LOCUS       sequence-152114-        7285 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 7285)
  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 7285)
  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-T7-Keap1
FEATURES             Location/Qualifiers
     source          1..7285
                     /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             2893..2925
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     promoter        complement(4827..4845)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4828..4845)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4865..4882)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4871..5095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5141..5569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5228..5247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5438..5459
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5578..5598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5583..5912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5763..5898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5825..5844
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5979..6773
                     /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(6033..6052)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6643..6662
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6947..7068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6984..7003)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7038..7057
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7117..7133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7117..7133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7130..7152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7141..7157
                     /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(7165..7171,7172..7189,7190..7195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7210..7231
                     /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 gagacccaag
     2881 cttggtacca ccatggctag catgactggt ggacagcaaa tgggtgaatt caggatcccc
     2941 atgcagccag atcccaggcc tagcggggct ggggcctgct gccgattcct gcccctgcag
     3001 tcacagtgcc ctgagggggc aggggacgcg gtgatgtacg cctccactga gtgcaaggcg
     3061 gaggtgacgc cctcccagca tggcaaccgc accttcagct acaccctgga ggatcatacc
     3121 aagcaggcct ttggcatcat gaacgagctg cggctcagcc agcagctgtg tgacgtcaca
     3181 ctgcaggtca agtaccagga tgcaccggcc gcccagttca tggcccacaa ggtggtgctg
     3241 gcctcatcca gccctgtctt caaggccatg ttcaccaacg ggctgcggga gcagggcatg
     3301 gaggtggtgt ccattgaggg tatccacccc aaggtcatgg agcgcctcat tgaattcgcc
     3361 tacacggcct ccatctccat gggcgagaag tgtgtcctcc acgtcatgaa cggtgctgtc
     3421 atgtaccaga tcgacagcgt tgtccgtgcc tgcagtgact tcctggtgca gcagctggac
     3481 cccagcaatg ccatcggcat cgccaacttc gctgagcaga ttggctgtgt ggagttgcac
     3541 cagcgtgccc gggagtacat ctacatgcat tttggggagg tggccaagca agaggagttc
     3601 ttcaacctgt cccactgcca actggtgacc ctcatcagcc gggacgacct gaacgtgcgc
     3661 tgcgagtccg aggtcttcca cgcctgcatc aactgggtca agtacgactg cgaacagcga
     3721 cggttctacg tccaggcgct gctgcgggcc gtgcgctgcc actcgttgac gccgaacttc
     3781 ctgcagatgc agctgcagaa gtgcgagatc ctgcagtccg actcccgctg caaggactac
     3841 ctggtcaaga tcttcgagga gctcaccctg cacaagccca cgcaggtgat gccctgccgg
     3901 gcgcccaagg tgggccgcct gatctacacc gcgggcggct acttccgaca gtcgctcagc
     3961 tacctggagg cttacaaccc cagtgacggc acctggctcc ggttggcgga cctgcaggtg
     4021 ccgcggagcg gcctggccgg ctgcgtggtg ggcgggctgt tgtacgccgt gggcggcagg
     4081 aacaactcgc ccgacggcaa caccgactcc agcgccctgg actgttacaa ccccatgacc
     4141 aatcagtggt cgccctgcgc ccccatgagc gtgccccgta accgcatcgg ggtgggggtc
     4201 atcgatggcc acatctatgc cgtcggcggc tcccacggct gcatccacca caacagtgtg
     4261 gagaggtatg agccagagcg ggatgagtgg cacttggtgg ccccaatgct gacacgaagg
     4321 atcggggtgg gcgtggctgt cctcaatcgt ctcctttatg ccgtgggggg ctttgacggg
     4381 acaaaccgcc ttaattcagc tgagtgttac tacccagaga ggaacgagtg gcgaatgatc
     4441 acagcaatga acaccatccg aagcggggca ggcgtctgcg tcctgcacaa ctgtatctat
     4501 gctgctgggg gctatgatgg tcaggaccag ctgaacagcg tggagcgcta cgatgtggaa
     4561 acagagacgt ggactttcgt agcccccatg aagcaccggc gaagtgccct ggggatcact
     4621 gtccaccagg ggagaatcta cgtccttgga ggctatgatg gtcacacgtt cctggacagt
     4681 gtggagtgtt acgacccaga tacagacacc tggagcgagg tgacccgaat gacatcgggc
     4741 cggagtgggg tgggcgtggc tgtcaccatg gagccctgcc ggaagcagat tgaccagcag
     4801 aactgtacct gtctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     4861 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4921 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4981 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     5041 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     5101 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     5161 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5221 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5281 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5341 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5401 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5461 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5521 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5641 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5701 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5761 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5821 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5881 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5941 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     6001 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6061 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6121 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     6181 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6241 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6301 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6361 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6421 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6481 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6541 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6601 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6661 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6721 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6781 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6841 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6901 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7081 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7261 gtcgggaaac ctgtcgtgcc agctg
//
