LOCUS       sequence-173934-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia induced mutant p53. Contains the HRE from VEGF sequence (5 
            repeats) upstream of p53 K120R/K164R..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  AUTHORS   Leszczynska KB, Foskolou IP, Abraham AG, Anbalagan S, Tellier C, 
            Haider S, Span PN, O'Neill EE, Buffa FM, Hammond EM
  TITLE     Hypoxia-induced p53 modulates both apoptosis and radiosensitivity 
            via AKT.
  JOURNAL   J Clin Invest. 2015 Jun;125(6):2385-98. doi: 10.1172/JCI80402. Epub 
            2015 May 11.
  PUBMED    25961455
REFERENCE   2  (bases 1 to 7214)
  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"; doi: "10.1172/JCI80402"; 
            journalName: "J Clin Invest"; date: "2015-06"; volume: "125"; issue:
            "6"; pages: "2385-98"
COMMENT     Sequence Label: pGLS3-5xHRE-p53(K120R/K164R)
FEATURES             Location/Qualifiers
     source          1..7214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    129..177
                     /note="synthetic polyadenylation signal"
     misc_feature    191..282
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     231..250
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        519..557
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     554..578
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     595..614
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        595..613
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     638..654
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     639..655
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             823..2004
                     /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
                     GSYGFRLGFLHSGTARSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     RQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(2529..2545)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2529..2545)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2582..2602)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2582..2600)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    2644..2765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2655..2674)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2709..2728
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2941..2964)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3321..3340
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3402..4205
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3465..3484)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4075..4094
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4219..4680
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     primer_bind     complement(4835..4852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5006..5594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5086..5105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5765..6556,6557..6625))
                     /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     6388..6407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6626..6730)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6757..7212
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6844..6863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7054..7075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
       61 ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga
      121 gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
      181 cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat
      241 aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct
      301 tttctcgagc cacagtgcat acgtgggctc caacaggtcc tcttgtcgag ccacagtgca
      361 tacgtgggct ccaacaggtc ctcttgtcga gccacagtgc atacgtgggc tccaacaggt
      421 cctcttgtcg agccacagtg catacgtggg ctccaacagg tcctcttgtc gagccacagt
      481 gcatacgtgg gctccaacag gtcctcttgt cgagatctgg taggcgtgta cggtgggagg
      541 tctatataag cagagctcgt ttagtgaacc gtcagatcac tagaagctcg cgcgtaatac
      601 gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcgacgg tatcgataag
      661 cttgatatcg aattcctcga cggatcccgc tagagccacc gtccagggag caggtagctg
      721 ctgggctccg gggacacttt gcgttcgggc tgggagcgtg ctttccacga cggtgacacg
      781 cttccctgga ttggcagcca gactgccttc cgggtcactg ccatggagga gccgcagtca
      841 gatcctagcg tcgagccccc tctgagtcag gaaacatttt cagacctatg gaaactactt
      901 cctgaaaaca acgttctgtc ccccttgccg tcccaagcaa tggatgattt gatgctgtcc
      961 ccggacgata ttgaacaatg gttcactgaa gacccaggtc cagatgaagc tcccagaatg
     1021 ccagaggctg ctccccccgt ggcccctgca ccagcagctc ctacaccggc ggcccctgca
     1081 ccagccccct cctggcccct gtcatcttct gtcccttccc agaaaaccta ccagggcagc
     1141 tacggtttcc gtctgggctt cttgcattct gggacagcca gatctgtgac ttgcacgtac
     1201 tcccctgccc tcaacaagat gttttgccaa ctggccaaga cctgccctgt gcagctgtgg
     1261 gttgattcca cacccccgcc cggcacccgc gtccgcgcca tggccatcta cagacagtca
     1321 cagcacatga cggaggttgt gaggcgctgc ccccaccatg agcgctgctc agatagcgat
     1381 ggtctggccc ctcctcagca tcttatccga gtggaaggaa atttgcgtgt ggagtatttg
     1441 gatgacagaa acacttttcg acatagtgtg gtggtgccct atgagccgcc tgaggttggc
     1501 tctgactgta ccaccatcca ctacaactac atgtgtaaca gttcctgcat gggcggcatg
     1561 aaccggaggc ccatcctcac catcatcaca ctggaagact ccagtggtaa tctactggga
     1621 cggaacagct ttgaggtgcg tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa
     1681 gagaatctcc gcaagaaagg ggagcctcac cacgagctgc ccccagggag cactaagcga
     1741 gcactgccca acaacaccag ctcctctccc cagccaaaga agaaaccact ggatggagaa
     1801 tatttcaccc ttcagatccg tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag
     1861 gccttggaac tcaaggatgc ccaggctggg aaggagccag gggggagcag ggctcactcc
     1921 agccacctga agtccaaaaa gggtcagtct acctcccgcc ataaaaaact catgttcaag
     1981 acagaagggc ctgactcaga ctgacattct ccacttcttg ttccccactg acagcctccc
     2041 acccccatct ctccctcccc tgccattttg ggttttgggt ctttgaaccc ttgcttgcaa
     2101 taggtgtgcg tcagaagcac ccaggacttc catttgcttt gtcccggggc tccactgaac
     2161 aagttggcct gcactggtgt tttgttgtgg ggaggaggat ggggagtagg acataccagc
     2221 ttagatttta aggtttttac tgtgagggat gtttgggaga tgtaagaaat gttcttgcag
     2281 ttaagggtta gtttacaatc agccacattc taggtagggg cccacttcac cgtactaacc
     2341 agggaagctg tccctcactg ttgaattttc tctaacttca aggcccatat ctgtgaaatg
     2401 ctggcatttg cacctacctc acagagtgca ttgtgagggt taatgaaata atgtacatct
     2461 ggccttgaaa ccacctttta ttacatgggg tctagcggga tccgtcgagg aattcctgca
     2521 gcccggggga tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt
     2581 ccctttagtg agggttaatt gcgcgctaga gtcggggcgg ccggccgctt cgagcagaca
     2641 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2701 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2761 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2821 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cggccgctcg
     2881 agggcccctg caggtcaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     2941 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     3001 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     3061 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     3121 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     3181 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     3241 ttctgggctc agaggctggg aaggggtggg tccgggggsg ggcnnngggg cgggcgcccg
     3301 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     3361 ctcctcttcc tcatctccgg gcctttcgac ctgcagccaa tatgggatcg gccattgaac
     3421 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3481 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3541 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3601 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3661 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3721 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3781 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3841 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3901 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgatgatc
     3961 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4021 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4081 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4141 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4201 tctgagggga tcgatccgct gtaagtctgc agaaattgat gatctattaa acaataaaga
     4261 tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga gaacagagta
     4321 cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg gattagataa
     4381 atgcctgctc tttactgaag gctctttact attgctttat gataatgttt catagttgga
     4441 tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc cactcatgat
     4501 ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc actttgtggt
     4561 tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga gatcagcagc
     4621 ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat tccatcagaa
     4681 gctgactcta gtccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     4741 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     4801 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4861 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4921 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4981 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5041 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5101 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5161 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5221 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5281 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5341 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5401 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5461 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5521 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5581 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5641 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5701 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5761 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5821 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5881 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5941 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6001 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6061 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6121 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6181 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6241 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6301 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6361 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6421 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6481 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6541 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6601 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6661 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6721 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     6781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6901 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6961 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     7021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     7081 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     7141 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     7201 cgcttacaat ttgc
//
