LOCUS       sequence-262607-        5207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AUF1p42 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 5207)
  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 5207)
  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(p42)
FEATURES             Location/Qualifiers
     source          1..5207
                     /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    1256..1390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1280..1299)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1334..1353
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1503..1523)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1503..1521)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1542..1558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1542..1558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1555..1577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1566..1582
                     /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(1590..1596,1597..1614,1615..1620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1635..1656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1773..1790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1944..2532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2024..2043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2703..3494,3495..3563))
                     /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     3326..3345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3564..3668)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3694..4149)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3831..3852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4043..4062
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4223..5207
                     /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 agggaaaatg tttataggag gccttagctg ggacactaca aagaaagatc
      481 tgaaggacta cttttccaaa tttggtgaag ttgtagactg cactctgaag ttagatccta
      541 tcacagggcg atcaaggggt tttggctttg tgctatttaa agaatcggag agtgtagata
      601 aggtcatgga tcaaaaagaa cataaattga atgggaaggt gattgatcct aaaagggcca
      661 aagccatgaa aacaaaagag ccggttaaaa aaatttttgt tggtggcctt tctccagata
      721 cacctgaaga gaaaataagg gagtactttg gtggttttgg tgaggtggaa tccatagagc
      781 tccccatgga caacaagacc aataagaggc gtgggttctg ctttattacc tttaaggaag
      841 aagaaccagt gaagaagata atggaaaaga aataccacaa tgttggtctt agtaaatgtg
      901 aaataaaagt agccatgtcg aaggaacaat atcagcaaca gcaacagtgg ggatctagag
      961 gaggatttgc aggaagagct cgtggaagag gtggtggccc cagtcaaaac tggaaccagg
     1021 gatatagtaa ctattggaat caaggctatg gcaactatgg atataacagc caaggttacg
     1081 gtggttatgg aggatatgac tacactggtt acaacaacta ctatggatat ggtgattata
     1141 gcaaccagca gagtggttat gggaaggtat ccaggcgagg tggtcatcaa aatagctaca
     1201 aaccatacta aaaggcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     1261 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1321 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1381 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1441 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     1501 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     1561 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1621 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1681 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1741 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1801 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1861 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1921 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1981 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2041 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2101 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2161 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2221 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2281 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2341 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     2401 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2461 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2521 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2581 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2641 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2701 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2761 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2821 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2881 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2941 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3001 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3061 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3121 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3181 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3241 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3301 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3361 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3421 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3481 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3541 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3601 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3661 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     3721 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3781 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3841 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3901 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3961 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4021 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4081 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4141 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4201 cctcttcgct attacgccag tcgaccatag ccaattcaat atggcgtata tggactcatg
     4261 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg
     4321 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg
     4381 ccaattcaat atggcggact tggcactgtg ccaactgggg aggggtctac ttggcacggt
     4441 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt
     4501 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata
     4561 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat
     4621 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc
     4681 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat
     4741 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg
     4801 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg
     4861 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt
     4921 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc
     4981 tattaatagt aacttggcaa gtacattact attggaagga cgccagggta cattggcagt
     5041 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt
     5101 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta
     5161 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
