LOCUS       sequence-268770-        5181 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 5181)
  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 5181)
  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..5181
                     /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             1112..1177
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2211..2345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2235..2254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2289..2308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2458..2478)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2458..2476)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2497..2513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2497..2513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2510..2532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2521..2537
                     /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(2545..2551,2552..2569,2570..2575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2590..2611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2728..2745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2899..3487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2979..2998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3658..4449,4450..4518))
                     /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     4281..4300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4519..4623)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4649..5104)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4786..4807)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4998..5017
                     /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 cgcgccatta
     1081 attaaaggcc ggccagcgat cgccggacat ggattataaa gatcatgatg gcgattataa
     1141 agatcatgat attgattata aagatgatga tgataaaatg tctaatggtt atgaagacca
     1201 catggccgaa gactgcaggg gtgacatcgg gagaacgaat ttgatcgtca actacctccc
     1261 tcagaacatg acccaggatg agttacgaag cctgttcagc agcattggtg aagttgaatc
     1321 tgcaaaactt attcgggata aagtagcagg acacagcttg ggctatggct ttgtgaacta
     1381 cgtgaccgcg aaggatgcag agagagcgat caacacgctg aacggcttga ggctccagtc
     1441 aaaaaccatt aaggtgtcgt atgctcgccc gagctcagag gtgatcaaag acgccaactt
     1501 gtacatcagc gggctcccgc ggaccatgac ccagaaggac gtagaagaca tgttctctcg
     1561 gtttgggcgg atcatcaact cgcgggtcct cgtggatcag actacaggtt tgtccagagg
     1621 ggttgcgttt atccggtttg acaaacggtc ggaggcagaa gaggcaatta ccagtttcaa
     1681 tggtcataaa cccccaggtt cctctgagcc catcacagtg aagtttgcag ccaaccccaa
     1741 ccagaacaaa aacgtggcac tcctctcgca gctgtaccac tcgccagcgc gacggttcgg
     1801 aggccccgtt caccaccagg cgcagagatt caggttctcc cccatgggcg tcgatcacat
     1861 gagcgggctc tctggcgtca acgtgccagg aaacgcctcc tccggctggt gcattttcat
     1921 ctacaacctg gggcaggatg ccgacgaggg gatcctctgg cagatgtttg ggccgtttgg
     1981 tgccgtcacc aatgtgaaag tgatccgcga cttcaacacc aacaagtgca aagggtttgg
     2041 ctttgtgacc atgacaaact atgaagaagc cgcgatggcc atagccagcc tgaacggcta
     2101 ccgcctgggg gacaaaatct tacaggtttc cttcaaaacc aacaagtccc acaaataatc
     2161 gaattcaagg cctctcgagc ctctagaact atagtgagtc gtattacgta gatccagaca
     2221 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2281 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2341 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2401 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     2461 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2521 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     2581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4621 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4681 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4741 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4801 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4861 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4921 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4981 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5041 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5101 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5161 tcgctattac gccagtcgac c
//
