LOCUS       sequence-136118-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains the CORE cassette with hyg and human p53 gene markers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6319)
  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 6319)
  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-Hp53
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            142..1717
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        142..485
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             486..1514
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             486..1514
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1520..1717
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        1916..2580
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2132..2249
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2544..2567
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     RBS             2612..2634
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2644..3825
                     /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(3956..3975)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3957..3975)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4062..4079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4233..4821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4313..4332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4992..5783,5784..5852))
                     /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     5615..5634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5853..5957)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6025..6043
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6081..6103)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6203..6222
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6303..6319,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6303..6319,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatccccggg ttaattaagg cgcgccagat ctgtttagct tgcctcgtcc
      121 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
      181 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
      241 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
      301 tcctcgctgc ggacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
      361 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
      421 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
      481 caaccatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
      541 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
      601 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
      661 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
      721 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
      781 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
      841 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
      901 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
      961 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     1021 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     1081 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     1141 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     1201 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     1261 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     1321 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     1381 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     1441 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     1501 gggcaaagga ataatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     1561 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     1621 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     1681 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     1741 ccatccagtg tcgaaaacga gctcgaattc gacaggttat cagcaacaac acagtcatat
     1801 ccattctcaa ttagctctac cacagtgtgt gaaccaatgt atccagcacc acctgtaacc
     1861 aaaacaattt tagaagtact ttcactttgt aactgagctg tcatttatat tgaattttca
     1921 aaaattctta cttttttttt ggatggacgc aaagaagttt aataatcata ttacatggca
     1981 ttaccaccat atacatatcc atatacatat ccatatctaa tcttacttat atgttgtgga
     2041 aatgtaaaga gccccattat cttagcctaa aaaaaccttc tctttggaac tttcagtaat
     2101 acgcttaact gctcattgct atattgaagt acggattaga agccgccgag cgggtgacag
     2161 ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac
     2221 gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta
     2281 tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa atgaacgaat
     2341 caaattaaca accataggat gataatgcga ttagtttttt agccttattt ctggggtaat
     2401 taatcagcga agcgatgatt tttgatctat taacagatat ataaatgcaa aaactgcata
     2461 accactttaa ctaatacttt caacattttc ggtttgtatt acttcttatt caaatgtaat
     2521 aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaaac
     2581 cccggatccg gccccctcga ctagaaataa ttttgtttaa ctttaagaag gagatatagc
     2641 atgatggagg agccgcagtc agatcctagc gtcgagcccc ctctgagtca ggaaacattt
     2701 tcagacctat ggaaactact tcctgaaaac aacgttctgt cccccttgcc gtcccaagca
     2761 atggatgatt tgatgctgtc cccggacgat attgaacaat ggttcactga agacccaggt
     2821 ccagatgaag ctcccagaat gccagaggct gctccccccg tggcccctgc accagcagct
     2881 cctacaccgg cggcccctgc accagccccc tcctggcccc tgtcatcttc tgtcccttcc
     2941 cagaaaacct accagggcag ctacggtttc cgtctgggct tcttgcattc tgggacagcc
     3001 aagtctgcga cttgcacgta ctcccctgcc ctcaacaaga tgttttgcca actggccaag
     3061 acctgccctg tgcagctgtg ggttgattcc acacccccgc ccggcacccg cgtccgcgcc
     3121 atggccatct acaagcagtc acagcacatg acggaggttg tgaggcgctg cccccaccat
     3181 gagcgctgct cagatagcga tggtctggcc cctcctcagc atcttatccg agtggaagga
     3241 aatttgcgtg tggagtattt ggatgacaga aacacttttc gacatagtgt ggtggtgccc
     3301 tatgagccgc ctgaggttgg ctctgactgt accaccatcc actacaacta catgtgtaac
     3361 agttcctgca tgggcggcat gaaccggagg cccatcctca ccatcatcac actggaagac
     3421 tccagtggta atctactggg acggaacagc tttgaggtgc gtgtttgtgc ctgtcctggg
     3481 agagaccggc gcacagagga agagaatctc cgcaagaaag gggagcctca ccacgagctg
     3541 cccccaggga gcactaagcg agcactgccc aacaacacca gctcctctcc ccagccaaag
     3601 aagaaaccac tggatggaga atatttcacc cttcagatcc gtgggcgtga gcgcttcgag
     3661 atgttccgag agctgaatga ggccttggaa ctcaaggatg cccaggctgg gaaggagcca
     3721 ggggggagca gggctcactc cagccacctg aagtccaaaa agggtcagtc tacctcccgc
     3781 cataaaaaac tcatgttcaa gacagaaggg cctgactcag actgacattc tccacttctt
     3841 gttccccact gacagcctcc cacccccatc tctccctccc ctgccatttt gggtttccct
     3901 taattaatta agggggatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta
     3961 tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc
     4021 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     4441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5101 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5221 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5281 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5341 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5401 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5461 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5521 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5581 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5641 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5701 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5761 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5821 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5881 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5941 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6001 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6061 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     6121 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6181 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     6241 gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata
     6301 cgatttaggt gacactata
//
