LOCUS Exported 7972 bp ds-DNA circular SYN 05-JAN-2023 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-EGFP-C4-Nrf2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7972) AUTHORS Furukawa M, Xiong Y TITLE BTB protein Keap1 targets antioxidant transcription factor Nrf2 for ubiquitination by the Cullin 3-Roc1 ligase. JOURNAL Mol Cell Biol. 2005 Jan . 25(1):162-71. PUBMED 15601839 REFERENCE 2 (bases 1 to 7972) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 5, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7972 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 51..430 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 431..634 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 585..605 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 679..698 /label=T7 /note="T7 promoter, forward primer" promoter 679..697 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 723..732 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 729..1445 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 729..1445 /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(774..795) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1035..1054) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1382..1403 /label=EGFP-C /note="EGFP, forward primer" CDS 1464..1493 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" promoter complement(3341..3359) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3342..3359) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3379..3396) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3385..3609 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3655..4083 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3742..3761) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3952..3973 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4092..4112) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4097..4426 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4277..4412 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4339..4358 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4493..5287 /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(4547..4566) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5157..5176 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5461..5582 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5498..5517) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5552..5571 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5631..5647) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5631..5647) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5644..5666) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5655..5671 /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(5679..5709) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5724..5745 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5862..5879) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6033..6621) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6113..6132) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6792..7652) /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 7415..7434 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7653..7757) /gene="bla" /label=AmpR promoter primer_bind complement(7832..7851) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt 61 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 601 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc 661 ttactggctt atcgaaatta atacgactca ctatagggag acccaagcta gcgctaccgg 721 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg 781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg 841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct 901 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc 961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca 1021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg 1081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc 1141 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc 1201 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc 1261 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg 1321 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc 1381 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt 1441 acaagtactc agatctggaa ttagagcaaa agctcatctc agaggaagat ctggaattca 1501 ggatcctcat gatggacttg gagctgccgc cgccgggact cccgtcccag caggacatgg 1561 atttgattga catactttgg aggcaagata tagatcttgg agtaagtcga gaagtatttg 1621 acttcagtca gcgacggaaa gagtatgagc tggaaaaaca gaaaaaactt gaaaaggaaa 1681 gacaagaaca actccaaaag gagcaagaga aagccttttt cgctcagtta caactagatg 1741 aagagacagg tgaatttctc ccaattcagc cagcccagca catccagtca gaaaccagtg 1801 gatctgccaa ctactcccag gttgcccaca ttcccaaatc agatgctttg tactttgatg 1861 actgcatgca gcttttggcg cagacattcc cgtttgtaga tgacaatgag gtttcttcgg 1921 ctacgtttca gtcacttgtt cctgatattc ccggtcacat cgagagccca gtcttcattg 1981 ctactaatca ggctcagtca cctgaaactt ctgttgctca ggtagcccct gttgatttag 2041 acggtatgca acaggacatt gagcaagttt gggaggagct attatccatt cctgagttac 2101 agtgtcttaa tattgaaaat gacaagctgg ttgagactac catggttcca agtccagaag 2161 ccaaactgac agaagttgac aattatcatt tttactcatc tataccctca atggaaaaag 2221 aagtaggtaa ctgtagtcca cattttctta atgcttttga ggattccttc agcagcatcc 2281 tctccacaga agaccccaac cagttgacag tgaactcatt aaattcagat gccacagtca 2341 acacagattt tggtgatgaa ttttattctg ctttcatagc tgagcccagt atcagcaaca 2401 gcatgccctc acctgctact ttaagccatt cactctctga acttctaaat gggcccattg 2461 atgtttctga tctatcactt tgcaaagctt tcaaccaaaa ccaccctgaa agcacagcag 2521 aattcaatga ttctgactcc ggcatttcac taaacacaag tcccagtgtg gcatcaccag 2581 aacactcagt ggaatcttcc agctatggag acacactact tggcctcagt gattctgaag 2641 tggaagagct agatagtgcc cctggaagtg tcaaacagaa tggtcctaaa acaccagtac 2701 attcttctgg ggatatggta caacccttgt caccatctca ggggcagagc actcacgtgc 2761 atgatgccca atgtgagaac acaccagaga aagaattgcc tgtaagtcct ggtcatcgga 2821 aaaccccatt cacaaaagac aaacattcaa gccgcttgga ggctcatctc acaagagatg 2881 aacttagggc aaaagctctc catatcccat tccctgtaga aaaaatcatt aacctccctg 2941 ttgttgactt caacgaaatg atgtccaaag agcagttcaa tgaagctcaa cttgcattaa 3001 ttcgggatat acgtaggagg ggtaagaata aagtggctgc tcagaattgc agaaaaagaa 3061 aactggaaaa tatagtagaa ctagagcaag atttagatca tttgaaagat gaaaaagaaa 3121 aattgctcaa agaaaaagga gaaaatgaca aaagccttca cctactgaaa aaacaactca 3181 gcaccttata tctcgaagtt ttcagcatgc tacgtgatga agatggaaaa ccttattctc 3241 ctagtgaata ctccctgcag caaacaagag atggcaatgt tttccttgtt cccaaaagta 3301 agaagccaga tgttaagaaa aactatctag agggccctat tctatagtgt cacctaaatg 3361 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 3421 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 3481 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 3541 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 3601 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 3661 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3721 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3781 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 3841 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3901 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 3961 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 4021 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 4081 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 4141 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 4201 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 4261 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 4321 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 4381 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 4441 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 4501 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 4561 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 4621 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 4681 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 4741 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 4801 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 4861 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 4921 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 4981 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 5041 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 5101 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 5161 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 5221 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 5281 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 5341 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 5401 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 5461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 5521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 5581 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 5641 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 5701 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 5761 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 5821 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 5881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 5941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 6001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 6061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 6121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 6181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 6241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 6301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 6361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 6421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 6481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 6601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 6661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 6781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 6841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 6901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 6961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 7021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 7081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 7141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 7201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 7261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 7321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 7381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 7441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 7501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 7561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 7621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 7681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 7741 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga 7801 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 7861 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt 7921 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt ag //