LOCUS       sequence-424651-        7325 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 7325)
  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 7325)
  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 S3H4
FEATURES             Location/Qualifiers
     source          1..7325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..32
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(151..171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        156..485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      336..471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     398..417
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             552..1346
                     /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(606..625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1216..1235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1520..1641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1557..1576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1611..1630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1690..1706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1690..1706)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1703..1725)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1714..1730
                     /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(1738..1744,1745..1762,1763..1768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1783..1804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1921..1938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2103..2691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2183..2202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2862..3653,3654..3722))
                     /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     3485..3504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3723..3827)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3902..3921)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4093..4472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4473..4676
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4627..4647
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4721..4740
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4721..4739
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4755..4781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4785..4811
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4791..4817
                     /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(6725..6743)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6726..6743)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6763..6780)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6769..6993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7039..7325,1..142)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7126..7145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
       61 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      121 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      421 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
      541 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      601 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      661 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      721 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      781 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      841 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      901 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      961 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1021 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1081 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1141 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1201 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1261 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1321 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1381 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1441 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1501 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     1561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1621 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     1681 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1741 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1801 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1861 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2041 aaaaggccag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2281 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2521 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3841 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     3901 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     3961 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     4021 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     4081 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     4141 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4201 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4261 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4321 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4381 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4441 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     4501 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     4561 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     4621 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
     4681 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
     4741 agacccaagc tatgtaccca tacgatgttc ctgactatgc gggctatccc tatgacgtcc
     4801 cggactatgc aagcttggta ccgttaacat ggatccccat gcagccagat cccaggccta
     4861 gcggggctgg ggcctgctgc cgattcctgc ccctgcagtc acagtgccct gagggggcag
     4921 gggacgcggt gatgtacgcc tccactgagt gcaaggcgga ggtgacgccc tcccagcatg
     4981 gcaaccgcac cttcagctac accctggagg atcataccaa gcaggccttt ggcatcatga
     5041 acgagctgcg gctcagccag cagctgtgtg acgtcacact gcaggtcaag taccaggatg
     5101 caccggccgc ccagttcatg gcccacaagg tggtgctggc ctcatccagc cctgtcttca
     5161 aggccatgtt caccaacggg ctgcgggagc agggcatgga ggtggctagc gctgaggcta
     5221 tccaccccaa ggtcatggag cgcctcattg aattcgccta cacggcctcc atctccatgg
     5281 gcgagaagtg tgtcctccac gtcatgaacg gtgccgcggc gtaccagatc gacagcgttg
     5341 tccgtgcctg cagtgacttc ctggtgcagc agctggaccc cagcaatgcc atcggcatcg
     5401 ccaacttcgc tgagcagatt ggctgtgtgg agttgcacca gcgtgcccgg gagtacatct
     5461 acatgcattt tggggaggtg gccaagcaag aggagttctt caacctgtcc cactgccaac
     5521 tggtgaccct catcagccgg gacgacctga acgtgcgctg cgagtccgag gtcttccacg
     5581 cctgcatcaa ctgggtcaag tacgactgcg aacagcgacg gttctacgtc caggcgctgc
     5641 tgcgggccgt gcgctgccac tcgttgacgc cgaacttcct gcagatgcag ctgcagaagt
     5701 gcgagatcct gcagtccgac tcccgctgca aggactacct ggtcaagatc ttcgaggagc
     5761 tcaccctgca caagcccacg caggtgatgc cctgccgggc gcccaaggtg ggccgcctga
     5821 tctacaccgc gggcggctac ttccgacagt cgctcagcta cctggaggct tacaacccca
     5881 gtgacggcac ctggctccgg ttggcggacc tgcaggtgcc gcggagcggc ctggccggct
     5941 gcgtggtggg cgggctgttg tacgccgtgg gcggcaggaa caactcgccc gacggcaaca
     6001 ccgactccag cgccctggac tgttacaacc ccatgaccaa tcagtggtcg ccctgcgccc
     6061 ccatgagcgt gccccgtaac cgcatcgggg tgggggtcat cgatggccac atctatgccg
     6121 tcggcggctc ccacggctgc atccaccaca acagtgtgga gaggtatgag ccagagcggg
     6181 atgagtggca cttggtggcc ccaatgctga cacgaaggat cggggtgggc gtggctgtcc
     6241 tcaatcgtct cctttatgcc gtggggggct ttgacgggac aaaccgcctt aattcagctg
     6301 agtgttacta cccagagagg aacgagtggc gaatgatcac agcaatgaac accatccgaa
     6361 gcggggcagg cgtctgcgtc ctgcacaact gtatctatgc tgctgggggc tatgatggtc
     6421 aggaccagct gaacagcgtg gagcgctacg atgtggaaac agagacgtgg actttcgtag
     6481 cccccatgaa gcaccggcga agtgccctgg ggatcactgt ccaccagggg agaatctacg
     6541 tccttggagg ctatgatggt cacacgttcc tggacagtgt ggagtgttac gacccagata
     6601 cagacacctg gagcgaggtg acccgaatga catcgggccg gagtggggtg ggcgtggctg
     6661 tcaccatgga gccctgccgg aagcagattg accagcagaa ctgtacctgt ctagagggcc
     6721 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     6781 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     6841 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     6901 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     6961 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     7021 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     7081 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     7141 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     7201 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     7261 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     7321 ccacg
//
