LOCUS       sequence-265893-        5626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter activity of the CLEAR network.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5626)
  AUTHORS   Shin HR, Kim H, Oh S, Lee JG, Kee M, Ko HJ, Kweon MN, Won KJ, Baek 
            SH
  TITLE     AMPK-SKP2-CARM1 signalling cascade in transcriptional regulation of 
            autophagy.
  JOURNAL   Nature. 2016 Jun 15. doi: 10.1038/nature18014.
  PUBMED    27309807
REFERENCE   2  (bases 1 to 5626)
  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: "Nature. 
            2016 Jun 15. doi: 10.1038/nature18014."
COMMENT     Sequence Label: pGL2-2xCLEAR-luciferase
FEATURES             Location/Qualifiers
     source          1..5626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    646..767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             896..2548
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(962..980)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2789..2854
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2984..3004
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3429..3563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3705..3722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3876..4464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3956..3975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4635..5426,5427..5495))
                     /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     5258..5277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5496..5600)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgtggag gttttacttg ctttaaaaaa cctcccacac
      601 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
      661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
      721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatgg tactgtaact
      781 gagctaacat aacccgggag gtaccgagct cttaccgggt cacgtgaccc cagggtcacg
      841 tgaccctgct agctcgagat ctaagtaagc ttggcattcc ggtactgttg gtaaaatgga
      901 agacgccaaa aacataaaga aaggcccggc gccattctat cctctagagg atggaaccgc
      961 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
     1021 tacagatgca catatcgagg tgaacatcac gtacgcggaa tacttcgaaa tgtccgttcg
     1081 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
     1141 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
     1201 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatga acatttcgca
     1261 gcctaccgta gtgtttgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
     1321 attaccaata atccagaaaa ttattatcat ggattctaaa acggattacc agggatttca
     1381 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
     1441 accagagtcc tttgatcgtg acaaaacaat tgcactgata atgaattcct ctggatctac
     1501 tgggttacct aagggtgtgg cccttccgca tagaactgcc tgcgtcagat tctcgcatgc
     1561 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     1621 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     1681 cgtcttaatg tatagatttg aagaagagct gtttttacga tcccttcagg attacaaaat
     1741 tcaaagtgcg ttgctagtac caaccctatt ttcattcttc gccaaaagca ctctgattga
     1801 caaatacgat ttatctaatt tacacgaaat tgcttctggg ggcgcacctc tttcgaaaga
     1861 agtcggggaa gcggttgcaa aacgcttcca tcttccaggg atacgacaag gatatgggct
     1921 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     1981 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     2041 gggcgttaat cagagaggcg aattatgtgt cagaggacct atgattatgt ccggttatgt
     2101 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     2161 catagcttac tgggacgaag acgaacactt cttcatagtt gaccgcttga agtctttaat
     2221 taaatacaaa ggatatcagg tggcccccgc tgaattggaa tcgatattgt tacaacaccc
     2281 caacatcttc gacgcgggcg tggcaggtct tcccgacgat gacgccggtg aacttcccgc
     2341 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     2401 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     2461 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     2521 caagaagggc ggaaagtcca aattgtaaaa tgtaactgta ttcagcgatg acgaaattct
     2581 tagctattgt aatactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     2641 tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc
     2701 agaaattcgc cggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa
     2761 actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt
     2821 taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg
     2881 ggagcagtgg tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc
     2941 tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa
     3001 ggtagaagac cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt
     3061 tagtaataga actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct
     3121 atacaagaaa attatggaaa aatattctgt aacctttata agtaggcata acagttataa
     3181 tcataacata ctgttttttc ttactccaca caggcataga gtgtctgcta ttaataacta
     3241 tgctcaaaaa ttgtgtacct ttagcttttt aatttgtaaa ggggttaata aggaatattt
     3301 gatgtatagt gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac
     3361 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3421 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3541 atgtatctta tcatgtctgg atccgtcgac cgatgccctt gagagccttc aacccagtca
     3601 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     3661 tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg
     3721 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3781 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3841 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3901 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3961 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4021 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4081 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4141 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4201 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4261 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4321 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4381 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4441 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4501 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4561 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4621 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4681 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4741 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4801 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4861 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4921 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4981 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5041 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5101 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5161 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5221 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5281 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5341 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5401 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5461 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5521 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5581 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
