LOCUS Exported 7799 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mammalian expression. ACCESSION . VERSION . KEYWORDS pcDNA3.1 Flag-HIF1B (#930) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7799) AUTHORS Chen W, Hill H, Christie A, Kim MS, Holloman E, Pavia-Jimenez A, Homayoun F, Ma Y, Patel N, Yell P, Hao G, Yousuf Q, Joyce A, Pedrosa I, Geiger H, Zhang H, Chang J, Gardner KH, Bruick RK, Reeves C, Hwang TH, Courtney K, Frenkel E, Sun X, Zojwalla N, Wong T, Rizzi JP, Wallace EM, Josey JA, Xie Y, Xie XJ, Kapur P, McKay RM, Brugarolas J TITLE Targeting renal cell carcinoma with a HIF-2 antagonist. JOURNAL Nature. 2016 Nov 3;539(7627):112-117. doi: 10.1038/nature19796. Epub 2016 Sep 5. PUBMED 27595394 REFERENCE 2 (bases 1 to 7799) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7799 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 683..692 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 692..715 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(3156..3173) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3162..3386 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3432..3860 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3519..3538) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3729..3750 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3869..3889) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3874..4203 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4054..4189 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4116..4135 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4270..5064 /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(4324..4343) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4934..4953 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5238..5359 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5275..5294) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5329..5348 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5408..5424) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5408..5424) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5421..5443) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5432..5448 /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(5456..5486) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5501..5522 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5639..5656) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5810..6398) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5890..5909) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6569..7429) /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 7192..7211 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7430..7534) /gene="bla" /label=AmpR promoter primer_bind complement(7609..7628) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgttt aaacttaagc ttgccaccat ggattacaag gatgacgacg ataaggcggc 721 gactactgcc aaccccgaaa tgacatcaga tgtaccatca ctgggtccag ccattgcctc 781 tggaaactct ggacctggaa ttcaaggtgg aggagccatt gtccagaggg ctattaagcg 841 gcgaccaggg ctggattttg atgatgatgg agaagggaac agtaaatttt tgaggtgtga 901 tgatgatcag atgtctaacg ataaggagcg gtttgccagg tcggatgatg agcagagctc 961 tgcggataaa gagagacttg ccagggaaaa tcacagtgaa attgaacggc ggcgacggaa 1021 caagatgaca gcctacatca cagaactgtc agatatggta cccacctgta gtgccctggc 1081 tcgaaaacca gacaagctaa ccatcttacg catggcagtt tctcacatga agtccttgcg 1141 gggaactggc aacacatcca ctgatggctc ctataagccg tctttcctca ctgatcagga 1201 actgaaacat ttgatcttgg aggcagcaga tggctttctg tttattgtct catgtgagac 1261 aggcagggtg gtgtatgtgt ctgactccgt gactcctgtt ttgaaccagc cacagtctga 1321 atggtttggc agcacactct atgatcaggt gcacccagat gatgtggata aacttcgtga 1381 gcagctttcc acttcagaaa atgccctgac agggcgtatc ctggatctaa agactggaac 1441 agtgaaaaag gaaggtcagc agtcttccat gagaatgtgt atgggctcaa ggagatcgtt 1501 tatttgccga atgaggtgtg gcagtagctc tgtggaccca gtttctgtga ataggctgag 1561 ctttgtgagg aacagatgca ggaatggact tggctctgta aaggatgggg aacctcactt 1621 cgtggtggtc cactgcacag gctacatcaa ggcctggccc ccagcaggtg tttccctccc 1681 agatgatgac ccagaggctg gccagggaag caagttttgc ctagtggcca ttggcagatt 1741 gcaggtaact agttctccca actgtacaga catgagtaat gtttgtcaac caacagagtt 1801 catctcccga cacaacattg agggtatctt cacttttgtg gatcaccgct gtgtggctac 1861 tgttggctac cagccacagg aactcttagg aaagaatatt gtagaattct gtcatcctga 1921 agaccagcag cttctaagag acagcttcca acaggtagtg aaattaaaag gccaagtgct 1981 gtctgtcatg ttccggttcc ggtctaagaa ccaagaatgg ctctggatga gaaccagctc 2041 ctttactttc cagaaccctt actcagatga aattgagtac atcatctgta ccaacaccaa 2101 tgtgaagaac tctagccaag aaccacggcc tacactctcc aacacaatcc agaggccaca 2161 actaggtccc acagctaatt tacccctgga gatgggctca ggacagctgg cacccaggca 2221 gcagcaacag caaacagaat tggacatggt accaggaaga gatggactgg ccagctacaa 2281 tcattcccag gtggttcagc ctgtgacaac cacaggacca gaacacagca agccccttga 2341 gaagtcagat ggtttatttg cccaggatag agatccaaga ttttcagaaa tctatcacaa 2401 catcaatgcg gatcagagta aaggcatctc ctccagcact gtccctgcca cccaacagct 2461 attctcccag ggcaacacat tccctcctac cccccggccg gcagagaatt tcaggaatag 2521 tggcctagcc cctcctgtaa ccattgtcca gccatcagct tctgcaggac agatgttggc 2581 ccagatttcc cgccactcca accccaccca aggagcaacc ccaacttgga cccctactac 2641 ccgctcaggc ttttctgccc agcaggtggc tacccaggct actgctaaga ctcgtacttc 2701 ccagtttggt gtgggcagct ttcagactcc atcctccttc agctccatgt ccctccctgg 2761 tgccccaact gcatcgcctg gtgctgctgc ctaccctagt ctcaccaatc gtggatctaa 2821 ctttgctcct gagactggac agactgcagg acaattccag acacggacag cagagggtgt 2881 gggtgtctgg ccacagtggc agggccagca gcctcatcat cgttcaagtt ctagtgagca 2941 acatgttcaa caaccgccag cacagcaacc tggccagcct gaggtcttcc aggagatgct 3001 gtccatgctg ggagatcaga gcaacagcta caacaatgaa gaattccctg atctaactat 3061 gtttcccccc ttttcagaat gataagaatt ctgcagatat ccagcacagt ggcggccgct 3121 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 3181 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 3241 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 3301 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 3361 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 3421 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 3481 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 3541 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 3601 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 3661 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 3721 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 3781 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 3841 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4021 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 4081 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 4141 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 4201 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 4261 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 4321 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 4381 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 4441 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 4501 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 4561 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 4621 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 4681 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 4741 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 4801 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 4861 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 4921 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 4981 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 5041 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 5101 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 5161 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 5221 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5341 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 5401 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5461 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5521 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5581 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 5641 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 5701 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 5761 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 5821 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 5881 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 5941 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 6001 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 6061 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 6121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 6181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6241 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 6361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 6781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 6841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 7021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 7081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 7141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 7201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 7261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7561 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc 7621 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta 7681 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa 7741 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt //