LOCUS       sequence-177666-        8629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression of mouse PKC Epsilon with yeast PKC1 promoter and 
            ADH1 terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8629)
  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 8629)
  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 Epsilon (JCE592)
FEATURES             Location/Qualifiers
     source          1..8629
                     /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      2873..3060
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2928..2952)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3120..3136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3120..3136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3133..3155)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3144..3160
                     /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(3168..3174,3175..3192,3193..3198))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3213..3234
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3351..3368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3522..4110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3602..3621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4281..5072,5073..5141))
                     /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     4904..4923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5142..5246)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5314..5332
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5357..5572
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5573..6376
                     /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    6711..7873
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      6711..7548
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     join(8615..8629,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8629,1..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 tggtagtgtt caatggcctt cttaagatca
      661 aaatctgcga ggcggtgagc ttgaagccca cagcctggtc gctgcgccat gcggtgggac
      721 cccggccaca gacgttcctt ttggacccct acattgccct taacgtggac gactcgcgca
      781 tcggccaaac agccaccaag caaaagacca acagcccggc ctggcacgat gagttcgtca
      841 ccgatgtgtg caatgggcgc aagatcgagc tggctgtctt tcacgacgct cctatcggct
      901 acgacgactt cgtggccaac tgcaccatcc agttcgagga gctgctgcag aatgggagcc
      961 gtcacttcga ggactggatt gacctggagc cagaaggaaa agtgtacgtg atcatcgatc
     1021 tctcgggatc atcgggtgaa gcccctaaag acaatgaaga acgagtgttc agggagcgta
     1081 tgcggccaag gaagcggcaa ggggctgtca ggcgcagggt ccaccaggtc aatggccaca
     1141 agttcatggc cacctacttg cggcaaccca cctactgctc ccactgcaga gatttcatct
     1201 ggggtgtcat aggaaaacag ggatatcaat gtcaagtttg cacttgcgtt gtccacaagc
     1261 gatgtcatga gctcattatt acaaagtgcg ctgggctgaa gaaacaggaa acccctgacg
     1321 aggtgggctc ccaacggttc agcgtcaaca tgccccacaa gttcgggatc cacaactaca
     1381 aggtccccac gttctgtgac cactgtgggt ccctgctctg gggcctcttg cggcagggct
     1441 tgcagtgtaa agtctgcaaa atgaatgttc accggcgatg tgagaccaat gtggctccca
     1501 actgtggggt agacgccaga ggaattgcca aagtgctggc tgaccttggt gttactccag
     1561 acaaaatcac caacagtggc caaaggagga aaaagctcgc tgctggtgct gagtccccac
     1621 agccggcttc tggaaactcc ccatctgaag acgaccgatc caagtcagcg cccacctccc
     1681 cttgtgacca ggaactaaaa gaacttgaaa acaacattcg gaaggccttg tcatttgaca
     1741 accgaggaga ggagcaccga gcgtcgtcgg ccaccgatgg ccagctggca agccccggag
     1801 agaacgggga agtccggcca ggccaggcca agcgcttggg gctggatgag ttcaacttca
     1861 tcaaggtgtt gggcaaaggc agctttggca aggtcatgtt ggcggaactc aaaggcaaag
     1921 atgaagtcta cgctgtgaag gtcttgaaga aggacgttat cctacaagac gatgatgtgg
     1981 actgcacaat gacagagaag aggattttgg ctctggctcg gaaacaccct tatctaaccc
     2041 aactctattg ctgcttccag accaaggacc gcctcttctt cgtcatggaa tatgtaaatg
     2101 gtggagacct catgttccag attcagcggt cccgaaaatt tgatgagcct cgttctcggt
     2161 tctatgccgc agaggtcaca tcagccctca tgtttctcca ccagcacgga gtgatctaca
     2221 gggatttgaa actggacaac atccttctag atgcagaagg ccactgcaag ctggctgact
     2281 ttgggatgtg caaggaaggg attatgaatg gtgtgacaac taccaccttc tgtgggactc
     2341 ctgactacat agctccagag atcctacagg agttggagta cggcccctca gtggactggt
     2401 gggccctggg ggtgctgatg tacgagatga tggctgggca gccccccttt gaagctgaca
     2461 acgaggacga cttgttcgaa tccatccttc atgatgatgt tctctatcct gtctggctta
     2521 gcaaggaagc tgtcagcatc ctgaaagctt tcatgaccaa gaacccgcac aagcgcctgg
     2581 gctgtgtggc agcgcagaac ggggaggacg ccatcaagca acatccattc ttcaaggaga
     2641 ttgactgggt actgctggag cagaagaaaa tcaagccccc cttcaagccg agaattaaaa
     2701 ccaaaagaga tgtcaataac tttgaccaag actttacgcg ggaagagcca atacttacac
     2761 ttgtggatga agcaatcatt aagcagatca accaggaaga attcaaaggc ttctcctact
     2821 ttggtgaaga cctgatgccc tgagtcgact agggcgcgcc acttctaaat aagcgaattt
     2881 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     2941 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     3001 ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg
     3061 cagatccgct agggataaca gggtaatatc tgcaggcatg caagcttggc gtaatcatgg
     3121 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3181 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3241 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3301 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3961 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5221 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5281 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5341 cttcaagaat tagcttttca attcatcatt ttttttttat tctttttttt gatttcggtt
     5401 tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     5461 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     5521 attcttaacc caactgcaca gaacaaaaac atgcaggaaa cgaagataaa tcatgtcgaa
     5581 agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa
     5641 tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga
     5701 attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga
     5761 tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa
     5821 gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata cagtcaaatt
     5881 gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg
     5941 tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga
     6001 acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga
     6061 atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat
     6121 tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg
     6181 tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg tggatgatgt
     6241 ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga
     6301 tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg
     6361 cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat
     6421 tagagcttca atttaattat atcagttatt acccaattct catgtttgac agcttatcat
     6481 cgatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc
     6541 cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt gcagcttcac
     6601 agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca ggtgaacttt
     6661 tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgag agcttacatt
     6721 ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg
     6781 ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa
     6841 tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt
     6901 attgtttgtg cacttgcctg caagcctttt gaaaagcaag cataaaagat ctaaacataa
     6961 aatctgtaaa ataacaagat gtaaagataa tgctaaatca tttggctttt tgattgattg
     7021 tacaggaaaa tatacatcgc agggggttga cttttaccat ttcaccgcaa tggaatcaaa
     7081 cttgttgaag agaatgttca caggcgcata cgctacaatg acccgattct tgctagcctt
     7141 ttctcggtct tgcaaacaac cgccggcagc ttagtatata aatacacatg tacatacctc
     7201 tctccgtatc ctcgtaatca ttttcttgta tttatcgtct tttcgctgta aaaactttat
     7261 cacacttatc tcaaatacac ttattaaccg cttttactat tatcttctac gctgacagta
     7321 atatcaaaca gtgacacata ttaaacacag tggtttcttt gcataaacac catcagcctc
     7381 aagtcgtcaa gtaaagattt cgtgttcatg cagatagata acaatctata tgttgataat
     7441 tagcgttgcc tcatcaatgc gagatccgtt taaccggacc ctagtgcact taccccacgt
     7501 tcggtccact gtgtgccgaa catgctcctt cactatttta acatgtggaa ttaattctca
     7561 tgtttgacag cttatcatcg aactctaaga ggtgatactt atttactgta aaactgtgac
     7621 gataaaaccg gaaggaagaa taagaaaact cgaactgatc tataatgcct attttctgta
     7681 aagagtttaa gctatgaaag cctcggcatt ttggccgctc ctaggtagtg ctttttttcc
     7741 aaggacaaaa cagtttcttt ttcttgagca ggttttatgt ttcggtaatc ataaacaata
     7801 aataaattat ttcatttatg tttaaaaata aaaaataaaa aagtatttta aatttttaaa
     7861 aaagttgatt ataagcatgt gaccttttgc aagcaattaa attttgcaat ttgtgattta
     7921 ggcaaaagtt actatttctg gctcgtgtaa tatatgtatg ctaatgtgaa cttttacaaa
     7981 gtcgatatgg acttagtcaa aagaaatttt cttaaaaata tatagcacta gccaatttag
     8041 cacttcttta tgagatatat tatagacttt attaagccag atttgtgtat tatatgtatt
     8101 tacccggcga atcatggaca tacattctga aataggtaat attctctatg gtgagacagc
     8161 atagataacc taggatacaa gttaaaagct agtactgttt tgcagtaatt tttttctttt
     8221 ttataagaat gttaccacct aaataagtta taaagtcaat agttaagttt gatatttgat
     8281 tgtaaaatac cgtaatatat ttgcatgatc aaaaggctca atgttgacta gccagcatgt
     8341 caaccactat attgatcacc gatattagga cttccacacc aactagtaat atgacaataa
     8401 attcaagata ttcttcatga gaatggccca gcgattatgc ggtgtgaaat accgcacaga
     8461 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     8521 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     8581 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
