LOCUS       sequence-136109-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains the CORE cassette with KanMX4 and human p53 gene markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  AUTHORS   Storici F, Resnick MA
  TITLE     Delitto perfetto targeted mutagenesis in yeast with 
            oligonucleotides.
  JOURNAL   Genet Eng (N Y). 2003;25:189-207.
  PUBMED    15260239
REFERENCE   2  (bases 1 to 6073)
  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: "Genet Eng 
            (N Y)."; date: "2003"; volume: "25"; pages: "189-207"
COMMENT     Sequence Label: pCORE-Kp53
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            115..1471
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        115..458
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             459..1268
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(526..545)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1274..1471
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        1670..2334
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    1886..2003
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2298..2321
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     RBS             2366..2388
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2398..3579
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSATCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(3710..3729)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3711..3729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3816..3833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4067..4086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4746..5537,5538..5606))
                     /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     5369..5388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5607..5711)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5779..5797
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5835..5857)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5957..5976
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6057..6073,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6057..6073,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
      121 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      181 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      241 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
      301 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
      361 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
      421 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg
      481 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
      541 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
      601 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
      661 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
      721 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
      781 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
      841 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
      901 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
      961 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     1021 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     1081 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     1141 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     1201 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     1261 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     1321 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     1381 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     1441 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     1501 agtgtcgaaa acgagctcga attcgacagg ttatcagcaa caacacagtc atatccattc
     1561 tcaattagct ctaccacagt gtgtgaacca atgtatccag caccacctgt aaccaaaaca
     1621 attttagaag tactttcact ttgtaactga gctgtcattt atattgaatt ttcaaaaatt
     1681 cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat ggcattacca
     1741 ccatatacat atccatatac atatccatat ctaatcttac ttatatgttg tggaaatgta
     1801 aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag taatacgctt
     1861 aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggtg acagccctcc
     1921 gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat
     1981 gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta
     2041 tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac gaatcaaatt
     2101 aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca
     2161 gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg cataaccact
     2221 ttaactaata ctttcaacat tttcggtttg tattacttct tattcaaatg taataaaagt
     2281 atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa aaaccccgga
     2341 tccggccccc tcgactagaa ataattttgt ttaactttaa gaaggagata tagcatgatg
     2401 gaggagccgc agtcagatcc tagcgtcgag ccccctctga gtcaggaaac attttcagac
     2461 ctatggaaac tacttcctga aaacaacgtt ctgtccccct tgccgtccca agcaatggat
     2521 gatttgatgc tgtccccgga cgatattgaa caatggttca ctgaagaccc aggtccagat
     2581 gaagctccca gaatgccaga ggctgctccc cccgtggccc ctgcaccagc agctcctaca
     2641 ccggcggccc ctgcaccagc cccctcctgg cccctgtcat cttctgtccc ttcccagaaa
     2701 acctaccagg gcagctacgg tttccgtctg ggcttcttgc attctgggac agccaagtct
     2761 gcgacttgca cgtactcccc tgccctcaac aagatgtttt gccaactggc caagacctgc
     2821 cctgtgcagc tgtgggttga ttccacaccc ccgcccggca cccgcgtccg cgccatggcc
     2881 atctacaagc agtcacagca catgacggag gttgtgaggc gctgccccca ccatgagcgc
     2941 tgctcagata gcgatggtct ggcccctcct cagcatctta tccgagtgga aggaaatttg
     3001 cgtgtggagt atttggatga cagaaacact tttcgacata gtgtggtggt gccctatgag
     3061 ccgcctgagg ttggctctga ctgtaccacc atccactaca actacatgtg taacagttcc
     3121 tgcatgggcg gcatgaaccg gaggcccatc ctcaccatca tcacactgga agactccagt
     3181 ggtaatctac tgggacggaa cagctttgag gtgcgtgttt gtgcctgtcc tgggagagac
     3241 cggcgcacag aggaagagaa tctccgcaag aaaggggagc ctcaccacga gctgccccca
     3301 gggagcacta agcgagcact gcccaacaac accagctcct ctccccagcc aaagaagaaa
     3361 ccactggatg gagaatattt cacccttcag atccgtgggc gtgagcgctt cgagatgttc
     3421 cgagagctga atgaggcctt ggaactcaag gatgcccagg ctgggaagga gccagggggg
     3481 agcagggctc actccagcca cctgaagtcc aaaaagggtc agtctacctc ccgccataaa
     3541 aaactcatgt tcaagacaga agggcctgac tcagactgac attctccact tcttgttccc
     3601 cactgacagc ctcccacccc catctctccc tcccctgcca ttttgggttt cccttaatta
     3661 attaaggggg atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga
     3721 gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg
     3781 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     4201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4681 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4741 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4801 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4861 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4921 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4981 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5041 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5101 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5161 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5221 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5281 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5341 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5401 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5461 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5521 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5581 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5641 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5701 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5761 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5821 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5881 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5941 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata
     6001 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt
     6061 aggtgacact ata
//
