LOCUS       sequence-194151-        5860 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Reporter for mouse Fgf21 promoter activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5860)
  AUTHORS   Miyake M, Nomura A, Ogura A, Takehana K, Kitahara Y, Takahara K, 
            Tsugawa K, Miyamoto C, Miura N, Sato R, Kurahashi K, Harding HP, 
            Oyadomari M, Ron D, Oyadomari S
  TITLE     Skeletal muscle-specific eukaryotic translation initiation factor 
            2alpha phosphorylation controls amino acid metabolism and fibroblast
            growth factor 21-mediated non-cell-autonomous energy metabolism.
  JOURNAL   FASEB J. 2016 Feb;30(2):798-812. doi: 10.1096/fj.15-275990. Epub 
            2015 Oct 20.
  PUBMED    26487695
REFERENCE   2  (bases 1 to 5860)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1096/fj.15-275990"; journalName: "FASEB J"; date: "2016-02"; 
            volume: "30"; issue: "2"; pages: "798-812"
COMMENT     Sequence Label: pGL3-mFgf21pro2-luc
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1124..1133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1130..2782
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1196..1214)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2823..2944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2834..2853)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2888..2907
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3192..3209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3363..3951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3443..3462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4122..4913,4914..4982))
                     /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     4745..4764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4983..5087)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5114..5569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5201..5220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5411..5432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5700..5748
                     /note="synthetic polyadenylation signal"
     misc_feature    5762..5853
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5802..5821
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagccagc tgggagactt ccacctgctg ggctgaggac
       61 tcctcttaca cactgctgat acagctctcc tgatgaaaga aagcctcaga cacgcccccc
      121 ccccaaagca ccttgaagct taaaattcca aaaggatgaa ccacaccagc tcagttgctt
      181 acacagaaag tctgaagccc caggttcctt cttagtaccc aagagccaga ccctagcctc
      241 ctccctcaga cccaagagct agatcccagc ctcctccctc agacccaaga gccagaccct
      301 agcctcctcc ctcagatcca ggaatacaga ccccagccac ctccctcaga cccagaactg
      361 cagagcccag ccaccttcag acccaggagt gtagacttca gtctcctacc tcagacccag
      421 aactgcagac cccagctacc ttcagaccca ggagtgtaga ccccagcctc ctccttcaga
      481 ctcaggagtg tagaccccag cctcctccct cagactcagg agtgcagacc ctagcctcct
      541 ccctcagact caggagtgca gaccccagcc tcctccttca gactcaggag tgcagacaag
      601 tccccttcct tagacccaag gccccaaacc ccatctttcc tcccctcaga tactaaggtg
      661 aagatcccaa cctcctccag agtccaaccc tcctccctca gaccaaggag cacagacccc
      721 agcctcctcc tccagattta ggagtgcaga cccgcccccc agcactcttc ccccccaaac
      781 ccagggggcc catgcctagc ccttttcatt cagacccctg ttggaaagac ccccccattg
      841 catcatccgt ccaggccgcc ctggccacgg tggaattcag gttcctgcca agtgtgtcaa
      901 atatcacgcg tcaggagtgg ggagggcacg tgggcgggcc tgtctgggta taaattctgg
      961 tatttctgcg ttcaccagac agccttagtg tcttctcagc tggggattca acacaggaga
     1021 aacagccatt cactttgcct gagccccagt ctgaacctga cccatccctg ctgggcaaga
     1081 tctgcgatct aagtaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc
     1141 caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga
     1201 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
     1261 tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc
     1321 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
     1381 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
     1441 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac
     1501 cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc
     1561 aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
     1621 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga
     1681 gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct
     1741 gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga
     1801 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     1861 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     1921 aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag
     1981 tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata
     2041 cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg
     2101 ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga
     2161 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     2221 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     2281 taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa
     2341 tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc
     2401 ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta
     2461 caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat
     2521 cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt
     2581 tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag
     2641 tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa
     2701 aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa
     2761 gggcggaaag atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat
     2821 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2881 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2941 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3001 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga
     3061 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     3121 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     3181 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3241 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3301 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3361 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3421 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3481 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3541 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3601 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3661 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3721 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3781 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3841 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3901 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3961 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4021 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4081 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4141 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4201 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4261 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4321 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4381 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4441 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4501 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4561 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4621 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4681 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4741 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4801 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4861 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4921 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4981 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5041 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5101 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5161 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5221 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5281 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5341 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5401 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5461 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5521 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca
     5581 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5641 cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca
     5701 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     5761 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     5821 ccagtgcaag tgcaggtgcc agaacatttc tctatcgata
//
