LOCUS       sequence-232076-        6104 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 6104)
  AUTHORS   Fan W, Tang Z, Chen D, Moughon D, Ding X, Chen S, Zhu M, Zhong Q
  TITLE     Keap1 facilitates p62-mediated ubiquitin aggregate clearance via 
            autophagy.
  JOURNAL   Autophagy. 2010 Jul 22. 6(5):.
  PUBMED    20495340
REFERENCE   2  (bases 1 to 6104)
  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: "Autophagy. 
            2010 Jul 22. 6(5):."
COMMENT     Sequence Label: hrGFP-Keap1
FEATURES             Location/Qualifiers
     source          1..6104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             654..1370
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3329..3348)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3330..3348)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3622..3743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3659..3678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3713..3732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3750..4205)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3887..3908)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4099..4118
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4232..4336
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    4351..4384
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(4426..5217,5218..5286))
                     /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     5049..5068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5456..6044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5945..5964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc accatggtga
      661 gcaagcagat cctgaagaac accggcctgc aggagatcat gagcttcaag gtgaacctgg
      721 agggcgtggt gaacaaccac gtgttcacca tggagggctg cggcaagggc aacatcctgt
      781 tcggcaacca gctggtgcag atccgcgtga ccaagggcgc ccccctgccc ttcgccttcg
      841 acatcctgag ccccgccttc cagtacggca accgcacctt caccaagtac cccgaggaca
      901 tcagcgactt cttcatccag agcttccccg ccggcttcgt gtacgagcgc accctgcgct
      961 acgaggacgg cggcctggtg gagatccgca gcgacatcaa cctgatcgag gagatgttcg
     1021 tgtaccgcgt ggagtacaag ggccgcaact tccccaacga cggccccgtg atgaagaaga
     1081 ccatcaccgg cctgcagccc agcttcgagg tggtgtacat gaacgacggc gtgctggtgg
     1141 gccaggtgat cctggtgtac cgcctgaaca gcggcaagtt ctacagctgc cacatgcgca
     1201 ccctgatgaa gagcaagggc gtggtgaagg acttccccga gtaccacttc atccagcacc
     1261 gcctggagaa gacctacgtg gaggacggcg gcttcgtgga gcagcacgag accgccatcg
     1321 cccagctgac cagcctgggc aagcccctgg gcagcctgca cgagtgggtg gagctctccg
     1381 gactcagatc tcgagctgaa gcttcgcagc cagatcccag gcctagcggg gctggggcct
     1441 gctgccgatt cctgcccctg cagtcacagt gccctgaggg ggcaggggac gcggtgatgt
     1501 acgcctccac tgagtgcaag gcggaggtga cgccctccca gcatggcaac cgcaccttca
     1561 gctacaccct ggaggatcat accaagcagg cctttggcat catgaacgag ctgcggctca
     1621 gccagcagct gtgtgacgtc acactgcagg tcaagtacca ggatgcaccg gccgcccagt
     1681 tcatggccca caaggtggtg ctggcctcat ccagccctgt cttcaaggcc atgttcacca
     1741 acgggctgcg ggagcagggc atggaggtgg tgtccattga gggtatccac cccaaggtca
     1801 tggagcgcct cattgaattc gcctacacgg cctccatctc catgggcgag aagtgtgtcc
     1861 tccacgtcat gaacggtgct gtcatgtacc agatcgacag cgttgtccgt gcctgcagtg
     1921 acttcctggt gcagcagctg gaccccagca atgccatcgg catcgccaac ttcgctgagc
     1981 agattggctg tgtggagttg caccagcgtg cccgggagta catctacatg cattttgggg
     2041 aggtggccaa gcaagaggag ttcttcaacc tgtcccactg ccaactggtg accctcatca
     2101 gccgggacga cctgaacgtg cgctgcgagt ccgaggtctt ccacgcctgc atcaactggg
     2161 tcaagtacga ctgcgaacag cgacggttct acgtccaggc gctgctgcgg gccgtgcgct
     2221 gccactcgtt gacgccgaac ttcctgcaga tgcagctgca gaagtgcgag atcctgcagt
     2281 ccgactcccg ctgcaaggac tacctggtca agatcttcga ggagctcacc ctgcacaagc
     2341 ccacgcaggt gatgccctgc cgggcgccca aggtgggccg cctgatctac accgcgggcg
     2401 gctacttccg acagtcgctc agctacctgg aggcttacaa ccccagtgac ggcacctggc
     2461 tccggttggc ggacctgcag gtgccgcgga gcggcctggc cggctgcgtg gtgggcgggc
     2521 tgttgtacgc cgtgggcggc aggaacaact cgcccgacgg caacaccgac tccagcgccc
     2581 tggactgtta caaccccatg accaatcagt ggtcgccctg cgcccccatg agcgtgcccc
     2641 gtaaccgcat cggggtgggg gtcatcgatg gccacatcta tgccgtcggc ggctcccacg
     2701 gctgcatcca ccacaacagt gtggagaggt atgagccaga gcgggatgag tggcacttgg
     2761 tggccccaat gctgacacga aggatcgggg tgggcgtggc tgtcctcaat cgtctccttt
     2821 atgccgtggg gggctttgac gggacaaacc gccttaattc agctgagtgt tactacccag
     2881 agaggaacga gtggcgaatg atcacagcaa tgaacaccat ccgaagcggg gcaggcgtct
     2941 gcgtcctgca caactgtatc tatgctgctg ggggctatga tggtcaggac cagctgaaca
     3001 gcgtggagcg ctacgatgtg gaaacagaga cgtggacttt cgtagccccc atgaagcacc
     3061 ggcgaagtgc cctggggatc actgtccacc aggggagaat ctacgtcctt ggaggctatg
     3121 atggtcacac gttcctggac agtgtggagt gttacgaccc agatacagac acctggagcg
     3181 aggtgacccg aatgacatcg ggccggagtg gggtgggcgt ggctgtcacc atggagccct
     3241 gccggaagca gattgaccag cagaactgta cctgttgagg atccaccgga tctagataat
     3301 agggtaccag gtaagtgtac ccaattcgcc ctatagtgag tcgtattaca attcactcga
     3361 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg agatccaatt tttaagtgta
     3421 taatgtgtta aactactgat tctaattgtt tgtgtatttt agattcacag tcccaaggct
     3481 catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca taccacattt
     3541 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3601 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3661 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3721 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     3781 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3841 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3901 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3961 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4021 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4081 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4141 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4201 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4261 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4321 ttgaaaaagg aagaatcctg aggccgggcc ataacttcgt ataatgtatg ctatacgaag
     4381 ttatccatgg gccccccctc gacatgagta aacttggtct gacagttacc aatgcttaat
     4441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4741 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5041 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5281 actcatactc ttcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5341 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5401 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5461 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5521 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5581 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5641 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5701 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5761 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5821 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5881 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5941 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6001 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6061 cctttttacg gttcctggcc ttttgctggc cttttgctca catg
//
