LOCUS Exported 5820 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Luciferase reporter construct containing three hypoxia response elements (24-mers) from the Pgk-1 gene.. ACCESSION . VERSION . KEYWORDS HRE-luciferase SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5820) AUTHORS Emerling BM, Weinberg F, Liu JL, Mak TW, Chandel NS TITLE PTEN regulates p300-dependent hypoxia-inducible factor 1 transcriptional activity through Forkhead transcription factor 3a (FOXO3a). JOURNAL Proc Natl Acad Sci U S A. 2008 Feb 19. 105(7):2622-7. PUBMED 18268343 REFERENCE 2 (bases 1 to 5820) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5820 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 85..147 /label=Mini-TK promoter /note="minimal herpes simplex virus (HSV) thymidine kinase promoter" CDS 207..1859 /codon_start=1 /gene="luc" /product="firefly luciferase" /label=luciferase /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL" primer_bind complement(273..291) /label=LucNrev /note="Firefly luciferase, reverse primer" intron 2100..2165 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 2295..2315 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 2740..2874 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3016..3033) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3187..3775) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3267..3286) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3946..4806) /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 4569..4588 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4807..4911) /gene="bla" /label=AmpR promoter rep_origin 4938..5393 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5025..5044) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5235..5256 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 5583..5704 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" ORIGIN 1 tcgagatccg gccccgccca gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc 61 ggggcggcgc ggtccgaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac 121 cgagcgaccc tgcagcgacc cgcttaacag cgtcaacagc gtgccgcaga tctaagtaag 181 cttggcattc cggtactgtt ggtaaaatgg aagacgccaa aaacataaag aaaggcccgg 241 cgccattcta tcctctagag gatggaaccg ctggagagca actgcataag gctatgaaga 301 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtgaacatca 361 cgtacgcgga atacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc 421 tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg 481 tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac 541 gtgaattgct caacagtatg aacatttcgc agcctaccgt agtgtttgtt tccaaaaagg 601 ggttgcaaaa aattttgaac gtgcaaaaaa aattaccaat aatccagaaa attattatca 661 tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc 721 tacctcccgg ttttaatgaa tacgattttg taccagagtc ctttgatcgt gacaaaacaa 781 ttgcactgat aatgaattcc tctggatcta ctgggttacc taagggtgtg gcccttccgc 841 atagaactgc ctgcgtcaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca 901 ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta 961 cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc 1021 tgtttttacg atcccttcag gattacaaaa ttcaaagtgc gttgctagta ccaaccctat 1081 tttcattctt cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa 1141 ttgcttctgg gggcgcacct ctttcgaaag aagtcgggga agcggttgca aaacgcttcc 1201 atcttccagg gatacgacaa ggatatgggc tcactgagac tacatcagct attctgatta 1261 cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga 1321 aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcagagaggc gaattatgtg 1381 tcagaggacc tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga 1441 ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact 1501 tcttcatagt tgaccgcttg aagtctttaa ttaaatacaa aggatatcag gtggcccccg 1561 ctgaattgga atcgatattg ttacaacacc ccaacatctt cgacgcgggc gtggcaggtc 1621 ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga 1681 cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt 1741 tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg 1801 caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagtcc aaattgtaaa 1861 atgtaactgt attcagcgat gacgaaattc ttagctattg taatactgcg atgagtggca 1921 gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg gttgctacgc 1981 ctgaataagt gataataagc ggatgaatgg cagaaattcg ccggatcttt gtgaaggaac 2041 cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa agctctaagg 2101 taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat 2161 tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc tttaatgagg 2221 aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact gctgactctc 2281 aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac tttccttcag 2341 aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct tgctttgcta 2401 tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa aaatattctg 2461 taacctttat aagtaggcat aacagttata atcataacat actgtttttt cttactccac 2521 acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc tttagctttt 2581 taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact agagatcata 2641 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc 2701 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat 2761 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 2821 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgtcga 2881 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact 2941 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca 3001 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 3061 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 3121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 3181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 3241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 3301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 3361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 3421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 3481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 3541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 3601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 3661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 3721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 3781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 3841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 3901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 3961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 4021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 4081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 4141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 4201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 4261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 4321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 4381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 4441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 4501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 4561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 4621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 4681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 4741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 4801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 4861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 4921 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 4981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 5281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 5341 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttgccattc 5401 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg 5461 ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgtggaggtt ttacttgctt 5521 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg 5581 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 5641 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 5701 cttatggtac tgtaactgag ctaacataac ccgggaggta ccgagctctg tcacgtcctg 5761 cacgactcta gttgtcacgt cctgcacgac tctagttgtc acgtcctgca cgacgctagc //