LOCUS       sequence-176918-        9783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression of PKC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9783)
  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 9783)
  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: pPKC1 (JCE575)
FEATURES             Location/Qualifiers
     source          1..9783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4274..4290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4274..4290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4287..4309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4298..4314
                     /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(4322..4328,4329..4346,4347..4352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4367..4388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4505..4522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4676..5264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4756..4775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5435..6226,6227..6295))
                     /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     6058..6077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6296..6400)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6468..6486
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6511..6726
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             6727..7530
                     /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    7865..9027
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      7865..8702
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     join(9769..9783,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(9783,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacccgggg atcctctaga gtcgacctgc
       61 aggcatgccg gtcacgctga agaatggttt gacagacata atgtttgtcc cactccaggt
      121 tgcacctgcc agtgtaataa gtagtattgt acaataaacc aaatcttgca acttctcttt
      181 attctcacaa tatatataca taaccaaaga aaaaacgtaa ttagtataac attgtattta
      241 tgaggcattg ctatcttcat tacccagccc gcctagtttt aaaggcgaaa aaaaccgacg
      301 aaaagagtta acaagcgtgg ataataaagt tgatctactc tatcgaaagt aatacgacag
      361 gttcaggagc agtaaaagag ctacgatttc actgctattc aatttggctg agtagagact
      421 gattattaac cattgtaaaa gtctctgcca ccactcgtcc acacttacgt tagaggtatt
      481 cagcgtcgat cccctttcag tatagatttt taaaactata ttagataatc aataggaata
      541 aaacctagaa cagtagctgc ggagagaagc tctagcgagg aaaagtaagt atagtatcac
      601 acatataggg agcagtttac agtcatgagt ttttcacaat tggagcagaa cattaaaaaa
      661 aagatagccg tcgaagaaaa tattattcgg ggagcttctg ccctcaagaa aaagactagc
      721 aatgttatgg tcattcagaa atgtaatacg aatattagag aggcacgtca aaatcttgag
      781 tacttggagg atagcttaaa aaaattacgg ttgaagaccg ctcaacaaag tcagggtgaa
      841 aatggcagcg aagataatga gcgatgcaat tcgaaggaat acgggtttct ttctaccaaa
      901 tcgccaaatg aacacatatt ttctcgtttg gatttagtaa aatatgattg cccttctttg
      961 gcgcaaagaa tccaatacat gttgcaacaa ttagaattca aattacaggt ggaaaagcaa
     1021 taccaagaag ccaatactaa attgactaaa ttgtatcaaa ttgacggtga tcaacgtagt
     1081 agttctgcgg cagaaggggg tgctatggaa tccaaataca ggattcaaat gcttaacaaa
     1141 gctttgaaga aataccaagc tattaacgtt gactttgatc aatttaaaca ccaacccaat
     1201 gacataatgg ataatcaaca accgaaattc cgaagaaaac aactgactgg cgttctaaca
     1261 attggaataa ctgctgctag agatgttgac catatacaat caccgatgtt tgccaggaag
     1321 ccagagagct acgttactat aaaaatcgat gatacgatca aagccagaac gaagccttct
     1381 agaaatgaca ggtggagtga agattttcaa attccagttg aaaaagggaa tgaaatcgag
     1441 attactgtat atgataaggt gaacgactcg ctcattccag tggctataat gtggttactg
     1501 ctttccgata tcgctgaaga aattcgtaag aagaaagccg gacaaacaaa tgaacaacaa
     1561 ggatgggtca acgcttcaaa cattaatggt ggttcttctc tcgctagcga agagggaagc
     1621 acattgactt ctacatactc aaactcagcc attcagtcta cttcagcaaa gaatgtgcaa
     1681 ggcgaaaata catcaactag tcaaatcagt acgaattcat ggtttgtctt agaaccatcc
     1741 gggcagattt tgttaacact aggattccac aagtcttctc aaatagaaag gaagcagttg
     1801 atgggtggac tacatcgtca tggtgctatt atcaatagga aagaagagat attcgaacag
     1861 catggccacc actttgtaca aaaatcattt tacaacatta tgtgctgtgc atattgtggt
     1921 gatttcctcc ggtatactgg attccaatgt caagattgta aatttttatg tcacaaaaaa
     1981 tgttacacca atgttgttac taagtgtatt gctaaaactt ctactgatac ggatccggac
     2041 gaggcaaagt tgaaccaccg tattcctcat agattcctgc ctacttcaaa ccgtggtact
     2101 aaatggtgct gtcactgtgg ttatatttta ccatggggta gacataaagt acgcaaatgt
     2161 tctgaatgtg gtataatgtg tcatgctcaa tgtgctcatt tggttcccga tttctgtggc
     2221 atgtcgatgg aaatggctaa taaaattctg aaaacgattc aagacacaaa acgtaatcaa
     2281 gagaaaaaga agaggacagt tccgtcggca cagttggggt catctattgg tactgccaat
     2341 ggtagtgatc tttccccatc caaattagct gaaagggcaa acgctccttt gcctcctcag
     2401 ccaagaaaac atgataaaac tccttcgcct cagaaggtag gaagagattc tccaactaag
     2461 cagcatgatc ctattattga taaaagaatt tcactacaaa cgcacgggcg tgagaaacta
     2521 aataaattta tcgatgaaaa tgaggcctat ttaaatttta cagaaggtgc acaacaaacc
     2581 gccgaatttt catcaccaga gaaaacactg gatccaacgt cgaatagacg ttccttggga
     2641 ctcactgatt tatctattga gcatagtcaa acttgggaat ctaaagatga tttaatgaga
     2701 gatgaacttg aactgtggaa agcacaacgt gaggaaatgg aacttgaaat aaaacaagat
     2761 agtggggaaa tccaagagga tctagaagtt gatcacattg atctagagac aaaacagaaa
     2821 ctagactggg agaacaaaaa tgatttccgt gaagctgatt tgacaataga ttccacacac
     2881 acaaatccat tccgcgatat gaatagtgaa acctttcaaa ttgaacaaga ccatgcttca
     2941 aaggaagtac tacaagaaac agtgtccttg gctcctacaa gtactcatgc ctccagaacc
     3001 actgatcaac aatctccgca gaaatcacaa accagcacaa gcgcaaagca taaaaagaga
     3061 gcggctaaac gtcgtaaggt ttcattagat aatttcgtat tactcaaagt tcttggtaaa
     3121 ggtaattttg gtaaagttat tttatctaaa tcaaagaata ctgacaggct ttgtgccata
     3181 aaagttctga agaaagataa tattattcaa aatcatgacc tcgagagtgc aagagcagaa
     3241 aagaaagtat ttttgttagc cacaaaaact aaacacccat tcttaaccaa tctatactgc
     3301 tcttttcaaa ctgaaaaccg tatatatttt gctatggagt ttattggggg cggtgactta
     3361 atgtggcatg ttcaaaacca aagactatct gtaagaaggg ccaaatttta tgccgccgag
     3421 gtcttactgg ctctgaaata ttttcatgat aatggtgtaa tataccgtga tttgaagttg
     3481 gaaaacattc tattaactcc agaaggtcat ataaaaattg ccgattatgg tttgtgtaaa
     3541 gatgaaatgt ggtatggtaa tagaacttct acattttgtg gcacaccaga atttatggct
     3601 ccagaaattt taaaagagca agaatatacc aaagctgtcg attggtgggc atttggggtg
     3661 ctgctatatc aaatgctact gtgccaatct ccattctcag gagatgacga agatgaagtt
     3721 tttaacgcta tccttaccga tgaaccctta tacccaatag atatggcagg tgagattgta
     3781 caaatattcc aaggcctatt aacaaaagat cctgaaaaga ggttgggtgc tggtcccagg
     3841 gacgcagacg aagttatgga agaacctttc ttccgtaaca tcaactttga cgatatctta
     3901 aaccttcgcg tgaaaccacc ctacatccca gaaattaaat ctccggaaga tacatcatat
     3961 ttcgagcaag aattcacctc tgcgccaccc acacttacac ctctgccctc tgttttgaca
     4021 acgagccagc aagaagagtt tagaggattt tcctttatgc cagatgattt ggatttatga
     4081 agaaaaggtc atgccatgat ttatttaatt ttatataaaa catctaagcg gaaaagttga
     4141 actgttccgc ttcataggag gagagggttt ttttaatgag attttgttac ataaactgaa
     4201 actatccatt ttgatgcaat gcctcttttt tgttcgtacc tatcttggag catgcaagct
     4261 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4321 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4381 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4441 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4501 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4561 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4621 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4681 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4741 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4801 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4861 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4921 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4981 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5041 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5101 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5161 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5221 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5281 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5341 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5401 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5461 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5521 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5581 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5641 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5701 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5761 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5821 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5881 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5941 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6001 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6061 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6121 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6301 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6361 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6421 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6481 cgaggccctt tcgtcttcaa gaattagctt ttcaattcat catttttttt ttattctttt
     6541 ttttgatttc ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     6601 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     6661 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacatgcag gaaacgaaga
     6721 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     6781 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     6841 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     6901 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     6961 gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt
     7021 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     7081 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     7141 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     7201 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     7261 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     7321 attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga
     7381 accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt
     7441 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     7501 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     7561 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccaa ttctcatgtt
     7621 tgacagctta tcatcgatcg tccaactgca tggagatgag tcgtggcaag aataccaaga
     7681 gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca acatactact
     7741 cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag aagcaggtgg
     7801 gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc aagagagccc
     7861 cgagagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg gtgatgcgct
     7921 tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa atggtgtaaa
     7981 agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta ttactgagta
     8041 gtatttattt aagtattgtt tgtgcacttg cctgcaagcc ttttgaaaag caagcataaa
     8101 agatctaaac ataaaatctg taaaataaca agatgtaaag ataatgctaa atcatttggc
     8161 tttttgattg attgtacagg aaaatataca tcgcaggggg ttgactttta ccatttcacc
     8221 gcaatggaat caaacttgtt gaagagaatg ttcacaggcg catacgctac aatgacccga
     8281 ttcttgctag ccttttctcg gtcttgcaaa caaccgccgg cagcttagta tataaataca
     8341 catgtacata cctctctccg tatcctcgta atcattttct tgtatttatc gtcttttcgc
     8401 tgtaaaaact ttatcacact tatctcaaat acacttatta accgctttta ctattatctt
     8461 ctacgctgac agtaatatca aacagtgaca catattaaac acagtggttt ctttgcataa
     8521 acaccatcag cctcaagtcg tcaagtaaag atttcgtgtt catgcagata gataacaatc
     8581 tatatgttga taattagcgt tgcctcatca atgcgagatc cgtttaaccg gaccctagtg
     8641 cacttacccc acgttcggtc cactgtgtgc cgaacatgct ccttcactat tttaacatgt
     8701 ggaattaatt ctcatgtttg acagcttatc atcgaactct aagaggtgat acttatttac
     8761 tgtaaaactg tgacgataaa accggaagga agaataagaa aactcgaact gatctataat
     8821 gcctattttc tgtaaagagt ttaagctatg aaagcctcgg cattttggcc gctcctaggt
     8881 agtgcttttt ttccaaggac aaaacagttt ctttttcttg agcaggtttt atgtttcggt
     8941 aatcataaac aataaataaa ttatttcatt tatgtttaaa aataaaaaat aaaaaagtat
     9001 tttaaatttt taaaaaagtt gattataagc atgtgacctt ttgcaagcaa ttaaattttg
     9061 caatttgtga tttaggcaaa agttactatt tctggctcgt gtaatatatg tatgctaatg
     9121 tgaactttta caaagtcgat atggacttag tcaaaagaaa ttttcttaaa aatatatagc
     9181 actagccaat ttagcacttc tttatgagat atattataga ctttattaag ccagatttgt
     9241 gtattatatg tatttacccg gcgaatcatg gacatacatt ctgaaatagg taatattctc
     9301 tatggtgaga cagcatagat aacctaggat acaagttaaa agctagtact gttttgcagt
     9361 aatttttttc ttttttataa gaatgttacc acctaaataa gttataaagt caatagttaa
     9421 gtttgatatt tgattgtaaa ataccgtaat atatttgcat gatcaaaagg ctcaatgttg
     9481 actagccagc atgtcaacca ctatattgat caccgatatt aggacttcca caccaactag
     9541 taatatgaca ataaattcaa gatattcttc atgagaatgg cccagcgatt atgcggtgtg
     9601 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     9661 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     9721 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     9781 gtt
//
