LOCUS       sequence-268761-        5056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AUF1p37 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 5056)
  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 5056)
  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(p37)
FEATURES             Location/Qualifiers
     source          1..5056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1124..1171
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     regulatory      1263..1272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2086..2220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2110..2129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2164..2183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2333..2353)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2333..2351)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2372..2388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2372..2388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2385..2407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2396..2412
                     /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(2420..2426,2427..2444,2445..2450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2465..2486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2603..2620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2774..3362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2854..2873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3533..4324,4325..4393))
                     /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     4156..4175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4394..4498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4524..4979)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4661..4682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4873..4892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccactagtgg ccgccattaa
     1081 ttaaaggccg gccagcgatc gccggacatc gattcccacc atggactata aggatgacga
     1141 cgataaggat tacaaagacg atgatgacaa agggtccatg tcggaggagc agttcggcgg
     1201 ggacggggcg gcggcagcgg caacggcggc ggtaggcggc tcggcgggcg agcaggaggg
     1261 agccatggtg gcggcgacac agggggcagc ggcggcggcg ggaagcggag ccgggaccgg
     1321 gggcggaacc gcgtctggag gcaccgaagg gggcagcgcc gagtcggagg gggcgaagat
     1381 tgacgccagt aagaacgagg aggatgaagg gaaaatgttt ataggaggcc ttagctggga
     1441 cactacaaag aaagatctga aggactactt ttccaaattt ggtgaagttg tagactgcac
     1501 tctgaagtta gatcctatca cagggcgatc aaggggtttt ggctttgtgc tatttaaaga
     1561 atcggagagt gtagataagg tcatggatca aaaagaacat aaattgaatg ggaaggtgat
     1621 tgatcctaaa agggccaaag ccatgaaaac aaaagagccg gttaaaaaaa tttttgttgg
     1681 tggcctttct ccagatacac ctgaagagaa aataagggag tactttggtg gttttggtga
     1741 ggtggaatcc atagagctcc ccatggacaa caagaccaat aagaggcgtg ggttctgctt
     1801 tattaccttt aaggaagaag aaccagtgaa gaagataatg gaaaagaaat accacaatgt
     1861 tggtcttagt aaatgtgaaa taaaagtagc catgtcgaag gaacaatatc agcaacagca
     1921 acagtgggga tctagaggag gatttgcagg aagagctcgt ggaagaggtg gtgaccagca
     1981 gagtggttat gggaaggtat ccaggcgagg tggtcatcaa aatagctaca aaccatacta
     2041 aaaggcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc agacatgata
     2101 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2161 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2221 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2281 taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt
     2341 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2401 atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg
     2461 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2521 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4501 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     4561 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4621 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4681 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4741 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4801 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4861 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4921 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4981 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5041 attacgccag tcgacc
//
