LOCUS       sequence-460223-        6563 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses p53 mutant G609A 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  JOURNAL   Unpublished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            "Unpublished"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-G609A
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     246..265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(484..588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(663..682)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        854..1233
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1234..1437
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1388..1408
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1482..1501
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1482..1500
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1520..2701
                     /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"
     promoter        complement(2735..2753)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2736..2753)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2773..2790)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2779..3003
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3049..3477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3136..3155)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3346..3367
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3486..3506)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3491..3820
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3671..3806
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3733..3752
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3887..4681
                     /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(3941..3960)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4551..4570
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4855..4976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4892..4911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4946..4965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5025..5041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5025..5041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5038..5060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5049..5065
                     /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(5073..5079,5080..5097,5098..5103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5118..5139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5256..5273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5427..6015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5507..5526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6186..6563,1..414,415..483))
                     /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"
ORIGIN
        1 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
       61 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      601 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
      661 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
      721 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
      781 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      841 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      901 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      961 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1021 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1081 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1141 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1201 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     1261 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     1321 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     1381 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     1441 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
     1501 gagacccaag cttactgcca tggaggagcc gcagtcagat cctagcgtcg agccccctct
     1561 gagtcaggaa acattttcag acctatggaa actacttcct gaaaacaacg ttctgtcccc
     1621 cttgccgtcc caagcaatgg atgatttgat gctgtccccg gacgatattg aacaatggtt
     1681 cactgaagac ccaggtccag atgaagctcc cagaatgcca gaggctgctc cccccgtggc
     1741 ccctgcacca gcagctccta caccggcggc ccctgcacca gccccctcct ggcccctgtc
     1801 atcttctgtc ccttcccaga aaacctacca gggcagctac ggtttccgtc tgggcttctt
     1861 gcattctggg acagccaagt ctgtgacttg cacgtactcc cctgccctca acaagatgtt
     1921 ttgccaactg gccaagacct gccctgtgca gctgtgggtt gattccacac ccccgcccgg
     1981 cacccgcgtc cgcgccatgg ccatctacaa gcagtcacag cacatgacgg aggttgtgag
     2041 gcgctgcccc caccatgagc gctgctcaga tagcgatggt ctggcccctc ctcagcatct
     2101 tatccgagtg gaaggaaatt tgcgtgtaga gtatttggat gacagaaaca cttttcgaca
     2161 tagtgtggtg gtgccctatg agccgcctga ggttggctct gactgtacca ccatccacta
     2221 caactacatg tgtaacagtt cctgcatggg cggcatgaac cggaggccca tcctcaccat
     2281 catcacactg gaagactcca gtggtaatct actgggacgg aacagctttg aggtgcgtgt
     2341 ttgtgcctgt cctgggagag accggcgcac agaggaagag aatctccgca agaaagggga
     2401 gcctcaccac gagctgcccc cagggagcac taagcgagca ctgcccaaca acaccagctc
     2461 ctctccccag ccaaagaaga aaccactgga tggagaatat ttcacccttc agatccgtgg
     2521 gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc ttggaactca aggatgccca
     2581 ggctgggaag gagccagggg ggagcagggc tcactccagc cacctgaagt ccaaaaaggg
     2641 tcagtctacc tcccgccata aaaaactcat gttcaagaca gaagggcctg actcagactg
     2701 atagtaactc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag
     2761 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2821 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2881 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2941 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3001 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3061 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3121 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3181 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3241 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3301 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3361 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3421 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3481 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3541 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3601 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3661 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3721 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3781 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3841 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3901 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3961 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4021 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4081 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4141 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4201 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4261 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4321 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4381 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4441 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4501 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4561 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4621 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4681 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4741 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4801 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4861 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4921 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4981 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5041 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5101 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5161 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5221 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5281 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5341 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5401 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5461 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5521 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5581 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5641 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5701 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5761 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5821 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5881 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5941 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6001 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6061 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6121 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6181 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6241 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6301 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6361 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6421 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6481 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6541 tcattcagct ccggttccca acg
//
