LOCUS       sequence-262606-        5117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AUF1p40 in mammalian cells. Can also be used to generate 
            mRNA for zebrafish expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5117)
  AUTHORS   Poganik JR, Long MJC, Disare MT, Liu X, Chang SH, Hla T, Aye Y
  TITLE     Post-transcriptional regulation of Nrf2-mRNA by the mRNA-binding 
            proteins HuR and AUF1.
  JOURNAL   FASEB J. 2019 Nov 1:fj201901930R. doi: 10.1096/fj.201901930R.
  PUBMED    31665914
REFERENCE   2  (bases 1 to 5117)
  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: "FASEB J. 
            2019 Nov 1:fj201901930R. doi: 10.1096/fj.201901930R."
COMMENT     Sequence Label: pCS2+8 Flag2AUF1(p40)
FEATURES             Location/Qualifiers
     source          1..5117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             147..194
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     regulatory      286..295
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1166..1300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1190..1209)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1244..1263
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1413..1433)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1413..1431)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1452..1468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1452..1468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1465..1487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1476..1492
                     /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(1500..1506,1507..1524,1525..1530))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1545..1566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1683..1700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1854..2442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1934..1953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2613..3404,3405..3473))
                     /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     3236..3255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3474..3578)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3604..4059)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3741..3762)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3953..3972
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4133..5117
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccactagt ggcgcgccat taattaaagg ccggccagcg
      121 atcgccggac atcgattccc accatggact ataaggatga cgacgataag gattacaaag
      181 acgatgatga caaagggtcc atgtcggagg agcagttcgg cggggacggg gcggcggcag
      241 cggcaacggc ggcggtaggc ggctcggcgg gcgagcagga gggagccatg gtggcggcga
      301 cacagggggc agcggcggcg gcgggaagcg gagccgggac cgggggcgga accgcgtctg
      361 gaggcaccga agggggcagc gccgagtcgg agggggcgaa gattgacgcc agtaagaacg
      421 aggaggatga aggccattca aactcctccc cacgacactc tgaagcagcg acggcacagc
      481 gggaagaatg gaaaatgttt ataggaggcc ttagctggga cactacaaag aaagatctga
      541 aggactactt ttccaaattt ggtgaagttg tagactgcac tctgaagtta gatcctatca
      601 cagggcgatc aaggggtttt ggctttgtgc tatttaaaga atcggagagt gtagataagg
      661 tcatggatca aaaagaacat aaattgaatg ggaaggtgat tgatcctaaa agggccaaag
      721 ccatgaaaac aaaagagccg gttaaaaaaa tttttgttgg tggcctttct ccagatacac
      781 ctgaagagaa aataagggag tactttggtg gttttggtga ggtggaatcc atagagctcc
      841 ccatggacaa caagaccaat aagaggcgtg ggttctgctt tattaccttt aaggaagaag
      901 aaccagtgaa gaagataatg gaaaagaaat accacaatgt tggtcttagt aaatgtgaaa
      961 taaaagtagc catgtcgaag gaacaatatc agcaacagca acagtgggga tctagaggag
     1021 gatttgcagg aagagctcgt ggaagaggtg gtgaccagca gagtggttat gggaaggtat
     1081 ccaggcgagg tggtcatcaa aatagctaca aaccatacta aaaggcctct cgagcctcta
     1141 gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg
     1201 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1261 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1321 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc
     1381 aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     1441 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1501 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1561 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1621 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2281 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2641 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2701 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2761 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2821 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2881 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2941 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3001 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3061 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3121 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3181 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3241 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3301 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3361 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3421 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3481 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3541 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3601 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3661 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3721 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3781 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3841 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3901 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3961 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4021 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     4081 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccatag
     4141 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     4201 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     4261 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggact tggcactgtg
     4321 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     4381 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     4441 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     4501 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     4561 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     4621 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     4681 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     4741 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     4801 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     4861 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     4921 attggaagga cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
     4981 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
     5041 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
     5101 atgctcgttt agggaac
//
