LOCUS       sequence-268769-        5260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AUF1p45 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 5260)
  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 5260)
  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(p45)
FEATURES             Location/Qualifiers
     source          1..5260
                     /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    2290..2424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2314..2333)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2368..2387
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2537..2557)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2537..2555)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2576..2592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2576..2592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2589..2611)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2600..2616
                     /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(2624..2630,2631..2648,2649..2654))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2669..2690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2807..2824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2978..3566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3058..3077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3737..4528,4529..4597))
                     /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     4360..4379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4598..4702)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4728..5183)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4865..4886)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5077..5096
                     /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 ccattcaaac tcctccccac gacactctga
     1441 agcagcgacg gcacagcggg aagaatggaa aatgtttata ggaggcctta gctgggacac
     1501 tacaaagaaa gatctgaagg actacttttc caaatttggt gaagttgtag actgcactct
     1561 gaagttagat cctatcacag ggcgatcaag gggttttggc tttgtgctat ttaaagaatc
     1621 ggagagtgta gataaggtca tggatcaaaa agaacataaa ttgaatggga aggtgattga
     1681 tcctaaaagg gccaaagcca tgaaaacaaa agagccggtt aaaaaaattt ttgttggtgg
     1741 cctttctcca gatacacctg aagagaaaat aagggagtac tttggtggtt ttggtgaggt
     1801 ggaatccata gagctcccca tggacaacaa gaccaataag aggcgtgggt tctgctttat
     1861 tacctttaag gaagaagaac cagtgaagaa gataatggaa aagaaatacc acaatgttgg
     1921 tcttagtaaa tgtgaaataa aagtagccat gtcgaaggaa caatatcagc aacagcaaca
     1981 gtggggatct agaggaggat ttgcaggaag agctcgtgga agaggtggtg gccccagtca
     2041 aaactggaac cagggatata gtaactattg gaatcaaggc tatggcaact atggatataa
     2101 cagccaaggt tacggtggtt atggaggata tgactacact ggttacaaca actactatgg
     2161 atatggtgat tatagcaacc agcagagtgg ttatgggaag gtatccaggc gaggtggtca
     2221 tcaaaatagc tacaaaccat actaaaaggc ctctcgagcc tctagaacta tagtgagtcg
     2281 tattacgtag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2341 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2401 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2461 cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg
     2521 tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     2581 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga
     2641 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2701 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3661 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3721 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3781 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3841 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3901 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3961 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4021 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4081 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4141 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4201 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4261 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4321 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4381 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4441 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4501 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4561 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4681 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     4741 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4801 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4861 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     4921 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     4981 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5041 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5101 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5161 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
     5221 gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
//
