
     1921 taacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc
     1981 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat
     2041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3001 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3061 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4021 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4081 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4141 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4201 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4261 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     4321 gtatcataca catacg
//
