LOCUS       sequence-176917-        8477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression of mouse PKC Eta with yeast PKC1 promoter and ADH1 
            terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8477)
  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 8477)
  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 Eta (JCE615)
FEATURES             Location/Qualifiers
     source          1..8477
                     /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      2721..2908
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2776..2800)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2968..2984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2968..2984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2981..3003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2992..3008
                     /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(3016..3022,3023..3040,3041..3046))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3061..3082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3199..3216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3370..3958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3450..3469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4129..4920,4921..4989))
                     /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     4752..4771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4990..5094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5162..5180
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5205..5420
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5421..6224
                     /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    6559..7721
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      6559..7396
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     join(8463..8477,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8477,1..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 gtcggtaccc gggatgtcgt ccggcacgat gaagttcaat
      661 ggctatctga gggtccgcat cggagaggct gtagggctgc agcccacccg ctggtccctg
      721 cggcactcgc tcttcaaaaa gggccaccag ctgctggacc cctacctgac ggtgagcgta
      781 gaccaggtac gcgtgggcca gaccagcaca aagcagaaga ccaacaaacc cacctacaac
      841 gaggagttct gcgccaatgt caccgacggc ggccacctgg agctagccgt cttccacgag
      901 acgcccctgg gttatgacca ctttgtggcc aactgcacgc tgcagttcca ggagctgttg
      961 cgcacggctg gtacctcgga caccttcgag ggctgggtgg atctggagcc tgaggggaaa
     1021 gtgtttgtgg taataaccct aacagggagt ttcactgaag ccactctcca gagagaccgc
     1081 atcttcaagc attttaccag gaagcgccaa agggctatgc gaagacgagt ccatcaagtg
     1141 aacggacata agttcatggc cacgtacctg aggcagccca cctactgctc tcattgccga
     1201 gagttcatct ggggagtatt tgggaaacag ggttatcaat gccaagtgtg cacctgcgtc
     1261 gtccataaac gctgccatca tctaattgtt acagcctgca cttgccaaaa caatattaac
     1321 aaagtggatg ccaagattgc agaacagcgg tttggcatca acatcccaca caagttcaac
     1381 gttcacaatt acaaggtgcc cacgttctgt gaccactgtg gctccctgct ctgggggata
     1441 atgcgacaag gacttcagtg taaaatatgt aagatgaatg tacatattcg gtgtcaggcg
     1501 aacgtggccc caaactgcgg ggtgaatgcc gtggagcttg ccaagaccct ggcagggatg
     1561 ggtctccaac ccggaaatat ttctccaacc tcgaaactca tttccaggtc gacactaaga
     1621 cggcagggaa aggagggctc caaagaagga aatgggatcg gtgttaactc ttccagcaga
     1681 ttcggcatcg acaactttga gttcatccgg gtgttgggga aggggagctt cgggaaggtg
     1741 atgcttgcca ggataaagga gacaggagaa ctgtacgccg tgaaggtgct gaagaaggac
     1801 gtgattctgc aggatgatga tgtagagtgc accatgactg agaagaggat cctgtccttg
     1861 gcccgcaacc accccttcct cacccagctc ttctgctgct ttcagactcc agaccgtctg
     1921 ttctttgtca tggagtttgt gaacggaggc gacctgatgt tccacatcca aaagtcccgt
     1981 cgtttcgatg aagcccgtgc tcgtttctac gccgcggaga tcatttctgc actcatgttc
     2041 ctacatgaga aaggtatcat ctatagagac ttgaaactgg acaatgtgct attggaccac
     2101 gaaggtcact gtaaactggc cgactttgga atgtgcaagg aggggatttg taatggggtc
     2161 accacagcca ccttctgcgg tacacccgac tacattgccc cggagatcct tcaggagatg
     2221 ctgtatggac ctgcagtaga ctggtgggcc atgggtgtgt tgctttatga gatgctgtgt
     2281 ggacatgcgc cctttgaggc tgaaaatgaa gatgaccttt ttgaggccat actgaatgat
     2341 gaagtcgtct accccacctg gctccacgaa gatgccacag ggatcctcaa gtctttcatg
     2401 accaagaacc ccaccatgcg cttgggcagc ctgactcagg gaggagagca tgagatcctg
     2461 agacaccctt tctttaagga aatcgactgg gcccagttga accatcgcca gttagagccg
     2521 cctttccgac ctagaatcaa atcccgagaa gatgtcagca attttgaccc agactttata
     2581 aaagaagagc ccgtcttaac tccgattgat gagggacatc ttcctatgat taaccaggat
     2641 gagtttagaa acttttccta tgtgtcaccg gaattgcaac tgtagtctag agtcgactag
     2701 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     2761 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     2821 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
     2881 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatatctg
     2941 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3001 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3061 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3121 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3181 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3241 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3301 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3361 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3421 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3481 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3541 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3601 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3661 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3721 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3781 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3841 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3901 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3961 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4021 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4081 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5101 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5161 aaataggcgt atcacgaggc cctttcgtct tcaagaatta gcttttcaat tcatcatttt
     5221 ttttttattc ttttttttga tttcggtttc cttgaaattt ttttgattcg gtaatctccg
     5281 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     5341 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacat
     5401 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     5461 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     5521 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     5581 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     5641 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa gacagaaaat
     5701 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     5761 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     5821 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     5881 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     5941 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     6001 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gacgcattgg
     6061 gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt attattgttg
     6121 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     6181 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     6241 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     6301 ccaattctca tgtttgacag cttatcatcg atcgtccaac tgcatggaga tgagtcgtgg
     6361 caagaatacc aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac
     6421 tgcaacatac tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat
     6481 tcagaagcag gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga
     6541 aggcaagaga gccccgagag cttacatttt atgttagctg gtggactgac gccagaaaat
     6601 gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag
     6661 acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag
     6721 gttattactg agtagtattt atttaagtat tgtttgtgca cttgcctgca agccttttga
     6781 aaagcaagca taaaagatct aaacataaaa tctgtaaaat aacaagatgt aaagataatg
     6841 ctaaatcatt tggctttttg attgattgta caggaaaata tacatcgcag ggggttgact
     6901 tttaccattt caccgcaatg gaatcaaact tgttgaagag aatgttcaca ggcgcatacg
     6961 ctacaatgac ccgattcttg ctagcctttt ctcggtcttg caaacaaccg ccggcagctt
     7021 agtatataaa tacacatgta catacctctc tccgtatcct cgtaatcatt ttcttgtatt
     7081 tatcgtcttt tcgctgtaaa aactttatca cacttatctc aaatacactt attaaccgct
     7141 tttactatta tcttctacgc tgacagtaat atcaaacagt gacacatatt aaacacagtg
     7201 gtttctttgc ataaacacca tcagcctcaa gtcgtcaagt aaagatttcg tgttcatgca
     7261 gatagataac aatctatatg ttgataatta gcgttgcctc atcaatgcga gatccgttta
     7321 accggaccct agtgcactta ccccacgttc ggtccactgt gtgccgaaca tgctccttca
     7381 ctattttaac atgtggaatt aattctcatg tttgacagct tatcatcgaa ctctaagagg
     7441 tgatacttat ttactgtaaa actgtgacga taaaaccgga aggaagaata agaaaactcg
     7501 aactgatcta taatgcctat tttctgtaaa gagtttaagc tatgaaagcc tcggcatttt
     7561 ggccgctcct aggtagtgct ttttttccaa ggacaaaaca gtttcttttt cttgagcagg
     7621 ttttatgttt cggtaatcat aaacaataaa taaattattt catttatgtt taaaaataaa
     7681 aaataaaaaa gtattttaaa tttttaaaaa agttgattat aagcatgtga ccttttgcaa
     7741 gcaattaaat tttgcaattt gtgatttagg caaaagttac tatttctggc tcgtgtaata
     7801 tatgtatgct aatgtgaact tttacaaagt cgatatggac ttagtcaaaa gaaattttct
     7861 taaaaatata tagcactagc caatttagca cttctttatg agatatatta tagactttat
     7921 taagccagat ttgtgtatta tatgtattta cccggcgaat catggacata cattctgaaa
     7981 taggtaatat tctctatggt gagacagcat agataaccta ggatacaagt taaaagctag
     8041 tactgttttg cagtaatttt tttctttttt ataagaatgt taccacctaa ataagttata
     8101 aagtcaatag ttaagtttga tatttgattg taaaataccg taatatattt gcatgatcaa
     8161 aaggctcaat gttgactagc cagcatgtca accactatat tgatcaccga tattaggact
     8221 tccacaccaa ctagtaatat gacaataaat tcaagatatt cttcatgaga atggcccagc
     8281 gattatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     8341 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     8401 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     8461 ttcccagtca cgacgtt
//
