LOCUS       Exported                5051 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  C-terminal AID*-GFP degron cassette with HygR marker for S. 
            cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    pHyg-AID*-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5051)
  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 5051)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5051
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             226..942
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             226..939
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(268..289)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(529..548)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     876..897
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        999..1342
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1343..2371
                     /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"
     primer_bind     complement(2395..2413)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      2425..2614
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2688..2707)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2689..2707)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2794..2811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2965..3553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3045..3064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3724..4584)
                     /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     4347..4366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4585..4689)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4757..4775
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4813..4835)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4935..4954
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5035..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5035..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 ggaaagtgag caagggcgag
      241 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      301 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      361 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      421 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      481 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      541 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      601 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      661 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      721 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      781 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      841 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      901 gccgccggga tcactctcgg catggacgag ctgtacaagt aaaaaggcgc gccagatctg
      961 tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc
     1021 tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt
     1081 agcccataca tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc
     1141 gaagcaaaaa ttacggctcc tcgctgcgga cctgcgagca gggaaacgct cccctcacag
     1201 acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc
     1261 cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca
     1321 tcacatccga acataaacaa ccatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     1381 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     1441 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     1501 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     1561 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
     1621 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     1681 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
     1741 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     1801 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     1861 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     1921 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     1981 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     2041 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     2101 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga
     2161 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     2221 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     2281 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     2341 ccccagcact cgtccgaggg caaaggaata atctcgagtc atgtaattag ttatgtcacg
     2401 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     2461 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     2521 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     2581 aaccttgctt gagaaggttt tgggacgctc gaaggcttta acgagctcga attcatcgat
     2641 gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt
     2701 cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag
     2761 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2821 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2881 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2941 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3001 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3061 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3121 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     3181 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3241 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3301 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3361 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3421 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3481 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3541 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3601 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3661 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3721 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3781 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3841 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3901 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3961 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4021 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4081 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4141 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4201 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4261 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4321 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4381 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4441 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4501 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4561 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4621 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4681 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4741 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     4801 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4861 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4921 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
     4981 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     5041 gtgacactat a
//
