LOCUS Exported 7452 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS shHIF2-alpha(#2) (HIF2-alpha-#3-pSUPER-retro-GFP/Neo) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7452) AUTHORS Li L, Zhang L, Zhang X, Yan Q, Minamishima YA, Olumi AF, Mao M, Bartz S, Kaelin WG TITLE Hypoxia-inducible factor linked to differential kidney cancer risk seen with type 2A and type 2B VHL mutations. JOURNAL Mol Cell Biol. 2007 Aug . 27(15):5381-92. PUBMED 17526729 REFERENCE 2 (bases 1 to 7452) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7452 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1333..1355 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1371..1387 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind 1421..1437 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 1422..1438 /label=KS primer /note="common sequencing primer, one of multiple similar variants" promoter complement(1511..1725) /label=H1 promoter /note="human H1 RNA promoter" primer_bind complement(1571..1590) /label=H1 /note="Human H1 promoter, forward primer" primer_bind complement(1771..1794) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 2172..2191 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 2268..2277 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2274..2990 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2274..2990 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2319..2340) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2580..2599) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2927..2948 /label=EGFP-C /note="EGFP, forward primer" CDS 3054..3848 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3108..3127) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3718..3737 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3912..3960 /note="synthetic polyadenylation signal" protein_bind 4521..4537 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4545..4575) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4590..4611 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4728..4745) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4899..5487) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4979..4998) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5658..6518) /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 6281..6300 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6519..6623) /gene="bla" /label=AmpR promoter primer_bind 6691..6709 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6747..6769) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6869..6888 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7082..7104 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7232..7251 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 7451..515 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta gatcgatctc tcgaggtcga cggtatcgat 1441 aagcttttcc aaaaaggaga cggaggtgtt ctattttctt gaaatagaac acctccgtct 1501 ccggggatct gtggtctcat acagaactta taagattccc aaatccaaag acatttcacg 1561 tttatggtga tttcccagaa cacatagcga catgcaaata ttgcagggcg ccactcccct 1621 gtccctcaca gccatcttcc tgccagggcg cacgcgcgct gggtgttccc gcctagtgac 1681 actgggcccg cgattccttg gagcgggttg atgacgtcag cgttcgaatt ctaccgggta 1741 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact 1801 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg ccggtaggcg 1861 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag 1921 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg 1981 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg 2041 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg 2101 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc 2161 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc 2221 ctcatctccg ggcctttcga cctgcagccc aagctagctt accggtcgcc accatggtga 2281 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg 2341 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc 2401 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga 2461 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg 2521 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg 2581 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc 2641 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg 2701 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca 2761 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact 2821 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga 2881 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg 2941 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca 3001 gatcagcttg attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg 3061 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3121 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3181 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 3241 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3301 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3361 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3421 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3481 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3541 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 3601 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3661 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 3721 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 3781 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 3841 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 3901 acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg 3961 aatcgatagc gataaggatc ggggatgcat cgataaaata aaagatttta tttagtctcc 4021 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagag aaccatcaga 4081 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc 4141 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac 4201 aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc 4261 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc 4321 ctctgagtga ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt 4381 ggagaacaac attctgaggg taggagtcga atattaagta atcctgactc aattagccac 4441 tgttttgaat ccacatactc caatactcct gaaatagttc attatggaca gcgcagaaag 4501 agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 4561 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 4621 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 4681 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 4741 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4801 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 5101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 5341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5521 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 5581 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 5641 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 5701 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 5761 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 5821 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 5881 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 5941 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 6001 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6061 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6121 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6181 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6241 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6301 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 6361 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 6421 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 6481 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 6541 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 6601 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa 6661 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc 6721 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca 6781 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt 6841 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac 6901 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat 6961 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 7021 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt 7081 tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc 7141 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga 7201 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa 7261 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc 7321 aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc 7381 agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt agtctccaga 7441 aaaagggggg aa //