LOCUS       sequence-379144-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mScarlet-tagged alpha2 isoform of AMPK catalytic subunit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6319)
  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 6319)
  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-mScarlet-AMPK-alpha-2
FEATURES             Location/Qualifiers
     source          1..6319
                     /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"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             693..1388
                     /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(826..844)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1094..1113
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3259..3380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3296..3315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3350..3369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3387..3842)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3524..3545)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3736..3755
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3869..3973
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3998..4018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4042..4252
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4064..4260
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4111..4246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4295..5089
                     /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             4295..5089
                     /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(4349..4368)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4959..4978
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5277..5296)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5321..5368
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5697..6285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6186..6205
                     /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 ccatggtgag caagggcgag gcagtgatca
      721 aggagttcat gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga
      781 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
      841 ccaagggtgg ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct
      901 ccagggcctt caccaagcac cccgccgaca tccccgacta ctataagcag tccttccccg
      961 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc
     1021 aggacacctc cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact
     1081 tccctcctga cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc
     1141 ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg
     1201 acggcggccg ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga
     1261 tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca
     1321 ccgtggtgga acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc
     1381 tgtacaagtc cggccggact cagatctcga gctcaagctt cgaattcaac atggctgaga
     1441 agcagaagca cgacgggcgt gtgaagatcg gacactacgt gctgggggac accctgggcg
     1501 tcggcacctt cggcaaagtg aagattggag aacatcaatt gacaggccat aaagtggcag
     1561 ttaagatctt aaatagacag aagattcgca gtttagatgt tgttggaaaa ataaaacgag
     1621 aaattcaaaa tcttaaactc tttcgtcatc ctcatattat caaactctac caagtgatca
     1681 gcactccaac agactttttt atggtaatgg aatatgtgtc tggaggtgaa ttgttcgact
     1741 acatctgtaa acacgggagg gttgaagagg tggaagctcg ccggctcttc cagcagattc
     1801 tgtctgccgt ggactactgt cacaggcaca tggttgtcca cagggacctg aagccagaga
     1861 acgtgttgct ggacgcccag atgaatgcta agatagctga cttcggactc tctaatatga
     1921 tgtcagatgg tgaatttcta cgaactagct gtggatcgcc aaattatgca gcaccggagg
     1981 tcatctcagg aaggctgtat gcgggtcctg aggttgatat ctggagctgt ggtgttatcc
     2041 tgtatgccct tctctgtggc accctcccgt tcgacgatga gcacgtgcct acgctcttta
     2101 agaagatccg agggggtgtg ttctacatcc cggagtatct caaccgttct attgccactc
     2161 tgctgatgca catgctgcag gtggacccct tgaagcgagc aactatcaaa gacatacgag
     2221 agcatgaatg gtttaaacag gatttgccca gttacctctt tcctgaagac ccctcctatg
     2281 atgctaacgt cattgatgat gaggctgtga aagaagtatg tgaaaaattt gagtgtacag
     2341 aatcagaagt gatgaacagt ttatacagtg gtgaccctca agaccagctc gcagtggctt
     2401 atcatctcat cattgacaat cggagaataa tgaaccaagc cagtgagttc tacctcgcct
     2461 ccagtcctcc aacgggttcc ttcatggacg atagtgccat gcatattccc cccggcctga
     2521 aaccacatcc tgaaaggatg ccacctctca tagcagacag ccccaaagca cgctgtccac
     2581 tggatgcact caacacaact aagcccaaat ctttagctgt gaaaaaagcc aagtggcacc
     2641 ttgggatccg aagccagagc aaaccatacg acattatggc ggaggtgtac cgagctatga
     2701 agcagctgga ctttgaatgg aaggtagtga atgcatacca tcttcgagta agaagaaaaa
     2761 acccagtgac tggcaattac gtgaaaatga gcttacagct ttacctggtt gacaatcgga
     2821 gctatcttct agactttaaa agcatcgatg atgaggtggt ggagcagagg tctggttctt
     2881 caacacctca gcgctcctgt tctgctgccg gcctccacag acctcggtca agtgtcgatt
     2941 ccagcacagc cgagaaccat tcactgtctg gctctctcac tggttctttg actggcagca
     3001 ctttgtcctc cgcttccccg cgcctgggca gtcataccat ggattttttt gaaatgtgcg
     3061 ccagtcttat cactgcttta gcccgttgag gatccataaa gggcgaattc cacattgggc
     3121 tgcagcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     3181 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3241 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3301 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3361 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3421 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3481 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3541 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3601 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3661 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3721 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3781 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3841 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3901 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3961 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4141 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4201 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4261 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4321 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4381 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4441 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4501 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4561 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4621 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4681 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4741 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4801 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4861 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4921 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4981 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5041 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5101 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5161 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5221 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5281 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5341 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5401 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5461 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5521 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5581 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5641 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5701 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5761 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5821 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5881 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5941 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6001 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6061 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6121 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6181 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6241 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6301 gcctttttac ggttcctgg
//
