LOCUS       sequence-156377-        6506 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 6506)
  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 6506)
  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: pDsRed2-Keap1
FEATURES             Location/Qualifiers
     source          1..6506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1294
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(800..820)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1212..1235
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3301..3422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3338..3357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3392..3411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3429..3884)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3566..3587)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3778..3797
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3911..4015
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4017..4374
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4017..4208
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4040..4060)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4225..4360
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4409..5203
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4409..5203
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4463..4482)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5073..5092
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5391..5410)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5435..5482
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5811..6399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6300..6319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggcctcctc cgagaacgtc atcaccgagt tcatgcgctt
      661 caaggtgcgc atggagggca ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      721 gggccgcccc tacgagggcc acaacaccgt gaagctgaag gtgaccaagg gcggccccct
      781 gcccttcgcc tgggacatcc tgtcccccca gttccagtac ggctccaagg tgtacgtgaa
      841 gcaccccgcc gacatccccg actacaagaa gctgtccttc cccgagggct tcaagtggga
      901 gcgcgtgatg aacttcgagg acggcggcgt ggcgaccgtg acccaggact cctccctgca
      961 ggacggctgc ttcatctaca aggtgaagtt catcggcgtg aacttcccct ccgacggccc
     1021 cgtgatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga
     1081 cggcgtgctg aagggcgaga cccacaaggc cctgaagctg aaggacggcg gccactacct
     1141 ggtggagttc aagtccatct acatggccaa gaagcccgtg cagctgcccg gctactacta
     1201 cgtggacgcc aagctggaca tcacctccca caacgaggac tacaccatcg tggagcagta
     1261 cgagcgcacc gagggccgcc accacctgtt cctgagatct ggaaggatcc ccatgcagcc
     1321 agatcccagg cctagcgggg ctggggcctg ctgccgattc ctgcccctgc agtcacagtg
     1381 ccctgagggg gcaggggacg cggtgatgta cgcctccact gagtgcaagg cggaggtgac
     1441 gccctcccag catggcaacc gcaccttcag ctacaccctg gaggatcata ccaagcaggc
     1501 ctttggcatc atgaacgagc tgcggctcag ccagcagctg tgtgacgtca cactgcaggt
     1561 caagtaccag gatgcaccgg ccgcccagtt catggcccac aaggtggtgc tggcctcatc
     1621 cagccctgtc ttcaaggcca tgttcaccaa cgggctgcgg gagcagggca tggaggtggt
     1681 gtccattgag ggtatccacc ccaaggtcat ggagcgcctc attgaattcg cctacacggc
     1741 ctccatctcc atgggcgaga agtgtgtcct ccacgtcatg aacggtgctg tcatgtacca
     1801 gatcgacagc gttgtccgtg cctgcagtga cttcctggtg cagcagctgg accccagcaa
     1861 tgccatcggc atcgccaact tcgctgagca gattggctgt gtggagttgc accagcgtgc
     1921 ccgggagtac atctacatgc attttgggga ggtggccaag caagaggagt tcttcaacct
     1981 gtcccactgc caactggtga ccctcatcag ccgggacgac ctgaacgtgc gctgcgagtc
     2041 cgaggtcttc cacgcctgca tcaactgggt caagtacgac tgcgaacagc gacggttcta
     2101 cgtccaggcg ctgctgcggg ccgtgcgctg ccactcgttg acgccgaact tcctgcagat
     2161 gcagctgcag aagtgcgaga tcctgcagtc cgactcccgc tgcaaggact acctggtcaa
     2221 gatcttcgag gagctcaccc tgcacaagcc cacgcaggtg atgccctgcc gggcgcccaa
     2281 ggtgggccgc ctgatctaca ccgcgggcgg ctacttccga cagtcgctca gctacctgga
     2341 ggcttacaac cccagtgacg gcacctggct ccggttggcg gacctgcagg tgccgcggag
     2401 cggcctggcc ggctgcgtgg tgggcgggct gttgtacgcc gtgggcggca ggaacaactc
     2461 gcccgacggc aacaccgact ccagcgccct ggactgttac aaccccatga ccaatcagtg
     2521 gtcgccctgc gcccccatga gcgtgccccg taaccgcatc ggggtggggg tcatcgatgg
     2581 ccacatctat gccgtcggcg gctcccacgg ctgcatccac cacaacagtg tggagaggta
     2641 tgagccagag cgggatgagt ggcacttggt ggccccaatg ctgacacgaa ggatcggggt
     2701 gggcgtggct gtcctcaatc gtctccttta tgccgtgggg ggctttgacg ggacaaaccg
     2761 ccttaattca gctgagtgtt actacccaga gaggaacgag tggcgaatga tcacagcaat
     2821 gaacaccatc cgaagcgggg caggcgtctg cgtcctgcac aactgtatct atgctgctgg
     2881 gggctatgat ggtcaggacc agctgaacag cgtggagcgc tacgatgtgg aaacagagac
     2941 gtggactttc gtagccccca tgaagcaccg gcgaagtgcc ctggggatca ctgtccacca
     3001 ggggagaatc tacgtccttg gaggctatga tggtcacacg ttcctggaca gtgtggagtg
     3061 ttacgaccca gatacagaca cctggagcga ggtgacccga atgacatcgg gccggagtgg
     3121 ggtgggcgtg gctgtcacca tggagccctg ccggaagcag attgaccagc agaactgtac
     3181 ctgtctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3241 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3301 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3361 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3421 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3481 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3541 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3601 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3661 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3721 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3781 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3841 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3901 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3961 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     4021 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4081 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4141 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4201 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4261 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4321 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4381 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4441 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4501 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4561 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4621 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4681 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4741 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4801 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4861 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4921 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4981 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5041 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5101 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5161 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5221 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5281 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5341 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5401 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5461 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5521 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5581 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5641 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6421 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6481 gattctgtgg ataaccgtat taccgc
//
