LOCUS       sequence-175207-        5745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia induced mutant PHLDA3. Contains the HRE from VEGF sequence 
            (5 repeats) upstream of PHLDA3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5745)
  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 5745)
  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-PHLDA3(wt)
FEATURES             Location/Qualifiers
     source          1..5745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..31)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     405..424
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             486..1289
                     /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(549..568)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1159..1178
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1303..1764
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     primer_bind     complement(1919..1936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2090..2678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2170..2189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2849..3640,3641..3709))
                     /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     3472..3491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3710..3814)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3841..4296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3928..3947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4138..4159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4427..4475
                     /note="synthetic polyadenylation signal"
     misc_feature    4489..4580
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4529..4548
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4817..4855
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4852..4876
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     4893..4912
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4893..4911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4936..4952
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4937..4953
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5394..5414)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5394..5412)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    5456..5577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5467..5486)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5521..5540
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        join(5718..5745,1..472)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
       61 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
      121 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
      181 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
      241 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
      301 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
      361 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
      421 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
      481 ccaatatggg atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
      541 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
      601 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
      661 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
      721 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
      781 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
      841 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
      901 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
      961 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     1021 cgcgcatgcc cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     1081 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     1141 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     1201 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     1261 tctatcgcct tcttgacgag ttcttctgag gggatcgatc cgctgtaagt ctgcagaaat
     1321 tgatgatcta ttaaacaata aagatgtcca ctaaaatgga agtttttcct gtcatacttt
     1381 gttaagaagg gtgagaacag agtacctaca ttttgaatgg aaggattgga gctacggggg
     1441 tgggggtggg gtgggattag ataaatgcct gctctttact gaaggctctt tactattgct
     1501 ttatgataat gtttcatagt tggatatcat aatttaaaca agcaaaacca aattaagggc
     1561 cagctcattc ctcccactca tgatctatag atctatagat ctctcgtggg atcattgttt
     1621 ttctcttgat tcccactttg tggttctaag tactgtggtt tccaaatgtg tcagtttcat
     1681 agcctgaaga acgagatcag cagcctctgt tccacataca cttcattctc agtattgttt
     1741 tgccaagttc taattccatc agaagctgac tctagtccgt cgaccgatgc ccttgagagc
     1801 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     1861 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     1921 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1981 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2041 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2101 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2161 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2221 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2281 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2341 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2401 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2461 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2521 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2581 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2641 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2701 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2761 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2821 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2881 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2941 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3001 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3061 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3121 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3181 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3241 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3301 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3361 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3421 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3481 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3541 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3601 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3661 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3721 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3781 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3841 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3901 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3961 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4021 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4081 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4141 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4201 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4261 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
     4321 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
     4381 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
     4441 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
     4501 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
     4561 aggtgccaga acatttctct atcgataggt accgagcttt tctcgagcca cagtgcatac
     4621 gtgggctcca acaggtcctc ttgtcgagcc acagtgcata cgtgggctcc aacaggtcct
     4681 cttgtcgagc cacagtgcat acgtgggctc caacaggtcc tcttgtcgag ccacagtgca
     4741 tacgtgggct ccaacaggtc ctcttgtcga gccacagtgc atacgtgggc tccaacaggt
     4801 cctcttgtcg agatctggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     4861 agtgaaccgt cagatcacta gaagctcgcg cgtaatacga ctcactatag ggcgaattgg
     4921 gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgccaccatg acggcggcgg
     4981 cgacggctac cgtgctcaag gagggcgtgc tggagaagcg cagcggcggg ctgctgcagc
     5041 tgtggaagcg gaagcgctgc gtcctcaccg aacgcgggct gcagctcttc gaggccaagg
     5101 gcacgggcgg ccggcccaag gagctcagct tcgcccgcat caaggccgtg gagtgcgtgg
     5161 agagcaccgg gcgccacatc tacttcacgc tggtgaccga agggggcggc gagatcgact
     5221 tccgctgccc cctggaagat cccggctgga acgcccagat caccctaggc ctggtcaagt
     5281 tcaagaacca gcaggccatc cagacagtgc gggcccggca gagcctcggg accgggaccc
     5341 tcgtgtccta atctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     5401 tgagggttaa ttgcgcgcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
     5461 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     5521 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     5581 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     5641 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcggccgct cgagggcccc
     5701 tgcaggtcaa ttctaccggg taggggaggc gcttttccca aggca
//
