LOCUS       sequence-236007-        6834 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 6834)
  AUTHORS   Camp ND, James RG, Dawson DW, Yan F, Davison JM, Houck SA, Tang X, 
            Zheng N, Major MB, Moon RT
  TITLE     Wilms tumor gene on X chromosome (WTX) inhibits degradation of NRF2 
            protein through competitive binding to KEAP1 protein.
  JOURNAL   J Biol Chem. 2012 Feb 24;287(9):6539-50. Epub 2012 Jan 3.
  PUBMED    22215675
REFERENCE   2  (bases 1 to 6834)
  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: "J Biol 
            Chem."; date: "2012-02-24"; volume: "287(9)"; pages: "6539-50. Epub 
            2012 Jan 3"
COMMENT     Sequence Label: NC16 pCDNA3.1 FLAG NRF2
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      701..710
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             710..733
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2608..2625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2614..2838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2884..3312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2971..2990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3181..3202
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3321..3341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3326..3655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3506..3641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3568..3587
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3703..3750
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3769..4143
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4273..4394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4310..4329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4364..4383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4443..4459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4443..4459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4456..4478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4467..4483
                     /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(4491..4497,4498..4515,4516..4521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4536..4557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4674..4691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4845..5433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4925..4944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5604..6395,6396..6464))
                     /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     6227..6246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6465..6569)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6644..6663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc gccaccatgg actacaagga
      721 cgacgacgac aagggcggcg cgcctatgat ggacttggag ctgccgccgc cgggactccc
      781 gtcccagcag gacatggatt tgattgacat actttggagg caagatatag atcttggagt
      841 aagtcgagaa gtatttgact tcagtcagcg acggaaagag tatgagctgg aaaaacagaa
      901 aaaacttgaa aaggaaagac aagaacaact ccaaaaggag caagagaaag cctttttcgc
      961 tcagttacaa ctagatgaag agacaggtga atttctccca attcagccag cccagcacat
     1021 ccagtcagaa accagtggat ctgccaacta ctcccaggtt gcccacattc ccaaatcaga
     1081 tgctttgtac tttgatgact gcatgcagct tttggcgcag acattcccgt ttgtagatga
     1141 caatgaggtt tcttcggcta cgtttcagtc acttgttcct gatattcccg gtcacatcga
     1201 gagcccagtc ttcattgcta ctaatcaggc tcagtcacct gaaacttctg ttgctcaggt
     1261 agcccctgtt gatttagacg gtatgcaaca ggacattgag caagtttggg aggagctatt
     1321 atccattcct gagttacagt gtcttaatat tgaaaatgac aagctggttg agactaccat
     1381 ggttccaagt ccagaagcca aactgacaga agttgacaat tatcattttt actcatctat
     1441 accctcaatg gaaaaagaag taggtaactg tagtccacat tttcttaatg cttttgagga
     1501 ttccttcagc agcatcctct ccacagaaga ccccaaccag ttgacagtga actcattaaa
     1561 ttcagatgcc acagtcaaca cagattttgg tgatgaattt tattctgctt tcatagctga
     1621 gcccagtatc agcaacagca tgccctcacc tgctacttta agccattcac tctctgaact
     1681 tctaaatggg cccattgatg tttctgatct atcactttgc aaagctttca accaaaacca
     1741 ccctgaaagc acagcagaat tcaatgattc tgactccggc atttcactaa acacaagtcc
     1801 cagtgtggca tcaccagaac actcagtgga atcttccagc tatggagaca cactacttgg
     1861 cctcagtgat tctgaagtgg aagagctaga tagtgcccct ggaagtgtca aacagaatgg
     1921 tcctaaaaca ccagtacatt cttctgggga tatggtacaa cccttgtcac catctcaggg
     1981 gcagagcact cacgtgcatg atgcccaatg tgagaacaca ccagagaaag aattgcctgt
     2041 aagtcctggt catcggaaaa ccccattcac aaaagacaaa cattcaagcc gcttggaggc
     2101 tcatctcaca agagatgaac ttagggcaaa agctctccat atcccattcc ctgtagaaaa
     2161 aatcattaac ctccctgttg ttgacttcaa cgaaatgatg tccaaagagc agttcaatga
     2221 agctcaactt gcattaattc gggatatacg taggaggggt aagaataaag tggctgctca
     2281 gaattgcaga aaaagaaaac tggaaaatat agtagaacta gagcaagatt tagatcattt
     2341 gaaagatgaa aaagaaaaat tgctcaaaga aaaaggagaa aatgacaaaa gccttcacct
     2401 actgaaaaaa caactcagca ccttatatct cgaagttttc agcatgctac gtgatgaaga
     2461 tggaaaacct tattctccta gtgaatactc cctgcagcaa acaagagatg gcaatgtttt
     2521 ccttgttccc aaaagtaaga agccagatgt taagaaaaac taggcggccg ctcgagtcta
     2581 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2641 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2701 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2761 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2821 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2881 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2941 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3001 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3061 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3121 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3181 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3241 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3301 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3361 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3421 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3481 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3541 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3601 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3661 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     3721 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     3781 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     3841 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     3901 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     3961 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     4021 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     4081 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     4141 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4201 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4261 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4321 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4381 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4441 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4501 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4561 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4621 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4681 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4741 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4801 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4861 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4921 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4981 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5041 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5101 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5161 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5221 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5281 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5461 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5521 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5581 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5641 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5701 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5761 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5821 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5881 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5941 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6001 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6061 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6121 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6181 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6241 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6301 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6361 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6421 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6481 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6541 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6601 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6661 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6721 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     6781 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
