pcDNA3-EGFP-C4-Nrf2

LOCUS       pcDNA3-EGFP-C4-Nrf2                7972 bp ds-DNA     circular SYN 22-AUG-2018
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 Aug 22, 2018 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
                     /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
//