LOCUS       sequence-249669-        5779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia response element real-time luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5779)
  AUTHORS   Walton ZE, Patel CH, Brooks RC, Yu Y, Ibrahim-Hashim A, Riddle M, 
            Porcu A, Jiang T, Ecker BL, Tameire F, Koumenis C, Weeraratna AT, 
            Welsh DK, Gillies R, Alwine JC, Zhang L, Powell JD, Dang CV
  TITLE     Acid Suspends the Circadian Clock in Hypoxia through Inhibition of 
            mTOR.
  JOURNAL   Cell. 2018 Jun 28;174(1):72-87.e32. doi: 10.1016/j.cell.2018.05.009.
            Epub 2018 May 31.
  PUBMED    29861175
REFERENCE   2  (bases 1 to 5779)
  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: 
            "10.1016/j.cell.2018.05.009"; journalName: "Cell"; date: 
            "2018-06-28- 28"; volume: "174"; issue: "1"; pages: "72-87.e32"
COMMENT     Sequence Label: pGL4.22-VEGF-HRE::dLUC
FEATURES             Location/Qualifiers
     source          1..5779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      382..391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             388..2037
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2044..2091
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             2095..2214
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2263..2384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2274..2293)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2328..2347
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2578..2935
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2578..2769
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2601..2621)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2786..2921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2848..2867
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2966..3565
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3590..3638
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3794..3811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3965..4553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4045..4064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4753..5544,5545..5613))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5718..5766
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctggcctaac tggccggtac ctgagctcgc
      121 tagcctcgag ccacagtgca tacgtgggct ccaacaggtc ctcttgtcga gccacagtgc
      181 atacgtgggc tccaacaggt cctcttgtcg agccacagtg catacgtggg ctccaacagg
      241 tcctcttgtc gagccacagt gcatacgtgg gctccaacag gtcctcttgt cgagccacag
      301 tgcatacgtg ggctccaaca ggtcctcttg tcgagatctg gcctcggcgg ccaagcttgg
      361 caatccggta ctgttggtaa agccaccatg gaagatgcca aaaacattaa gaagggccca
      421 gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag
      481 cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt
      541 acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg
      601 ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc
      661 gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag
      721 cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa
      781 gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc
      841 atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat
      901 ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc
      961 atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg
     1021 caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc
     1081 atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc
     1141 acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag
     1201 ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta
     1261 tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag
     1321 atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc
     1381 cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc
     1441 acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct
     1501 aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc
     1561 gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc
     1621 atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac
     1681 ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca
     1741 gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc
     1801 ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa
     1861 accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag
     1921 ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac
     1981 gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgaat
     2041 tctgcttgca agaactggtt cagtagctta agccactttg tgatccacct taacagccac
     2101 ggcttccctc ccgaggtgga ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag
     2161 gagagcggca tggatagaca ccctgctgct tgcgccagcg ccaggatcaa cgtctaactg
     2221 cagtctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     2281 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2341 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2401 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     2461 tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc ttccgctgat
     2521 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag ctaccttctg
     2581 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2761 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2821 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     2881 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2941 cgattcttct gacactagcg ccaccatgac cgagtacaag cctaccgtgc gcctggccac
     3001 tcgcgatgat gtgccccgcg ccgtccgcac tctggccgcc gctttcgccg actaccccgc
     3061 tacccggcac accgtggacc ccgaccggca catcgagcgt gtgacagagt tgcaggagct
     3121 gttcctgacc cgcgtcgggc tggacatcgg caaggtgtgg gtagccgacg acggcgcggc
     3181 cgtggccgtg tggactaccc ccgagagcgt tgaggccggc gccgtgttcg ccgagatcgg
     3241 cccccgaatg gccgagctga gcggcagccg cctggccgcc cagcagcaaa tggagggcct
     3301 gcttgccccc catcgtccca aggagcctgc ctggtttctg gccactgtag gagtgagccc
     3361 cgaccaccag ggcaagggct tgggcagcgc cgtcgtgttg cccggcgtag aggccgccga
     3421 acgcgccggt gtgcccgcct ttctcgaaac aagcgcacca agaaaccttc cattctacga
     3481 gcgcctgggc ttcaccgtga ccgccgatgt cgaggtgccc gagggaccta ggacctggtg
     3541 tatgacacga aaacctggcg cctaatgatc tagaaccggt catggccgca ataaaatatc
     3601 tttattttca ttacatctgt gtgttggttt tttgtgtgtt cgaactagat gctgtcgacc
     3661 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     3721 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     3781 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3841 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3901 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3961 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4021 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4081 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4141 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4201 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4261 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4321 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4381 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4441 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4501 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4561 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4621 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4681 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc aaatgctaaa
     4741 ccactgcagt ggttaccagt gcttgatcag tgaggcaccg atctcagcga tctgcctatt
     4801 tcgttcgtcc atagtggcct gactccccgt cgtgtagatc actacgattc gtgagggctt
     4861 accatcaggc cccagcgcag caatgatgcc gcgagagccg cgttcaccgg cccccgattt
     4921 gtcagcaatg aaccagccag cagggagggc cgagcgaaga agtggtcctg ctactttgtc
     4981 cgcctccatc cagtctatga gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag
     5041 tagtttccga agagttgtgg ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg
     5101 tatggcttcg ttcaactctg gttcccagcg gtcaagccgg gtcacatgat cacccatatt
     5161 atgaagaaat gcagtcagct ccttagggcc tccgatcgtt gtcagaagta agttggccgc
     5221 ggtgttgtcg ctcatggtaa tggcagcact acacaattct cttaccgtca tgccatccgt
     5281 aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg
     5341 gcgaccaagc tgctcttgcc cggcgtctat acgggacaac accgcgccac atagcagtac
     5401 tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc
     5461 gctattgaga tccagttcga tatagcccac tcttgcaccc agttgatctt cagcatcttt
     5521 tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg
     5581 aatgagtgcg acacgaaaat gttggatgct catactcgtc ctttttcaat attattgaag
     5641 catttatcag ggttactagt acgtctctca aggataagta agtaatatta aggtacggga
     5701 ggtattggac aggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     5761 tgtgtgaatc gatagtact
//
