LOCUS       Exported                6895 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  N-terminal 9myc-AID* degron cassette with CUP1 promoter and KanR 
            marker for S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    pKan-PCUP1-9myc-AID*(N)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6895)
  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 6895)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6895
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..235
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             297..326
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             333..362
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             369..398
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             417..446
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             453..482
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             489..518
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             537..566
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             573..602
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             609..638
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(894..912)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(933..949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(933..949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(946..968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    957..973
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(981..1011)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1026..1047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1164..1181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1335..1923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1415..1434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2094..2954)
                     /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     2717..2736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2955..3059)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3127..3145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3183..3205)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3305..3324
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3348..3563
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3564..4367
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4498..4953)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4635..4656)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4847..4866
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5079..5101
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5093..5110
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5094..5110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5117..5136
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5117..5135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            5270..6626
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        5270..5613
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5614..6423
                     /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(5681..5700)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6429..6626
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 ctagttagaa aaagacattt ttgctgtcag tcactgtcaa gagattcttt tgctggcatt
       61 tcttctagaa gcaaaaagag cgatgcgtct tttccgctga accgttccag caaaaaagac
      121 taccaacgca atatggattg tcagaatcat ataaaagaga agcaaataac tccttgtctt
      181 gtatcaattg cattataata tcttcttgtt agtgcaatat catatagaag tcatcgaaat
      241 agatattaag aaaaacaaac tgtacacgtg tcatgtccgg ttctgctgct agtggtgaac
      301 aaaagttgat ttctgaagaa gatttgaacg gtgaacaaaa gctaatctcc gaggaagact
      361 tgaacggtga acaaaaatta atctcagaag aagacttgaa cggatcctct agaggtgaac
      421 aaaagttgat ttctgaagaa gatttgaacg gtgaacaaaa gctaatctcc gaggaagact
      481 tgaacggtga acaaaaatta atctcagaag aagacttgaa cggatcctct agaggtgaac
      541 aaaagttgat ttctgaagaa gatttgaacg gtgaacaaaa gctaatctcc gaggaagact
      601 tgaacggtga acaaaaatta atctcagaag aagacttgaa cggatccact agagaattca
      661 tgcctaaaga tccagccaaa cctccggcca aggcacaagt tgtgggatgg ccaccggtga
      721 gatcataccg gaagaacgtg atggtttcct gccaaaaatc aagcggtggc ccggaggcgg
      781 cggcgttcgt gaaggtatca atgctcgagg aagaacttca aattgaatct aaggacttgg
      841 gagctggtgc aggcgctgga gcgggtgcca gagctcggta ccagcttttg ttccctttag
      901 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
      961 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1021 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1081 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1141 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1201 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1261 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1321 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1381 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1441 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1501 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1561 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1621 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1681 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1741 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1801 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1861 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1921 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1981 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2041 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2101 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2161 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2221 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2281 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2341 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2401 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2461 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2521 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2581 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2641 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2701 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2761 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2821 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2881 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2941 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3001 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3061 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3121 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3181 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3241 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3301 tatgcggcat cagagcagat tgtactgaga gtgcaccata ccaccttttc aattcatcat
     3361 ttttttttta ttcttttttt tgatttcggt ttccttgaaa tttttttgat tcggtaatct
     3421 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     3481 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     3541 cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     3601 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     3661 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     3721 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     3781 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa
     3841 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     3901 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     3961 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     4021 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     4081 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     4141 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
     4201 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg
     4261 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     4321 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     4381 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     4441 taccctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     4501 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4561 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4621 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4681 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4741 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4801 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4861 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4921 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     4981 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5041 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5101 gacggccagt gaattgtaat acgactcact atagggcgaa ttggagctcc accgcggtgg
     5161 cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt aattaaggcg
     5221 cgccagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
     5281 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
     5341 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
     5401 ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc
     5461 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
     5521 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
     5581 cagttctcac atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg
     5641 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     5701 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     5761 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     5821 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     5881 gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa
     5941 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     6001 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     6061 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     6121 ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat
     6181 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     6241 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     6301 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     6361 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     6421 taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt
     6481 tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc
     6541 tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc
     6601 gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagtgt
     6661 cgaaaacgag gcggccgctc tagaactagt gctagccatt accgacattt ggcgctctag
     6721 aactagtgct agccattacc gacatttggg cgctatacgt gcatatgttc atgtatgtat
     6781 ctgtatttaa aacacttttg tattattttt cctcatatat gtgtataggt ttatacggat
     6841 gatttaatta ttacttcacc accctttatt tcaggctgat atcttagcct tgtta
//
