LOCUS       sequence-361960-        8203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression of mouse PKC Iota with yeast PKC1 promoter and ADH1
            terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8203)
  TITLE     Yeast expression plasmids of mouse PKC isoforms with yeast PKC1 
            promoter and ADH1 terminator
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8203)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: isoform PKC Iota (JCE597)
FEATURES             Location/Qualifiers
     source          1..8203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(108..212)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     280..298
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        323..538
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             539..1342
                     /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    1677..2839
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      1677..2514
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     3581..3603
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3595..3612
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3596..3612
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6042..6229
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6097..6121)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(6289..6305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6289..6305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6302..6324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6313..6329
                     /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(6337..6343,6344..6361,6362..6367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6382..6403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6520..6537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6691..7279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6771..6790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7450..8203,1..38,39..107))
                     /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     8073..8092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
       61 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
      181 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      241 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      301 tttcgtcttc aagaattagc ttttcaattc atcatttttt ttttattctt ttttttgatt
      361 tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
      421 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
      481 cccagtattc ttaacccaac tgcacagaac aaaaacatgc aggaaacgaa gataaatcat
      541 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
      601 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
      661 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
      721 tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc
      781 cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt
      841 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
      901 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
      961 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
     1021 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     1081 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     1141 acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga
     1201 tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg
     1261 aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag
     1321 aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc
     1381 acaaattaga gcttcaattt aattatatca gttattaccc aattctcatg tttgacagct
     1441 tatcatcgat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
     1501 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
     1561 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
     1621 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgagagct
     1681 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     1741 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     1801 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     1861 ttaagtattg tttgtgcact tgcctgcaag ccttttgaaa agcaagcata aaagatctaa
     1921 acataaaatc tgtaaaataa caagatgtaa agataatgct aaatcatttg gctttttgat
     1981 tgattgtaca ggaaaatata catcgcaggg ggttgacttt taccatttca ccgcaatgga
     2041 atcaaacttg ttgaagagaa tgttcacagg cgcatacgct acaatgaccc gattcttgct
     2101 agccttttct cggtcttgca aacaaccgcc ggcagcttag tatataaata cacatgtaca
     2161 tacctctctc cgtatcctcg taatcatttt cttgtattta tcgtcttttc gctgtaaaaa
     2221 ctttatcaca cttatctcaa atacacttat taaccgcttt tactattatc ttctacgctg
     2281 acagtaatat caaacagtga cacatattaa acacagtggt ttctttgcat aaacaccatc
     2341 agcctcaagt cgtcaagtaa agatttcgtg ttcatgcaga tagataacaa tctatatgtt
     2401 gataattagc gttgcctcat caatgcgaga tccgtttaac cggaccctag tgcacttacc
     2461 ccacgttcgg tccactgtgt gccgaacatg ctccttcact attttaacat gtggaattaa
     2521 ttctcatgtt tgacagctta tcatcgaact ctaagaggtg atacttattt actgtaaaac
     2581 tgtgacgata aaaccggaag gaagaataag aaaactcgaa ctgatctata atgcctattt
     2641 tctgtaaaga gtttaagcta tgaaagcctc ggcattttgg ccgctcctag gtagtgcttt
     2701 ttttccaagg acaaaacagt ttctttttct tgagcaggtt ttatgtttcg gtaatcataa
     2761 acaataaata aattatttca tttatgttta aaaataaaaa ataaaaaagt attttaaatt
     2821 tttaaaaaag ttgattataa gcatgtgacc ttttgcaagc aattaaattt tgcaatttgt
     2881 gatttaggca aaagttacta tttctggctc gtgtaatata tgtatgctaa tgtgaacttt
     2941 tacaaagtcg atatggactt agtcaaaaga aattttctta aaaatatata gcactagcca
     3001 atttagcact tctttatgag atatattata gactttatta agccagattt gtgtattata
     3061 tgtatttacc cggcgaatca tggacataca ttctgaaata ggtaatattc tctatggtga
     3121 gacagcatag ataacctagg atacaagtta aaagctagta ctgttttgca gtaatttttt
     3181 tcttttttat aagaatgtta ccacctaaat aagttataaa gtcaatagtt aagtttgata
     3241 tttgattgta aaataccgta atatatttgc atgatcaaaa ggctcaatgt tgactagcca
     3301 gcatgtcaac cactatattg atcaccgata ttaggacttc cacaccaact agtaatatga
     3361 caataaattc aagatattct tcatgagaat ggcccagcga ttatgcggtg tgaaataccg
     3421 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     3481 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     3541 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     3601 acgacggcca gtgaattcga gtggttcagc ttctgtttga gttgcaacca tggtatgcat
     3661 gccggtcacg ctgaagaatg gtttgacaga cataatgttt gtcccactcc aggttgcacc
     3721 tgccagtgta ataagtagta ttgtacaata aaccaaatct tgcaacttct ctttattctc
     3781 acaatatata tacataacca aagaaaaaac gtaattagta taacattgta tttatgaggc
     3841 gttgctatct tcattaccca gcccgcctag ttttaaaggc gaaaaaaacc gacgaaaaga
     3901 gttaacaagc gtggataata aagttgatct actctatcga aagtaatacg acaggttcag
     3961 gagcagtaaa agagctacga tttcactgct attcaatttg gctgagtaga gactgattat
     4021 taaccattgt aaaagtctct gccaccactc gtccacactt acgttagagg tattcagcgt
     4081 cgatcccctt tcagtataga tttttaaaac tatattagat aatcaatagg aataaaacct
     4141 agaacagtag ctgcggagag aagctctagc gaggaaaagt aagtatagta tcacacatat
     4201 agggagcagt ttacagtcgg taccatgccg acccagaggg acagcagcac catgtcccac
     4261 acggtcgcgt gcggcggcgg cggggaccat tcccaccagg tccgggtgaa agcctactac
     4321 cgcggggata ttatgataac acactttgag ccttccatct cctttgaggg actttgcagt
     4381 gaggttcgag atatgtgttc ttttgacaat gagcagccgt tcaccatgaa atggatagat
     4441 gaggaaggag acccatgtac agtgtcttct cagttggagt tagaagaggc tttcaggctg
     4501 tacgagctga acaaggattc tgaactcttg attcatgtat ttccatgtgt accagagcgt
     4561 cctggaatgc cttgcccagg ggaagacaag tccatttacc ggagaggggc acgccggtgg
     4621 agaaagctgt attgtgcaaa tggccacact tttcaagcca aacgtttcaa taggcgcgcc
     4681 cactgtgcca tctgcacaga cagaatctgg ggcctcggac gacaaggata caagtgcatc
     4741 aactgcaaac tgctggttca taagaagtgc cacaagctgg tcacaattga gtgtgggcgg
     4801 cactctttgc caccggaacc catgatgcca atggaccaga ccatgcatcc agaccacaca
     4861 cagacagtaa ttccatataa tccttcaagt catgagagtt tggaccaagt tggtgaagaa
     4921 aaggaggcaa tgaacaccag ggagagtggt aaagcgtcgt ccagtctagg tctgcaggat
     4981 ttcgatttgc ttcgagttat agggagggga agttacgcca aagtactgtt ggttcggtta
     5041 aagaaaacag atcgcattta tgcaatgaaa gttgtgaaga aagagctcgt caatgacgat
     5101 gaggatatcg attgggtgca gacagagaag catgtgtttg agcaggcgtc caatcaccct
     5161 tttcttgtcg ggctgcattc ttgcttccag acagaaagca ggttgttttt tgtcatagaa
     5221 tatgtaaatg gaggggacct catgtttcat atgcagcgac agagaaaact tcctgaagag
     5281 catgccaggt tttactctgc agaaatcagt ctagcactga attatcttca tgagcgaggg
     5341 ataatttata gagatttgaa gttggacaat gtactgctag actctgaagg acacattaaa
     5401 ctcactgact acggcatgtg taaggaagga ttgcggcctg gagacacaac cagcactttc
     5461 tgcggcactc ccaattacat tgctccagag atcttaagag gagaagatta tggcttcagc
     5521 gttgactggt gggctcttgg agtgcttatg tttgagatga tggcgggaag gtctccgttt
     5581 gatatcgttg ggagctctga caatcctgac caaaacacag aggattatct attccaagtc
     5641 attttggaaa agcagatccg catcccgcgt tctctgtctg taaaagcagc aagtgtactg
     5701 aagagttttc tcaacaagga cccaaaggaa cgattgggtt gtcaccctca aactggattt
     5761 gctgacattc aaggacatcc attcttcaga aatgtggact gggacatgat ggagcaaaag
     5821 caggtggttc caccctttaa accaaacatt tctggagaat ttggtttgga taatttcgat
     5881 tctcagttta ctaatgaacc agtccagctc actccagatg atgatgacat tgtgaggaag
     5941 attgatcagt ctgaatttga aggttttgag tatatcaacc ccctcttgat gtctgcagaa
     6001 gagtgtgtct gagtcgacta gggcgcgcca cttctaaata agcgaatttc ttatgattta
     6061 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     6121 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     6181 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta
     6241 gggataacag ggtaatatct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     6301 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6361 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6421 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7561 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7741 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7921 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tga
//
