LOCUS       sequence-262559-        5184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HuR 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 5184)
  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 5184)
  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 Flag3HuR
FEATURES             Location/Qualifiers
     source          1..5184
                     /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             134..199
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1233..1367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1257..1276)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1311..1330
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1480..1500)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1480..1498)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1519..1535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1519..1535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1532..1554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1543..1559
                     /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(1567..1573,1574..1591,1592..1597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1612..1633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1750..1767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1921..2509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2001..2020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2680..3471,3472..3540))
                     /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     3303..3322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3541..3645)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3671..4126)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3808..3829)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4020..4039
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4200..5184
                     /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 atggattata aagatcatga tggcgattat aaagatcatg atattgatta
      181 taaagatgat gatgataaaa tgtctaatgg ttatgaagac cacatggccg aagactgcag
      241 gggtgacatc gggagaacga atttgatcgt caactacctc cctcagaaca tgacccagga
      301 tgagttacga agcctgttca gcagcattgg tgaagttgaa tctgcaaaac ttattcggga
      361 taaagtagca ggacacagct tgggctatgg ctttgtgaac tacgtgaccg cgaaggatgc
      421 agagagagcg atcaacacgc tgaacggctt gaggctccag tcaaaaacca ttaaggtgtc
      481 gtatgctcgc ccgagctcag aggtgatcaa agacgccaac ttgtacatca gcgggctccc
      541 gcggaccatg acccagaagg acgtagaaga catgttctct cggtttgggc ggatcatcaa
      601 ctcgcgggtc ctcgtggatc agactacagg tttgtccaga ggggttgcgt ttatccggtt
      661 tgacaaacgg tcggaggcag aagaggcaat taccagtttc aatggtcata aacccccagg
      721 ttcctctgag cccatcacag tgaagtttgc agccaacccc aaccagaaca aaaacgtggc
      781 actcctctcg cagctgtacc actcgccagc gcgacggttc ggaggccccg ttcaccacca
      841 ggcgcagaga ttcaggttct cccccatggg cgtcgatcac atgagcgggc tctctggcgt
      901 caacgtgcca ggaaacgcct cctccggctg gtgcattttc atctacaacc tggggcagga
      961 tgccgacgag gggatcctct ggcagatgtt tgggccgttt ggtgccgtca ccaatgtgaa
     1021 agtgatccgc gacttcaaca ccaacaagtg caaagggttt ggctttgtga ccatgacaaa
     1081 ctatgaagaa gccgcgatgg ccatagccag cctgaacggc taccgcctgg gggacaaaat
     1141 cttacaggtt tccttcaaaa ccaacaagtc ccacaaataa tcgaattcaa ggcctctcga
     1201 gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
     1261 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1321 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1381 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
     1441 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     1501 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1561 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1621 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1681 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1741 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1801 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1861 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1921 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1981 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2041 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2101 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2161 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2221 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2281 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2341 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2401 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2461 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2521 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2581 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2641 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2701 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2761 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2821 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2881 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2941 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3001 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3061 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3121 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3181 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3241 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3301 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3361 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3421 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3481 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3541 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3601 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3661 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3721 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3781 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3841 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3901 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3961 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4021 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4081 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     4141 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
     4201 accatagcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
     4261 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
     4321 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacttgg
     4381 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
     4441 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
     4501 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
     4561 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
     4621 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
     4681 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
     4741 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
     4801 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
     4861 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
     4921 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
     4981 cattactatt ggaaggacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
     5041 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
     5101 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
     5161 ataagcaatg ctcgtttagg gaac
//
