LOCUS       sequence-249667-        5647 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 5647)
  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 5647)
  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-PGK1-HRE::dLUC
FEATURES             Location/Qualifiers
     source          1..5647
                     /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      250..259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             256..1905
                     /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             1912..1959
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             1963..2082
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2131..2252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2142..2161)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2196..2215
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2446..2803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2446..2637
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2469..2489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2654..2789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2716..2735
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2834..3433
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3458..3506
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3662..3679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3833..4421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3913..3932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4621..5412,5413..5481))
                     /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    5586..5634
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctggcctaac tggccggtac cgagctctgt
      121 cacgtcctgc acgactctag ttgtcacgtc ctgcacgact ctagttgtca cgtcctgcac
      181 gacgctagct cgaggatatc aagatctggc ctcggcggcc aagcttggca atccggtact
      241 gttggtaaag ccaccatgga agatgccaaa aacattaaga agggcccagc gccattctac
      301 ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg ctacgccctg
      361 gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac ctacgccgag
      421 tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct gaatacaaac
      481 catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt gttgggtgcc
      541 ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg cgagctgctg
      601 aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg gctgcaaaag
      661 atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat ggatagcaag
      721 accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt gccacccggc
      781 ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat cgccctgatc
      841 atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca ccgcaccgct
      901 tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat ccccgacacc
      961 gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac gctgggctac
     1021 ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct attcttgcgc
     1081 agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt tagcttcttc
     1141 gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat cgccagcggc
     1201 ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca cctaccaggc
     1261 atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac ccccgaaggg
     1321 gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa ggtggtggac
     1381 ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt ccgtggcccc
     1441 atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat cgacaaggac
     1501 ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt cttcatcgtg
     1561 gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc cgaactggag
     1621 agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct gcccgacgac
     1681 gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac catgaccgag
     1741 aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct gcgcggtggt
     1801 gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc ccgcaagatc
     1861 cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgaattc tgcttgcaag
     1921 aactggttca gtagcttaag ccactttgtg atccacctta acagccacgg cttccctccc
     1981 gaggtggagg agcaggccgc cggcaccctg cccatgagct gcgcccagga gagcggcatg
     2041 gatagacacc ctgctgcttg cgccagcgcc aggatcaacg tctaactgca gtctagagtc
     2101 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2161 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2221 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2281 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2341 gtaaaatcga taaggatccg tttgcgtatt gggcgctctt ccgctgatct gcgcagcacc
     2401 atggcctgaa ataacctctg aaagaggaac ttggttagct accttctgag gcggaaagaa
     2461 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2581 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2641 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2701 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2761 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcg attcttctga
     2821 cactagcgcc accatgaccg agtacaagcc taccgtgcgc ctggccactc gcgatgatgt
     2881 gccccgcgcc gtccgcactc tggccgccgc tttcgccgac taccccgcta cccggcacac
     2941 cgtggacccc gaccggcaca tcgagcgtgt gacagagttg caggagctgt tcctgacccg
     3001 cgtcgggctg gacatcggca aggtgtgggt agccgacgac ggcgcggccg tggccgtgtg
     3061 gactaccccc gagagcgttg aggccggcgc cgtgttcgcc gagatcggcc cccgaatggc
     3121 cgagctgagc ggcagccgcc tggccgccca gcagcaaatg gagggcctgc ttgcccccca
     3181 tcgtcccaag gagcctgcct ggtttctggc cactgtagga gtgagccccg accaccaggg
     3241 caagggcttg ggcagcgccg tcgtgttgcc cggcgtagag gccgccgaac gcgccggtgt
     3301 gcccgccttt ctcgaaacaa gcgcaccaag aaaccttcca ttctacgagc gcctgggctt
     3361 caccgtgacc gccgatgtcg aggtgcccga gggacctagg acctggtgta tgacacgaaa
     3421 acctggcgcc taatgatcta gaaccggtca tggccgcaat aaaatatctt tattttcatt
     3481 acatctgtgt gttggttttt tgtgtgttcg aactagatgc tgtcgaccga tgcccttgag
     3541 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     3601 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     3661 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3901 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3961 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4021 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4081 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4141 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4201 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4261 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4321 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4381 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4501 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4561 aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg
     4621 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
     4681 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
     4741 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
     4801 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
     4861 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
     4921 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
     4981 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
     5041 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
     5101 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
     5161 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
     5221 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
     5281 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
     5341 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
     5401 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
     5461 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
     5521 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
     5581 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     5641 tagtact
//
