LOCUS       sequence-176148-        7508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry-tagged, LKB1(K78I); kinase-dead mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7508)
  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 7508)
  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-mCherry-LKB1(KD)
FEATURES             Location/Qualifiers
     source          1..7508
                     /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"
     CDS             88..795
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(233..251)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     501..520
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(2140..2158)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2141..2158)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2178..2195)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2184..2408
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2454..2882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2541..2560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2751..2772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2891..2911)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2896..3225
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3076..3211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3138..3157
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3292..4086
                     /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(3346..3365)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3956..3975
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4260..4381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4297..4316)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4351..4370
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4430..4446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4430..4446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4443..4465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4454..4470
                     /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(4478..4484,4485..4502,4503..4508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4523..4544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4661..4678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4832..5420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4912..4931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5591..6382,6383..6451))
                     /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     6214..6233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6452..6556)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6631..6650)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6822..7201
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7202..7405
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7356..7376
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7450..7469
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7450..7468
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      7488..7497
                     /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 gtgagcaagg gcgaggagga taacatggcc
      121 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      181 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      241 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
      301 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
      361 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
      421 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
      481 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
      541 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
      601 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
      661 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
      721 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
      781 gacgagctgt acaaggaatt ctccagcatg gaggtggtgg acccgcagca gctgggcatg
      841 ttcacggagg gcgagctgat gtcggtgggt atggacacgt tcatccaccg catcgactcc
      901 accgaggtca tctaccagcc gcgccgcaag cgggccaagc tcatcggcaa gtacctgatg
      961 ggggacctgc tgggggaagg ctcttacggc aaggtgaagg aggtgctgga ctcggagacg
     1021 ctgtgcagga gggccgtcat catcctcaag aagaagaagt tgcgaaggat ccccaacggg
     1081 gaggccaacg tgaagaagga aattcaacta ctgaggaggt tacggcacaa aaatgtcatc
     1141 cagctggtgg atgtgttata caacgaagag aagcagaaaa tgtatatggt gatggagtac
     1201 tgcgtgtgtg gcatgcagga aatgctggac agcgtgccgg agaagcgttt cccagtgtgc
     1261 caggcccacg ggtacttctg tcagctgatt gacggcctgg agtacctgca tagccagggc
     1321 attgtgcaca aggacatcaa gccggggaac ctgctgctca ccaccggtgg caccctcaaa
     1381 atctccgacc tgggcgtggc cgaggcactg cacccgttcg cggcggacga cacctgccgg
     1441 accagccagg gctccccggc tttccagccg cccgagattg ccaacggcct ggacaccttc
     1501 tccggcttca aggtggacat ctggtcggct ggggtcaccc tctacaacat caccacgggt
     1561 ctgtacccct tcgaagggga caacatctac aagttgtttg agaacatcgg gaaggggagc
     1621 tacgccatcc cgggcgactg tggccccccg ctctctgacc tgctgaaagg gatgcttgag
     1681 tacgaaccgg ccaagaggtt ctccatccgg cagatccggc agcacagctg gttccggaag
     1741 aaacatcctc cggctgaagc accagtgccc atcccaccga gcccagacac caaggaccgg
     1801 tggcgcagca tgactgtggt gccgtacttg gaggacctgc acggcgcgga cgaggacgag
     1861 gacctcttcg acatcgagga tgacatcatc tacactcagg acttcacggt gcccggacag
     1921 gtcccagaag aggaggccag tcacaatgga cagcgccggg gcctccccaa ggccgtgtgt
     1981 atgaacggca cagaggcggc gcagctgagc accaaatcca gggcggaggg ccgggccccc
     2041 aaccctgccc gcaaggcctg ctccgccagc agcaagatcc gccggctgtc ggcctgcaag
     2101 cagcagtgag gcgtcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     2161 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2221 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2281 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2341 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2401 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2461 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2521 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2581 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2641 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2701 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2761 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2821 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2881 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3001 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3241 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3301 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3361 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3421 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3481 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3541 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3601 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3661 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3721 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3781 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3841 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3901 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3961 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4021 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4081 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4141 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4201 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4381 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4441 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4501 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4621 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5341 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5401 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5461 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5521 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5581 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5641 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5701 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5761 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5821 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5881 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5941 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6001 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6061 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6121 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6181 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6241 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6301 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6361 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6421 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6481 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6541 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6601 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6661 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     6721 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     6781 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     6841 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     6901 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     6961 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     7021 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     7081 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     7141 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     7201 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     7261 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     7321 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     7381 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
     7441 tatcgaaatt aatacgactc actataggga gacccaagct tgcggccgcc accatggtgc
     7501 ggggttct
//
