LOCUS       sequence-177664-        8176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression of mouse PKC lambda with yeast PKC1 promoter and 
            ADH1 terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8176)
  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 8176)
  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 lambda (JCE590)
FEATURES             Location/Qualifiers
     source          1..8176
                     /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      2420..2607
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2475..2499)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2667..2683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2667..2683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2680..2702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2691..2707
                     /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(2715..2721,2722..2739,2740..2745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2760..2781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2898..2915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3069..3657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3149..3168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3828..4619,4620..4688))
                     /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     4451..4470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4689..4793)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4861..4879
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4904..5119
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5120..5923
                     /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    6258..7420
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      6258..7095
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     join(8162..8176,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8176,1..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 tgtcccacac ggtcgcgtgc ggcggcggcg
      661 gggaccattc ccaccaggtc cgggtgaaag cctactaccg cggggatatt atgataacac
      721 actttgagcc ttccatctcc tttgagggac tttgcagtga ggttcgagat atgtgttctt
      781 ttgacaatga gcagccgttc accatgaaat ggatagatga ggaaggagac ccatgtacag
      841 tgtcttctca gttggagtta gaagaggctt tcaggctgta cgagctgaac aaggattctg
      901 aactcttgat tcatgtattt ccatgtgtac cagagcgtcc tggaatgcct tgcccagggg
      961 aagacaagtc catttaccgg agaggggcac gccggtggag aaagctgtat tgtgcaaatg
     1021 gccacacttt tcaagccaaa cgtttcaata ggcgcgccca ctgtgccatc tgcacagaca
     1081 gaatctgggg cctcggacga caaggataca agtgcatcaa ctgcaaactg ctggttcata
     1141 agaagtgcca caagctggtc acaattgagt gtgggcggca ctctttgcca ccggaaccca
     1201 tgatgccaat ggaccagacc atgcatccag accacacaca gacagtaatt ccatataatc
     1261 cttcaagtca tgagagtttg gaccaagttg gtgaagaaaa ggaggcaatg aacaccaggg
     1321 agagtggtaa agcgtcgtcc agtctaggtc tgcaggattt cgatttgctt cgagttatag
     1381 ggaggggaag ttacgccaaa gtactgttgg ttcggttaaa gaaaacagat cgcatttatg
     1441 caatgaaagt tgtgaagaaa gagctcgtca atgacgatga ggatatcgat tgggtgcaga
     1501 cagagaagca tgtgtttgag caggcgtcca atcacccttt tcttgtcggg ctgcattctt
     1561 gcttccagac agaaagcagg ttgttttttg tcatagaata tgtaaatgga ggggacctca
     1621 tgtttcatat gcagcgacag agaaaacttc ctgaagagca tgccaggttt tactctgcag
     1681 aaatcagtct agcactgaat tatcttcatg agcgagggat aatttataga gatttgaagt
     1741 tggacaatgt actgctagac tctgaaggac acattaaact cactgactac ggcatgtgta
     1801 aggaaggatt gcggcctgga gacacaacca gcactttctg cggcactccc aattacattg
     1861 ctccagagat cttaagagga gaagattatg gcttcagcgt tgactggtgg gctcttggag
     1921 tgcttatgtt tgagatgatg gcgggaaggt ctccgtttga tatcgttggg agctctgaca
     1981 atcctgacca aaacacagag gattatctat tccaagtcat tttggaaaag cagatccgca
     2041 tcccgcgttc tctgtctgta aaagcagcaa gtgtactgaa gagttttctc aacaaggacc
     2101 caaaggaacg attgggttgt caccctcaaa ctggatttgc tgacattcaa ggacatccat
     2161 tcttcagaaa tgtggactgg gacatgatgg agcaaaagca ggtggttcca ccctttaaac
     2221 caaacatttc tggagaattt ggtttggata atttcgattc tcagtttact aatgaaccag
     2281 tccagctcac tccagatgat gatgacattg tgaggaagat tgatcagtct gaatttgaag
     2341 gttttgagta tatcaacccc ctcttgatgt ctgcagaaga gtgtgtctga gtcgactagg
     2401 gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt
     2461 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt
     2521 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt
     2581 cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg taatatctgc
     2641 aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2701 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2761 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2821 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2881 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2941 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3001 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3061 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3121 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3181 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3241 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3301 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3361 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3421 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3481 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3541 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3601 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3661 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3721 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3841 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3901 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3961 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4021 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4081 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4141 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4201 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4261 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4321 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4381 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4441 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4501 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4561 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4621 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4681 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4741 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4801 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     4861 aataggcgta tcacgaggcc ctttcgtctt caagaattag cttttcaatt catcattttt
     4921 tttttattct tttttttgat ttcggtttcc ttgaaatttt tttgattcgg taatctccga
     4981 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
     5041 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacatg
     5101 caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc
     5161 tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc
     5221 ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa
     5281 aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt
     5341 taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag acagaaaatt
     5401 tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga
     5461 atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa
     5521 gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc
     5581 atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag
     5641 cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg
     5701 ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag acgcattggg
     5761 tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta ttattgttgg
     5821 aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc
     5881 aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt
     5941 aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc agttattacc
     6001 caattctcat gtttgacagc ttatcatcga tcgtccaact gcatggagat gagtcgtggc
     6061 aagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt ccaaaagact
     6121 gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc ttgtttgatt
     6181 cagaagcagg tgggacaggt gaacttttgg attggaactc gatttctgac tgggttggaa
     6241 ggcaagagag ccccgagagc ttacatttta tgttagctgg tggactgacg ccagaaaatg
     6301 ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga
     6361 caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct aagaaatagg
     6421 ttattactga gtagtattta tttaagtatt gtttgtgcac ttgcctgcaa gccttttgaa
     6481 aagcaagcat aaaagatcta aacataaaat ctgtaaaata acaagatgta aagataatgc
     6541 taaatcattt ggctttttga ttgattgtac aggaaaatat acatcgcagg gggttgactt
     6601 ttaccatttc accgcaatgg aatcaaactt gttgaagaga atgttcacag gcgcatacgc
     6661 tacaatgacc cgattcttgc tagccttttc tcggtcttgc aaacaaccgc cggcagctta
     6721 gtatataaat acacatgtac atacctctct ccgtatcctc gtaatcattt tcttgtattt
     6781 atcgtctttt cgctgtaaaa actttatcac acttatctca aatacactta ttaaccgctt
     6841 ttactattat cttctacgct gacagtaata tcaaacagtg acacatatta aacacagtgg
     6901 tttctttgca taaacaccat cagcctcaag tcgtcaagta aagatttcgt gttcatgcag
     6961 atagataaca atctatatgt tgataattag cgttgcctca tcaatgcgag atccgtttaa
     7021 ccggacccta gtgcacttac cccacgttcg gtccactgtg tgccgaacat gctccttcac
     7081 tattttaaca tgtggaatta attctcatgt ttgacagctt atcatcgaac tctaagaggt
     7141 gatacttatt tactgtaaaa ctgtgacgat aaaaccggaa ggaagaataa gaaaactcga
     7201 actgatctat aatgcctatt ttctgtaaag agtttaagct atgaaagcct cggcattttg
     7261 gccgctccta ggtagtgctt tttttccaag gacaaaacag tttctttttc ttgagcaggt
     7321 tttatgtttc ggtaatcata aacaataaat aaattatttc atttatgttt aaaaataaaa
     7381 aataaaaaag tattttaaat ttttaaaaaa gttgattata agcatgtgac cttttgcaag
     7441 caattaaatt ttgcaatttg tgatttaggc aaaagttact atttctggct cgtgtaatat
     7501 atgtatgcta atgtgaactt ttacaaagtc gatatggact tagtcaaaag aaattttctt
     7561 aaaaatatat agcactagcc aatttagcac ttctttatga gatatattat agactttatt
     7621 aagccagatt tgtgtattat atgtatttac ccggcgaatc atggacatac attctgaaat
     7681 aggtaatatt ctctatggtg agacagcata gataacctag gatacaagtt aaaagctagt
     7741 actgttttgc agtaattttt ttctttttta taagaatgtt accacctaaa taagttataa
     7801 agtcaatagt taagtttgat atttgattgt aaaataccgt aatatatttg catgatcaaa
     7861 aggctcaatg ttgactagcc agcatgtcaa ccactatatt gatcaccgat attaggactt
     7921 ccacaccaac tagtaatatg acaataaatt caagatattc ttcatgagaa tggcccagcg
     7981 attatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     8041 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8101 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     8161 tcccagtcac gacgtt
//
