LOCUS       sequence-268767-        5113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AUF1p40 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 5113)
  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 5113)
  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(p40)
FEATURES             Location/Qualifiers
     source          1..5113
                     /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    2143..2277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2167..2186)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2221..2240
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2390..2410)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2390..2408)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2429..2445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2429..2445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2442..2464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2453..2469
                     /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(2477..2483,2484..2501,2502..2507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2522..2543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2660..2677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2831..3419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2911..2930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3590..4381,4382..4450))
                     /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     4213..4232
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4451..4555)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4581..5036)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4718..4739)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4930..4949
                     /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 accagcagag
     2041 tggttatggg aaggtatcca ggcgaggtgg tcatcaaaat agctacaaac catactaaaa
     2101 ggcctctcga gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
     2161 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2221 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2281 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2341 ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     2401 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2461 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     2521 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2581 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2641 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2701 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2761 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2821 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2881 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2941 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3001 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3061 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3121 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3181 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3241 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3301 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3361 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3421 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3481 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3541 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3601 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3661 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3721 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3781 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3841 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3901 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3961 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4021 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4081 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4141 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4201 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4261 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4321 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4381 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4441 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4501 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4561 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4621 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4681 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4741 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4801 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4861 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4921 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4981 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     5041 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5101 acgccagtcg acc
//
