LOCUS       sequence-379143-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mScarlet-tagged alpha2 isoform of AMPK catalytic subunit targeted to
            the nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  AUTHORS   Schmitt DL, Curtis SD, Lyons AC, Zhang JF, Chen M, He CY, Mehta S, 
            Shaw RJ, Zhang J
  TITLE     Spatial regulation of AMPK signaling revealed by a sensitive kinase 
            activity reporter.
  JOURNAL   Nat Commun. 2022 Jul 5;13(1):3856. doi: 10.1038/s41467-022-31190-x.
  PUBMED    35790710
REFERENCE   2  (bases 1 to 6415)
  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: "Nat 
            Commun."; date: "2022-07-5"; volume: "13(1):3856. doi"; pages: " 
            10.1038/s41467-022-31190-x"
COMMENT     Sequence Label: pEGFP-C1-3x-NLS-mScarlet-AMPK-alpha-2
FEATURES             Location/Qualifiers
     source          1..6415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        141..444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        445..648
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     599..619
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             711..731
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             735..755
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             759..779
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             789..1484
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(922..940)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1190..1209
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3355..3476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3392..3411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3446..3465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3483..3938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3620..3641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3832..3851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3965..4069
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4094..4114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4138..4348
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4160..4356
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4207..4342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4391..5185
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4391..5185
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4445..4464)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5055..5074
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5373..5392)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5417..5464
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5793..6381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6282..6301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
       61 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
      121 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      181 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      241 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      301 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      361 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      421 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      481 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      541 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      601 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      661 cgtcagatcc gctagcgcta ccggtcgcca ccatgagatc tcgagctgat ccaaaaaaga
      721 agagaaaggt agatccaaaa aagaagagaa aggtagatcc aaaaaagaag agaaaggtag
      781 gatccagtat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
      841 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      901 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
      961 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
     1021 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     1081 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     1141 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     1201 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     1261 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     1321 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     1381 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     1441 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccggc cggactcaga
     1501 tctcgagctc aagcttcgaa ttcaacatgg ctgagaagca gaagcacgac gggcgtgtga
     1561 agatcggaca ctacgtgctg ggggacaccc tgggcgtcgg caccttcggc aaagtgaaga
     1621 ttggagaaca tcaattgaca ggccataaag tggcagttaa gatcttaaat agacagaaga
     1681 ttcgcagttt agatgttgtt ggaaaaataa aacgagaaat tcaaaatctt aaactctttc
     1741 gtcatcctca tattatcaaa ctctaccaag tgatcagcac tccaacagac ttttttatgg
     1801 taatggaata tgtgtctgga ggtgaattgt tcgactacat ctgtaaacac gggagggttg
     1861 aagaggtgga agctcgccgg ctcttccagc agattctgtc tgccgtggac tactgtcaca
     1921 ggcacatggt tgtccacagg gacctgaagc cagagaacgt gttgctggac gcccagatga
     1981 atgctaagat agctgacttc ggactctcta atatgatgtc agatggtgaa tttctacgaa
     2041 ctagctgtgg atcgccaaat tatgcagcac cggaggtcat ctcaggaagg ctgtatgcgg
     2101 gtcctgaggt tgatatctgg agctgtggtg ttatcctgta tgcccttctc tgtggcaccc
     2161 tcccgttcga cgatgagcac gtgcctacgc tctttaagaa gatccgaggg ggtgtgttct
     2221 acatcccgga gtatctcaac cgttctattg ccactctgct gatgcacatg ctgcaggtgg
     2281 accccttgaa gcgagcaact atcaaagaca tacgagagca tgaatggttt aaacaggatt
     2341 tgcccagtta cctctttcct gaagacccct cctatgatgc taacgtcatt gatgatgagg
     2401 ctgtgaaaga agtatgtgaa aaatttgagt gtacagaatc agaagtgatg aacagtttat
     2461 acagtggtga ccctcaagac cagctcgcag tggcttatca tctcatcatt gacaatcgga
     2521 gaataatgaa ccaagccagt gagttctacc tcgcctccag tcctccaacg ggttccttca
     2581 tggacgatag tgccatgcat attccccccg gcctgaaacc acatcctgaa aggatgccac
     2641 ctctcatagc agacagcccc aaagcacgct gtccactgga tgcactcaac acaactaagc
     2701 ccaaatcttt agctgtgaaa aaagccaagt ggcaccttgg gatccgaagc cagagcaaac
     2761 catacgacat tatggcggag gtgtaccgag ctatgaagca gctggacttt gaatggaagg
     2821 tagtgaatgc ataccatctt cgagtaagaa gaaaaaaccc agtgactggc aattacgtga
     2881 aaatgagctt acagctttac ctggttgaca atcggagcta tcttctagac tttaaaagca
     2941 tcgatgatga ggtggtggag cagaggtctg gttcttcaac acctcagcgc tcctgttctg
     3001 ctgccggcct ccacagacct cggtcaagtg tcgattccag cacagccgag aaccattcac
     3061 tgtctggctc tctcactggt tctttgactg gcagcacttt gtcctccgct tccccgcgcc
     3121 tgggcagtca taccatggat ttttttgaaa tgtgcgccag tcttatcact gctttagccc
     3181 gttgaggatc cataaagggc gaattccaca ttgggctgca gcccgggatc caccggatct
     3241 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3301 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3481 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3541 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3601 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3661 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3721 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3781 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3841 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3901 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4081 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4201 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4261 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4321 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4381 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4441 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4501 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4561 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4621 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4681 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4741 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4801 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4861 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4921 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4981 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5041 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5101 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5161 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5221 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5281 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5341 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5401 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5461 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5521 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5581 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5641 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5701 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5761 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5821 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5881 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5941 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6001 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6061 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctgg
//
