LOCUS       sequence-177899-        8434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression of mouse PKC Alpha with yeast PKC1 promoter and 
            ADH1 terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8434)
  AUTHORS   Soriano-Carot M, Quilis I, Bano MC, Igual JC
  TITLE     Protein kinase C controls activation of the DNA integrity 
            checkpoint.
  JOURNAL   Nucleic Acids Res. 2014 Jun;42(11):7084-95. doi: 10.1093/nar/gku373.
            Epub 2014 May 3.
  PUBMED    24792164
REFERENCE   2  (bases 1 to 8434)
  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: 
            "10.1093/nar/gku373"; journalName: "Nucleic Acids Res"; date: 
            "2014-06"; volume: "42"; issue: "11"; pages: "7084-95"
COMMENT     Sequence Label: isoform PKC Alpha (JCE599)
FEATURES             Location/Qualifiers
     source          1..8434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2678..2865
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2733..2757)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2925..2941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2925..2941)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2938..2960)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2949..2965
                     /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(join(2973..2979,2980..2997,2998..3003))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3018..3039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3156..3173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3327..3915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3407..3426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4086..4877,4878..4946))
                     /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     4709..4728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4947..5051)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5119..5137
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5162..5377
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5378..6181
                     /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"
     misc_feature    6516..7678
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      6516..7353
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     join(8420..8434,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8434,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagtggt tcagcttctg tttgagttgc aaccatggta
       61 tgcatgccgg tcacgctgaa gaatggtttg acagacataa tgtttgtccc actccaggtt
      121 gcacctgcca gtgtaataag tagtattgta caataaacca aatcttgcaa cttctcttta
      181 ttctcacaat atatatacat aaccaaagaa aaaacgtaat tagtataaca ttgtatttat
      241 gaggcgttgc tatcttcatt acccagcccg cctagtttta aaggcgaaaa aaaccgacga
      301 aaagagttaa caagcgtgga taataaagtt gatctactct atcgaaagta atacgacagg
      361 ttcaggagca gtaaaagagc tacgatttca ctgctattca atttggctga gtagagactg
      421 attattaacc attgtaaaag tctctgccac cactcgtcca cacttacgtt agaggtattc
      481 agcgtcgatc ccctttcagt atagattttt aaaactatat tagataatca ataggaataa
      541 aacctagaac agtagctgcg gagagaagct ctagcgagga aaagtaagta tagtatcaca
      601 catataggga gcagtttaca gtcggtacca tggctgacgt ttacccggcc aacgactcca
      661 cggcgtctca ggacgtggcc aaccgcttcg cccgcaaagg ggcgctgagg cagaagaacg
      721 tgcatgaggt gaaagaccac aaattcatcg cccgcttctt caagcaaccc accttctgca
      781 gccactgcac cgacttcatc tgggggtttg ggaaacaagg cttccagtgc caagtttgct
      841 gttttgtggt tcataagagg tgccatgagt tcgttacgtt ctcttgtccg ggtgcggata
      901 agggacctga cactgacgac cccaggagca agcacaagtt caaaatccac acatacggaa
      961 gccctacctt ctgtgatcac tgtgggtccc tgctctatgg acttatccac caagggatga
     1021 aatgtgacac ctgcgacatg aatgttcaca agcagtgtgt gatcaatgtc cctagcctct
     1081 gcggaatgga tcacacagag aagagggggc ggatttatct gaaggctgag gtcactgatg
     1141 aaaagctcca cgtcacggta cgagatgcaa aaaatctaat ccctatggat ccaaatgggc
     1201 tttcggatcc ttatgtgaag ctgaaactta tccctgaccc caagaatgag agcaaacaga
     1261 aaaccaaaac catccgctcc acactgaatc ctcagtggaa tgagtccttc acgttcaaat
     1321 taaaaccttc agacaaagac cggcgactgt ctgtagaaat ctgggactgg gatcggacga
     1381 ctcggaatga cttcatggga tccctttcct ttggtgtctc agagctaatg aagatgccgg
     1441 ccagtggatg gtataaactg ctcaaccaag aagagggcga atattacaat gtgcccattc
     1501 cagaaggaga tgaagaaggc aacatggaac tcaggcagaa gtttgagaaa gccaagctag
     1561 gccctgctgg taacaaagtc atcagccctt cagaagacag aaagcaacca tccaacaacc
     1621 tggacagagt gaaactcaca gacttcaact tcctcatggt gctggggaag gggagttttg
     1681 ggaaggtgat gcttgctgac aggaagggaa cggaggaact gtacgccatc aagatcctga
     1741 agaaggacgt ggtgatccag gacgacgacg tggagtgcac catggtggag aagcgcgtgc
     1801 tggccctgct ggacaagccg ccatttctga cacagctgca ctcctgcttc cagacagtgg
     1861 accggctgta cttcgtcatg gaatacgtca acggcgggga cctcatgtac cacattcagc
     1921 aagtcgggaa atttaaggag ccacaagcag tattctacgc agccgagatc tccatcggac
     1981 tgttcttcct tcataaaaga gggatcattt acagggatct gaagctggac aatgtcatgc
     2041 tggactcaga agggcacatc aaaatcgccg acttcgggat gtgcaaggaa cacatgatgg
     2101 atggagtcac gaccaggacc ttctgtggga ctccggacta cattgcccca gagataatcg
     2161 cttaccagcc gtacgggaag tctgtagatt ggtgggcgta cggtgtgctg ctgtacgaga
     2221 tgctagccgg gcagcctccg tttgatggtg aagatgaaga tgaactgttt cagtctataa
     2281 tggagcacaa cgtgtcctac cccaaatcct tgtccaagga agccgtctcc atctgcaaag
     2341 gacttatgac caaacaccct gccaagcggc tgggctgcgg gcccgaggga gagagggatg
     2401 tcagagagca tgccttcttc aggaggatcg actgggagaa actggagaac agggagatcc
     2461 aaccaccatt caagcccaaa gtgtgtggca aaggagcaga aaactttgac aagttcttca
     2521 cgcgaggaca gcctgtctta acaccaccag atcagctggt cattgctaac atagaccaat
     2581 ctgattttga agggttctcg tatgtcaacc cccagtttgt gcacccaatc ttgcaaagtg
     2641 cagtatgagt cgactagggc gcgccacttc taaataagcg aatttcttat gatttatgat
     2701 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     2761 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     2821 tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga
     2881 taacagggta atatctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
     2941 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3001 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3061 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3121 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3181 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3241 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3301 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3361 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3421 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3481 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3541 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3601 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3661 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3721 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3781 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3841 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3901 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3961 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4021 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4081 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4141 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4201 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4261 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4321 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4381 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4441 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4501 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4561 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4621 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4681 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4741 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4801 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4861 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4921 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4981 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5041 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5101 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattagct
     5161 tttcaattca tcattttttt tttattcttt tttttgattt cggtttcctt gaaatttttt
     5221 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     5281 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     5341 gcacagaaca aaaacatgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
     5401 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
     5461 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
     5521 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
     5581 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
     5641 cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
     5701 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
     5761 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     5821 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     5881 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     5941 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     6001 caagggagac gcattgggtc aacagtatag aaccgtggat gatgtggtct ctacaggatc
     6061 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     6121 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     6181 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     6241 attatatcag ttattaccca attctcatgt ttgacagctt atcatcgatc gtccaactgc
     6301 atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac
     6361 tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt
     6421 ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga
     6481 tttctgactg ggttggaagg caagagagcc ccgagagctt acattttatg ttagctggtg
     6541 gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg
     6601 taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca
     6661 aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt
     6721 gcctgcaagc cttttgaaaa gcaagcataa aagatctaaa cataaaatct gtaaaataac
     6781 aagatgtaaa gataatgcta aatcatttgg ctttttgatt gattgtacag gaaaatatac
     6841 atcgcagggg gttgactttt accatttcac cgcaatggaa tcaaacttgt tgaagagaat
     6901 gttcacaggc gcatacgcta caatgacccg attcttgcta gccttttctc ggtcttgcaa
     6961 acaaccgccg gcagcttagt atataaatac acatgtacat acctctctcc gtatcctcgt
     7021 aatcattttc ttgtatttat cgtcttttcg ctgtaaaaac tttatcacac ttatctcaaa
     7081 tacacttatt aaccgctttt actattatct tctacgctga cagtaatatc aaacagtgac
     7141 acatattaaa cacagtggtt tctttgcata aacaccatca gcctcaagtc gtcaagtaaa
     7201 gatttcgtgt tcatgcagat agataacaat ctatatgttg ataattagcg ttgcctcatc
     7261 aatgcgagat ccgtttaacc ggaccctagt gcacttaccc cacgttcggt ccactgtgtg
     7321 ccgaacatgc tccttcacta ttttaacatg tggaattaat tctcatgttt gacagcttat
     7381 catcgaactc taagaggtga tacttattta ctgtaaaact gtgacgataa aaccggaagg
     7441 aagaataaga aaactcgaac tgatctataa tgcctatttt ctgtaaagag tttaagctat
     7501 gaaagcctcg gcattttggc cgctcctagg tagtgctttt tttccaagga caaaacagtt
     7561 tctttttctt gagcaggttt tatgtttcgg taatcataaa caataaataa attatttcat
     7621 ttatgtttaa aaataaaaaa taaaaaagta ttttaaattt ttaaaaaagt tgattataag
     7681 catgtgacct tttgcaagca attaaatttt gcaatttgtg atttaggcaa aagttactat
     7741 ttctggctcg tgtaatatat gtatgctaat gtgaactttt acaaagtcga tatggactta
     7801 gtcaaaagaa attttcttaa aaatatatag cactagccaa tttagcactt ctttatgaga
     7861 tatattatag actttattaa gccagatttg tgtattatat gtatttaccc ggcgaatcat
     7921 ggacatacat tctgaaatag gtaatattct ctatggtgag acagcataga taacctagga
     7981 tacaagttaa aagctagtac tgttttgcag taattttttt cttttttata agaatgttac
     8041 cacctaaata agttataaag tcaatagtta agtttgatat ttgattgtaa aataccgtaa
     8101 tatatttgca tgatcaaaag gctcaatgtt gactagccag catgtcaacc actatattga
     8161 tcaccgatat taggacttcc acaccaacta gtaatatgac aataaattca agatattctt
     8221 catgagaatg gcccagcgat tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     8281 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     8341 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     8401 tgggtaacgc cagggttttc ccagtcacga cgtt
//
