LOCUS       sequence-364702-        7314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7314)
  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 7314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol. 2005 Jan . 25(1):162-71."
COMMENT     Sequence Label: pcDNA3-HA2-Keap1
FEATURES             Location/Qualifiers
     source          1..7314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2033..2052)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2224..2603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2604..2807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2758..2778
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2852..2871
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2852..2870
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2916..2942
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2922..2948
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(4856..4874)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4857..4874)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4894..4911)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4900..5124
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5170..5598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5257..5276)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5467..5488
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5607..5627)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5612..5941
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5792..5927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5854..5873
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6008..6802
                     /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(6062..6081)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6672..6691
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6976..7097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7013..7032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7067..7086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7146..7162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7146..7162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7159..7181)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7170..7186
                     /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(join(7194..7200,7201..7218,7219..7224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7239..7260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
     2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gasacccaag
     2881 ctatgtaccc atacgakgtt cctgastawg sgggctatcc ctatgacgtc ccggactatg
     2941 caagcttggt accgttaaca tggatcccca tgcagccaga tcccaggcct agcggggctg
     3001 gggcctgctg ccgattcctg cccctgcagt cacagtgccc tgagggggca ggggacgcgg
     3061 tgatgtacgc ctccactgag tgcaaggcgg aggtgacgcc ctcccagcat ggcaaccgca
     3121 ccttcagcta caccctggag gatcatacca agcaggcctt tggcatcatg aacgagctgc
     3181 ggctcagcca gcagctgtgt gacgtcacac tgcaggtcaa gtaccaggat gcaccggccg
     3241 cccagttcat ggcccacaag gtggtgctgg cctcatccag ccctgtcttc aaggccatgt
     3301 tcaccaacgg gctgcgggag cagggcatgg aggtggtgtc cattgagggt atccacccca
     3361 aggtcatgga gcgcctcatt gaattcgcct acacggcctc catctccatg ggcgagaagt
     3421 gtgtcctcca cgtcatgaac ggtgctgtca tgtaccagat cgacagcgtt gtccgtgcct
     3481 gcagtgactt cctggtgcag cagctggacc ccagcaatgc catcggcatc gccaacttcg
     3541 ctgagcagat tggctgtgtg gagttgcacc agcgtgcccg ggagtacatc tacatgcatt
     3601 ttggggaggt ggccaagcaa gaggagttct tcaacctgtc ccactgccaa ctggtgaccc
     3661 tcatcagccg ggacgacctg aacgtgcgct gcgagtccga ggtcttccac gcctgcatca
     3721 actgggtcaa gtacgactgc gaacagcgac ggttctacgt ccaggcgctg ctgcgggccg
     3781 tgcgctgcca ctcgttgacg ccgaacttcc tgcagatgca gctgcagaag tgcgagatcc
     3841 tgcagtccga ctcccgctgc aaggactacc tggtcaagat cttcgaggag ctcaccctgc
     3901 acaagcccac gcaggtgatg ccctgccggg cgcccaaggt gggccgcctg atctacaccg
     3961 cgggcggcta cttccgacag tcgctcagct acctggaggc ttacaacccc agtgacggca
     4021 cctggctccg gttggcggac ctgcaggtgc cgcggagcgg cctggccggc tgcgtggtgg
     4081 gcgggctgtt gtacgccgtg ggcggcagga acaactcgcc cgacggcaac accgactcca
     4141 gcgccctgga ctgttacaac cccatgacca atcagtggtc gccctgcgcc cccatgagcg
     4201 tgccccgtaa ccgcatcggg gtgggggtca tcgatggcca catctatgcc gtcggcggct
     4261 cccacggctg catccaccac aacagtgtgg agaggtatga gccagagcgg gatgagtggc
     4321 acttggtggc cccaatgctg acacgaagga tcggggtggg cgtggctgtc ctcaatcgtc
     4381 tcctttatgc cgtggggggc tttgacggga caaaccgcct taattcagct gagtgttact
     4441 acccagagag gaacgagtgg cgaatgatca cagcaatgaa caccatccga agcggggcag
     4501 gcgtctgcgt cctgcacaac tgtatctatg ctgctggggg ctatgatggt caggaccagc
     4561 tgaacagcgt ggagcgctac gatgtggaaa cagagacgtg gactttcgta gcccccatga
     4621 agcaccggcg aagtgccctg gggatcactg tccaccaggg gagaatctac gtccttggag
     4681 gctatgatgg tcacacgttc ctggacagtg tggagtgtta cgacccagat acagacacct
     4741 ggagcgaggt gacccgaatg acatcgggcc ggagtggggt gggcgtggct gtcaccatgg
     4801 agccctgccg gaagcagatt gaccagcaga actgtacctg tctagagggc cctattctat
     4861 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     4921 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4981 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     5041 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     5101 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     5161 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5221 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5281 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5341 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5401 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5461 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5521 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5581 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5641 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5701 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5761 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5821 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5881 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5941 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     6001 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     6061 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     6121 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6181 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     6241 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6301 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     6361 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6421 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6481 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     6541 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6601 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6661 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6721 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6781 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6841 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6901 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6961 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     7021 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     7081 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     7141 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7201 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7261 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctg
//
