LOCUS       sequence-268760-        5203 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 5203)
  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 5203)
  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..5203
                     /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    2233..2367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2257..2276)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2311..2330
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2480..2500)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2480..2498)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2519..2535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2519..2535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2532..2554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2543..2559
                     /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(2567..2573,2574..2591,2592..2597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2612..2633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2750..2767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2921..3509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3001..3020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3680..4471,4472..4540))
                     /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     4303..4322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4541..4645)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4671..5126)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4808..4829)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5020..5039
                     /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 gtggccccag
     1981 tcaaaactgg aaccagggat atagtaacta ttggaatcaa ggctatggca actatggata
     2041 taacagccaa ggttacggtg gttatggagg atatgactac actggttaca acaactacta
     2101 tggatatggt gattatagca accagcagag tggttatggg aaggtatcca ggcgaggtgg
     2161 tcatcaaaat agctacaaac catactaaaa ggcctctcga gcctctagaa ctatagtgag
     2221 tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca aaccacaact
     2281 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2341 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2401 gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc
     2461 cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     2521 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2581 ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2641 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2701 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2761 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2821 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2881 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2941 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3001 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3061 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3121 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3181 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3241 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3301 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3361 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3421 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3601 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3661 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3721 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3781 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3841 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3901 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3961 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4021 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4081 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4141 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4201 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4261 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4321 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4381 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4441 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4501 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4561 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4621 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     4681 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4741 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4801 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4861 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4921 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4981 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5041 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5101 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     5161 agggcgatcg gtgcgggcct cttcgctatt acgccagtcg acc
//
