LOCUS       sequence-178509-        6533 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  shRNA against AMPK alpha2 and alpha1 in mouse/human to knock down 
            protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6533)
  AUTHORS   Faubert B, Boily G, Izreig S, Griss T, Samborska B, Dong Z, Dupuy F,
            Chambers C, Fuerth BJ, Viollet B, Mamer OA, Avizonis D, DeBerardinis
            RJ, Siegel PM, Jones RG
  TITLE     AMPK is a negative regulator of the Warburg effect and suppresses 
            tumor growth in vivo.
  JOURNAL   Cell Metab. 2013 Jan 8;17(1):113-24. doi: 
            10.1016/j.cmet.2012.12.001. Epub 2012 Dec 27.
  PUBMED    23274086
REFERENCE   2  (bases 1 to 6533)
  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"; doi: 
            "10.1016/j.cmet.2012.12.001"; journalName: "Cell Metab"; date: 
            "2013-01-8- 8"; volume: "17"; issue: "1"; pages: "113-24"
COMMENT     Sequence Label: MSCV p2GM AMPK alpha2hp1 alpha1hp1
FEATURES             Location/Qualifiers
     source          1..6533
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     343..363
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             474..539
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(549..551,552..554,555..1268)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(549..551,552..554,555..1265)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(594..615)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(855..874)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1202..1223
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           1340..1461
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1489..1503
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1531..1657
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1734..1855
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1883..1897
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1925..2051
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    2108..2696
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2161..2181)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2579..2590)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2743..3257
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3419..3435
                     /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(3443..3449,3450..3467,3468..3473))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3488..3509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3626..3643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3797..4385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3877..3896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4556..5347,5348..5416))
                     /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     5179..5198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5417..5521)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5589..5607
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5645..5667)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5767..5786
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5980..6002
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6130..6149
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
       61 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
      121 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
      181 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
      241 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
      301 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
      361 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
      421 gcatgacccg caagcccggt gccctgtaca agaaacagaa aattgtggca ccagtgaaac
      481 agactttgaa ttttgacctt ctcaagttgg cgggagacgt cgagtccaac cctggcccgg
      541 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      601 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      661 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      721 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      781 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      841 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      901 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      961 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1021 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1081 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1141 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1201 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1261 acaagtagcg gccgcagcag cagcatttaa atggccgcaa gccttgttaa gtgctcgctt
     1321 cggcagcaca tatactatgt tgaatgaggc ttcagtactt tacagaatcg ttgcctgcac
     1381 atcttggaaa cacttgctgg gattacttct tcaggttaac ccaacagaag gctcgagaag
     1441 gtatattgct gttgacagtg agcgcggcca taaagtggca gttaaatagt gaagccacag
     1501 atgtatttaa ctgccacttt atggccttgc ctactgcctc ggaattcaag gggctacttt
     1561 aggagcaatt atcttgttta ctaaaactga ataccttgct atctctttga tacattttta
     1621 caaagctgaa ttaaaatggt ataaattaaa tcactttttt caattggaag actaatgcgt
     1681 ttaaatggcc gcaagccttg ttaagtgctc gcttcggcag cacatatact atgttgaatg
     1741 aggcttcagt actttacaga atcgttgcct gcacatcttg gaaacacttg ctgggattac
     1801 ttcttcaggt taacccaaca gaaggctcga gaaggtatat tgctgttgac agtgagcgcg
     1861 gaggagagct atttgattat tagtgaagcc acagatgtaa taatcaaata gctctcctcc
     1921 atgcctactg cctcggaatt caaggggcta ctttaggagc aattatcttg tttactaaaa
     1981 ctgaatacct tgctatctct ttgatacatt tttacaaagc tgaattaaaa tggtataaat
     2041 taaatcactt ttttcaattg gaagactaat gcgtttaaac tgtcgacctg cagccaagct
     2101 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2161 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2221 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2281 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2341 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2401 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2461 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2521 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2581 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2641 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     2701 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     2761 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga
     2821 gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag
     2881 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     2941 gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     3001 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     3061 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     3121 tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     3181 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     3241 cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta
     3301 ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca
     3361 atactcctga aatagttcat tatggacagc gcagaaagag ctggggagaa ttgtgaaatt
     3421 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3481 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3541 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3601 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3661 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3721 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3781 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3841 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3901 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3961 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4021 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4081 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4141 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4201 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4261 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4321 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4381 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4441 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4501 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4561 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4621 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4681 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4741 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4801 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4861 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4921 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4981 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5041 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5101 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5161 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5221 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5281 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5341 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5401 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5461 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5521 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5581 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5641 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5701 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5761 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     5821 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     5881 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     5941 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     6001 cgacggcgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     6061 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     6121 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     6181 gccggccacg atgcgtccgg cgtagaggcg attagtccaa tttgttaaag acaggatatc
     6241 agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga
     6301 gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     6361 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
     6421 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     6481 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga aca
//
