LOCUS       sequence-460224-        6563 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses p53 mutant G609C 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  JOURNAL   Unpublished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            "Unpublished"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-G609C
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      247..835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     736..755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     989..1006
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1123..1144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1159..1164,1165..1182,1183..1189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1197..1213
                     /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     1202..1224
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1221..1237
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1221..1237
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1286..1407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1297..1316)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1351..1370
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1581..2375)
                     /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(1692..1711)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2302..2321
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2442..2771)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2456..2591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2510..2529)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2756..2776
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2785..3213)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2895..2916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3107..3126
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3259..3483
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3472..3489
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3509..3527
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3509..3526
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3561..4742)
                     /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(4761..4780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4762..4780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4825..5028)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4854..4874)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5029..5408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5580..5599
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5674..5778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5779..5847,5848..6563,1..76)
                     /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(5997..6016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
       61 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
      121 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      181 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      241 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      301 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      361 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      421 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      481 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      541 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      601 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      661 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      721 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      781 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      841 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      901 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      961 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1021 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1081 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     1141 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     1201 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
     1261 agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact
     1321 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1381 accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc
     1441 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     1501 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     1561 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     1621 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     1681 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     1741 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     1801 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
     1861 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     1921 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     1981 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     2041 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     2101 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     2161 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
     2221 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     2281 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     2341 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     2401 tgatcagatc cgaaaatgga tatacaagct cccgggagct ttttgcaaaa gcctaggcct
     2461 ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc ctcggcctct
     2521 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     2581 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     2641 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     2701 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     2761 aactgacaca cattccacag aattaattcg cgttaaattt ttgttaaatc agctcatttt
     2821 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2881 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2941 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3001 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3061 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3121 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3181 ccgccgcgct taatgcgccg ctacagggcg cgtggggata ccccctagag ccccagctgg
     3241 ttctttccgc ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt
     3301 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
     3361 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
     3421 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
     3481 cagtcgaggc tgatcagcga gctctagcat ttaggtgaca ctatagaata gggccctcta
     3541 gatgcatgct cgagttacta tcagtctgag tcaggccctt ctgtcttgaa catgagtttt
     3601 ttatggcggg aggtagactg accctttttg gacttcaggt ggctggagtg agccctgctc
     3661 ccccctggct ccttcccagc ctgggcatcc ttgagttcca aggcctcatt cagctctcgg
     3721 aacatctcga agcgctcacg cccacggatc tgaagggtga aatattctcc atccagtggt
     3781 ttcttctttg gctggggaga ggagctggtg ttgttgggca gtgctcgctt agtgctccct
     3841 gggggcagct cgtggtgagg ctcccctttc ttgcggagat tctcttcctc tgtgcgccgg
     3901 tctctcccag gacaggcaca aacacgcacc tcaaagctgt tccgtcccag tagattacca
     3961 ctggagtctt ccagtgtgat gatggtgagg atgggcctcc ggttcatgcc gcccatgcag
     4021 gaactgttac acatgtagtt gtagtggatg gtggtacagt cagagccaac ctcaggcggc
     4081 tcatagggca ccaccacact atgtcgaaaa gtgtttctgt catccaaata ctcgacacgc
     4141 aaatttcctt ccactcggat aagatgctga ggaggggcca gaccatcgct atctgagcag
     4201 cgctcatggt gggggcagcg cctcacaacc tccgtcatgt gctgtgactg cttgtagatg
     4261 gccatggcgc ggacgcgggt gccgggcggg ggtgtggaat caacccacag ctgcacaggg
     4321 caggtcttgg ccagttggca aaacatcttg ttgagggcag gggagtacgt gcaagtcaca
     4381 gacttggctg tcccagaatg caagaagccc agacggaaac cgtagctgcc ctggtaggtt
     4441 ttctgggaag ggacagaaga tgacaggggc caggaggggg ctggtgcagg ggccgccggt
     4501 gtaggagctg ctggtgcagg ggccacgggg ggagcagcct ctggcattct gggagcttca
     4561 tctggacctg ggtcttcagt gaaccattgt tcaatatcgt ccggggacag catcaaatca
     4621 tccattgctt gggacggcaa gggggacaga acgttgtttt caggaagtag tttccatagg
     4681 tctgaaaatg tttcctgact cagagggggc tcgacgctag gatctgactg cggctcctcc
     4741 atggcagtaa gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc
     4801 agtgggttct ctagttagcc agagagctct gcttatatag acctcccacc gtacacgcct
     4861 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     4921 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     4981 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     5041 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     5101 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     5161 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     5221 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     5281 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     5341 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     5401 tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct
     5461 aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt aaattttgct
     5521 cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc ttaactatgc
     5581 ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc tcccgatccg
     5641 tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5701 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5761 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5821 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5881 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5941 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6001 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6061 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6121 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6181 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6241 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6301 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6361 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6421 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6481 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6541 cgtatcgtag ttatctacac gac
//
