LOCUS       sequence-262610-        5264 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 5264)
  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 5264)
  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..5264
                     /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             147..194
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     regulatory      286..295
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1313..1447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1337..1356)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1391..1410
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1560..1580)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1560..1578)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1599..1615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1599..1615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1612..1634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1623..1639
                     /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(1647..1653,1654..1671,1672..1677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1692..1713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1830..1847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2001..2589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2081..2100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2760..3551,3552..3620))
                     /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     3383..3402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3621..3725)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3751..4206)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3888..3909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4100..4119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4280..5264
                     /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 atcgattccc accatggact ataaggatga cgacgataag gattacaaag
      181 acgatgatga caaagggtcc atgtcggagg agcagttcgg cggggacggg gcggcggcag
      241 cggcaacggc ggcggtaggc ggctcggcgg gcgagcagga gggagccatg gtggcggcga
      301 cacagggggc agcggcggcg gcgggaagcg gagccgggac cgggggcgga accgcgtctg
      361 gaggcaccga agggggcagc gccgagtcgg agggggcgaa gattgacgcc agtaagaacg
      421 aggaggatga aggccattca aactcctccc cacgacactc tgaagcagcg acggcacagc
      481 gggaagaatg gaaaatgttt ataggaggcc ttagctggga cactacaaag aaagatctga
      541 aggactactt ttccaaattt ggtgaagttg tagactgcac tctgaagtta gatcctatca
      601 cagggcgatc aaggggtttt ggctttgtgc tatttaaaga atcggagagt gtagataagg
      661 tcatggatca aaaagaacat aaattgaatg ggaaggtgat tgatcctaaa agggccaaag
      721 ccatgaaaac aaaagagccg gttaaaaaaa tttttgttgg tggcctttct ccagatacac
      781 ctgaagagaa aataagggag tactttggtg gttttggtga ggtggaatcc atagagctcc
      841 ccatggacaa caagaccaat aagaggcgtg ggttctgctt tattaccttt aaggaagaag
      901 aaccagtgaa gaagataatg gaaaagaaat accacaatgt tggtcttagt aaatgtgaaa
      961 taaaagtagc catgtcgaag gaacaatatc agcaacagca acagtgggga tctagaggag
     1021 gatttgcagg aagagctcgt ggaagaggtg gtggccccag tcaaaactgg aaccagggat
     1081 atagtaacta ttggaatcaa ggctatggca actatggata taacagccaa ggttacggtg
     1141 gttatggagg atatgactac actggttaca acaactacta tggatatggt gattatagca
     1201 accagcagag tggttatggg aaggtatcca ggcgaggtgg tcatcaaaat agctacaaac
     1261 catactaaaa ggcctctcga gcctctagaa ctatagtgag tcgtattacg tagatccaga
     1321 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1381 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1441 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1501 ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc
     1561 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     1621 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     1681 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     1741 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2161 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2401 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     2461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2701 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2761 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2821 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2881 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2941 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3001 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3061 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3121 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3181 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3241 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3301 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3361 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3421 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3481 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3541 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3601 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3661 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3721 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     3781 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3841 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3901 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3961 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4021 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4081 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4141 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4201 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4261 cttcgctatt acgccagtcg accatagcca attcaatatg gcgtatatgg actcatgcca
     4321 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     4381 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     4441 attcaatatg gcggacttgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     4501 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     4561 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     4621 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     4681 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     4741 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     4801 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     4861 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     4921 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     4981 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     5041 taatagtaac ttggcaagta cattactatt ggaaggacgc cagggtacat tggcagtact
     5101 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     5161 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     5221 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaac
//
