LOCUS       sequence-176882-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes C-terminal (substrate) fragment of bimolecular AMPK/BRSK 
            activity reporter (bimABKAR); plasma membrane targeted; use in 
            conjunction with pcDNA3-Cerulean-FHA1-NES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  AUTHORS   Depry C, Mehta S, Li R, Zhang J
  TITLE     Visualization of Compartmentalized Kinase Activity Dynamics Using 
            Adaptable BimKARs.
  JOURNAL   Chem Biol. 2015 Nov 19;22(11):1470-9. doi: 
            10.1016/j.chembiol.2015.10.004. Epub 2015 Nov 5.
  PUBMED    26548610
REFERENCE   2  (bases 1 to 6290)
  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.chembiol.2015.10.004"; journalName: "Chem Biol"; date: 
            "2015-11-19- 19"; volume: "22"; issue: "11"; pages: "1470-9"
COMMENT     Sequence Label: pcDNA3-AMPKsub-YPet-Kras
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             22..54
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(58..66,67..81)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(448..467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(922..940)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(923..940)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(960..977)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    966..1190
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1236..1664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1323..1342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1533..1554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1673..1693)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1678..2007
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1858..1993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1920..1939
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2074..2868
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2128..2147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2738..2757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3042..3163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3079..3098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3133..3152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3212..3228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3212..3228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3225..3247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3236..3252
                     /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(3260..3266,3267..3284,3285..3290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3305..3326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3443..3460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3614..4202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3694..3713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4373..5164,5165..5233))
                     /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     4996..5015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5234..5338)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5413..5432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5604..5983
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5984..6187
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6138..6158
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6232..6251
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6232..6250
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6270..6279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
       61 ctgtacgacg atgacgataa ggatcccatg aggagagtgg ctaccctggt ggatctgggc
      121 ggcaccggcg gcagcgagct catggtgagc aaaggcgaag agctgttcac cggcgtggtg
      181 cccatcctgg tggagctgga cggcgacgtg aacggccaca agttcagcgt gagcggcgag
      241 ggcgagggcg acgccaccta cggcaagctg accctgaagc tgctgtgcac caccggcaag
      301 ctgcccgtgc cctggcccac cctggtgacc accctgggct acggcgtgca gtgcttcgcc
      361 cggtaccccg accacatgaa gcagcacgac ttcttcaaga gcgccatgcc cgagggctac
      421 gtgcaggagc ggaccatctt cttcaaggac gacggcaact acaagacccg ggccgaggtg
      481 aagttcgagg gcgacaccct ggtgaaccgg atcgagctga agggcatcga cttcaaggag
      541 gacagcaaca tcctgggcca caagctggag tacaactaca acagccacaa cgtgtacatc
      601 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccggca caacatcgag
      661 gacggcggcg tgcagctggc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      721 gtgctgctgc ccgataacca ctacctgagc taccagagcg ccctgttcaa ggaccccaac
      781 gagaagcggg accacatggt gctgctggag ttcctgaccg ccgccggcat caccgagggt
      841 atgaatgaat tgtacaaaga attcaagaag aagaagaaga gcaagaccaa gtgcgtgatc
      901 atgtaatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc
      961 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1021 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1081 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1141 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     1201 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     1261 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1321 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1381 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1441 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1501 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1561 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1621 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     1681 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     1741 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     1801 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     1861 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     1921 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     1981 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     2041 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2101 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2161 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2221 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     2281 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2341 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2401 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2461 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     2521 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     2581 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     2641 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     2701 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     2761 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     2821 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     2881 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     2941 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3001 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3061 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     3181 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3301 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3361 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4021 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5341 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     5401 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     5461 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     5521 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     5581 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     5641 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5701 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5761 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5821 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5881 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     5941 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     6001 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6061 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6121 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6181 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     6241 tcactatagg gagacccaag cttgcggccg ccaccatggt gcggggttct
//
