LOCUS       sequence-460222-        6563 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses p53 mutant CASM203 in mamallian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6563)
  TITLE     A cancer-associated TP53 synonymous mutation induces synthesis of 
            the p53 isoform p53/47
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3-p53-G609T
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(16..219)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(45..65)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        220..599
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     771..790
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        865..969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(970..1038,1039..1830)
                     /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     complement(1188..1207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2001..2589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2490..2509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2743..2760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2877..2898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2913..2918,2919..2936,2937..2943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2951..2967
                     /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)."
     primer_bind     2956..2978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2975..2991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2975..2991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3040..3161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3051..3070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3105..3124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3335..4129)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3446..3465)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4056..4075
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4196..4525)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4210..4345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4264..4283)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4510..4530
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4539..4967)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4649..4670)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4861..4880
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5013..5237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5226..5243
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        5263..5281
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5263..5280
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(5315..6496)
                     /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
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(6515..6534)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6516..6534)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tctagttagc cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
       61 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
      121 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
      181 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
      241 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
      301 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
      361 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
      421 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
      481 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
      541 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
      601 acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt
      661 cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta
      721 ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga
      781 gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca
      841 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      901 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      961 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1021 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1081 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1141 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1201 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1261 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1321 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1381 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1441 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1501 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1561 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1621 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1681 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1741 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1801 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1861 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1921 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1981 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2041 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2101 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2161 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2221 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2281 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2341 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2401 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2461 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2521 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2581 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2641 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2701 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2761 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2821 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2881 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     2941 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     3001 gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg
     3061 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3121 ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc
     3181 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     3241 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc
     3301 gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     3361 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     3421 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     3481 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     3541 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     3601 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     3661 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     3721 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     3781 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     3841 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     3901 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     3961 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     4021 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     4081 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     4141 ctcatcctgt ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc
     4201 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg
     4261 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     4321 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     4381 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     4441 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     4501 actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta
     4561 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4621 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4681 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4741 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4801 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4861 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4921 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct
     4981 agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg
     5041 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     5101 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     5161 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     5221 gcaactagaa ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag
     5281 aatagggccc tctagatgca tgctcgagtt actatcagtc tgagtcaggc ccttctgtct
     5341 tgaacatgag ttttttatgg cgggaggtag actgaccctt tttggacttc aggtggctgg
     5401 agtgagccct gctcccccct ggctccttcc cagcctgggc atccttgagt tccaaggcct
     5461 cattcagctc tcggaacatc tcgaagcgct cacgcccacg gatctgaagg gtgaaatatt
     5521 ctccatccag tggtttcttc tttggctggg gagaggagct ggtgttgttg ggcagtgctc
     5581 gcttagtgct ccctgggggc agctcgtggt gaggctcccc tttcttgcgg agattctctt
     5641 cctctgtgcg ccggtctctc ccaggacagg cacaaacacg cacctcaaag ctgttccgtc
     5701 ccagtagatt accactggag tcttccagtg tgatgatggt gaggatgggc ctccggttca
     5761 tgccgcccat gcaggaactg ttacacatgt agttgtagtg gatggtggta cagtcagagc
     5821 caacctcagg cggctcatag ggcaccacca cactatgtcg aaaagtgttt ctgtcatcca
     5881 aatactcaac acgcaaattt ccttccactc ggataagatg ctgaggaggg gccagaccat
     5941 cgctatctga gcagcgctca tggtgggggc agcgcctcac aacctccgtc atgtgctgtg
     6001 actgcttgta gatggccatg gcgcggacgc gggtgccggg cgggggtgtg gaatcaaccc
     6061 acagctgcac agggcaggtc ttggccagtt ggcaaaacat cttgttgagg gcaggggagt
     6121 acgtgcaagt cacagacttg gctgtcccag aatgcaagaa gcccagacgg aaaccgtagc
     6181 tgccctggta ggttttctgg gaagggacag aagatgacag gggccaggag ggggctggtg
     6241 caggggccgc cggtgtagga gctgctggtg caggggccac ggggggagca gcctctggca
     6301 ttctgggagc ttcatctgga cctgggtctt cagtgaacca ttgttcaata tcgtccgggg
     6361 acagcatcaa atcatccatt gcttgggacg gcaaggggga cagaacgttg ttttcaggaa
     6421 gtagtttcca taggtctgaa aatgtttcct gactcagagg gggctcgacg ctaggatctg
     6481 actgcggctc ctccatggca gtaagcttgg gtctccctat agtgagtcgt attaatttcg
     6541 ataagccagt aagcagtggg ttc
//
