LOCUS       sequence-262560-        5060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AUF1p37 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 5060)
  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 5060)
  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(p37)
FEATURES             Location/Qualifiers
     source          1..5060
                     /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    1109..1243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1133..1152)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1187..1206
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1356..1376)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1356..1374)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1395..1411)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1395..1411)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1408..1430)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1419..1435
                     /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(1443..1449,1450..1467,1468..1473))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1488..1509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1626..1643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1797..2385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1877..1896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2556..3347,3348..3416))
                     /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     3179..3198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3417..3521)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3547..4002)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3684..3705)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3896..3915
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4076..5060
                     /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 agggaaaatg tttataggag gccttagctg ggacactaca aagaaagatc
      481 tgaaggacta cttttccaaa tttggtgaag ttgtagactg cactctgaag ttagatccta
      541 tcacagggcg atcaaggggt tttggctttg tgctatttaa agaatcggag agtgtagata
      601 aggtcatgga tcaaaaagaa cataaattga atgggaaggt gattgatcct aaaagggcca
      661 aagccatgaa aacaaaagag ccggttaaaa aaatttttgt tggtggcctt tctccagata
      721 cacctgaaga gaaaataagg gagtactttg gtggttttgg tgaggtggaa tccatagagc
      781 tccccatgga caacaagacc aataagaggc gtgggttctg ctttattacc tttaaggaag
      841 aagaaccagt gaagaagata atggaaaaga aataccacaa tgttggtctt agtaaatgtg
      901 aaataaaagt agccatgtcg aaggaacaat atcagcaaca gcaacagtgg ggatctagag
      961 gaggatttgc aggaagagct cgtggaagag gtggtgacca gcagagtggt tatgggaagg
     1021 tatccaggcg aggtggtcat caaaatagct acaaaccata ctaaaaggcc tctcgagcct
     1081 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     1141 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1201 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1261 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     1321 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1381 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1441 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1501 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1561 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1621 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1681 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1741 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1801 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1861 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1921 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1981 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2041 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2101 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2161 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2221 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2281 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2341 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2401 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2461 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2521 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2581 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2641 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2701 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2761 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2821 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2881 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2941 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3001 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3061 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3121 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3181 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3241 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3301 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3361 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3421 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3481 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3541 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3601 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3661 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3721 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3781 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3841 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3901 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3961 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     4021 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacca
     4081 tagccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
     4141 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
     4201 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acttggcact
     4261 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
     4321 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     4381 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     4441 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     4501 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     4561 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     4621 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     4681 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     4741 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     4801 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     4861 actattggaa ggacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     4921 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     4981 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     5041 gcaatgctcg tttagggaac
//
