LOCUS       sequence-329932-        8188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of Nuak1 (AMPK-related kinase also called 
            ARK5/Omphk1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8188)
  AUTHORS   Courchet J, Lewis TL Jr, Lee S, Courchet V, Liou DY, Aizawa S, 
            Polleux F
  TITLE     Terminal axon branching is regulated by the LKB1-NUAK1 kinase 
            pathway via presynaptic mitochondrial capture.
  JOURNAL   Cell. 2013 Jun 20;153(7):1510-25. doi: 10.1016/j.cell.2013.05.021.
  PUBMED    23791179
REFERENCE   2  (bases 1 to 8188)
  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.cell.2013.05"; journalName: "Cell"; date: "2013-06-20- 
            20"; volume: "153"; issue: "7"; pages: "1510-25"
COMMENT     Sequence Label: pCIG2-Flag-mNuak1 WT
FEATURES             Location/Qualifiers
     source          1..8188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..685
                     /label=chicken beta-actin promoter
     intron          686..1702
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1626..1648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1710..1729
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1766..1789
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(3805..4379,4380..4382,4383..4391)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3972..3989)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4199..4218
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4392..4394,4395..4397,4398..5111)
                     /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(4392..4394,4395..4397,4398..5108)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4437..4458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4698..4717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5045..5066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5203..5222)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5268..5323
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5322..5341)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5713..5729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5713..5729)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5726..5748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5737..5753
                     /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(5761..5767,5768..5785,5786..5791))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5806..5827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5886..6081
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5932..6067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6087..6221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6124..6143)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6178..6197
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6289..6306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6460..7048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6540..6559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7219..8010,8011..8079))
                     /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     7842..7861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8080..8184)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      421 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      481 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccrgg
      541 sggggsgggg sggggsgrgg ggsggggsgg ggcgaggcgg agaggtgcgg cggcagccaa
      601 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      661 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      721 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      781 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
      841 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
      901 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
      961 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     1021 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     1081 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     1141 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     1201 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     1261 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     1321 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     1381 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     1441 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     1501 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     1561 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     1621 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     1681 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     1741 ttttggcaaa gaattgctcg agatggatta caaggatgac gacgataaga tggaaggggc
     1801 ggcagtgtcc gcggcagggg atggccctgc tgtagagacc ggcttgccgg gctctccgct
     1861 ggaggcggtg gccggggcga ctgcagcccc agtggagccc aggaagccgc acggggtgaa
     1921 gcgacatcac cacaaacaca acttgaagca tcgctatgag ctgcaagaga ccctgggcaa
     1981 aggtacctac ggcaaagtca agagagccac cgagaggttt tcaggccgag tggttgctat
     2041 aaaatccatc cgtaaggaca aaattaagga cgagctagac atggttcaca tcagacgaga
     2101 gattgagatc atgtcatccc tcaaccatcc tcatatcatc agtatttatg aagtgtttga
     2161 gaacaaagat aagattgtga tcatcatgga atatgccagc aaaggagagc tgtatgatta
     2221 catcagtgag cgcaggcgcc tcagcgagag ggagaccaga cacttcttcc ggcagattgt
     2281 gtctgccgtg cattactgtc acaagaatgg tgtggtccac cgggacttga agctggagaa
     2341 catactgctt gatgacaact gcaacattaa gattgctgac tttgggctct ccaacctgta
     2401 ccagaaggac aagttcttgc aaacattttg tgggagccca ctctatgcgt cgcccgagat
     2461 tgtcaacggc aggccttacc gagggccaga ggtggacagc tgggccctgg gtgtcttgct
     2521 ttacactctc atttatggaa cgatgccctt cgatggcttt gatcacaaaa acctcatccg
     2581 gcagatcagc agcggagagt accgggagcc cacacaaccc tcagatgctc ggggactcat
     2641 ccggtggatg ctgatggtga atcccgatcg cagagccacc atcgaggaca tcgccaacca
     2701 ttggtgggtg aactggggct acaagagcag cgtctgtgac tgtgatgccc ttcctgactc
     2761 tgagtctcca ctcttggcac ggatcatcga ttggcatcac cgttccacgg ggctgcaggc
     2821 tgaggctgaa gccaagatga agggtctggc caaacccggg gcctctgagg ttgtgttgga
     2881 gcggcagcgg tcactgaaga agtccaagaa agagaacgac tttccccagt ctggccagga
     2941 ctccgtacct gaaagcccat ccaagctgag ttccaagagg cccaagggca tcctgaagaa
     3001 acggagcaac agcgaacacc gctcccacag cactggcttt atcgaaggca ttgtgagccc
     3061 tgccttacca tcccctttca aaatggagca agatttgtgc cggactgcca tccccctccc
     3121 cagctccccc gaggcagata tgtcagggaa gctcagcctc aaacagtctg ccacgatgcc
     3181 caagaaaggc atcttgaaaa agacccagca gagagaatct ggctactatt catccccaga
     3241 gcgcagcgag tcctcggaac tgctggacag caacgatgtg gtcatcagtg gtggcctctc
     3301 gtcccctccc ccagacccag ccaggggaac ttcccacagc ctgtcctgcc ggaggaaggg
     3361 catcttaaaa catagtagca ggtactcaga tggtggcaca gaccctgccc tcacccgccc
     3421 tgaaatgccc acactggaat ccttgtcacc tcccggtgtc ccctccgatg gcatctctcg
     3481 gagctacagt cgcccgtcca gtatcatcag tgatgacagt gtcctgtcca gcgactcttt
     3541 cgacttgtta gagcttcaag agaaccggcc agcccgccag cgcatccgca gctgcgtctc
     3601 tgctgaaaac ttcctccagc ttcaggactt tgagacacca cacaaccggc cccggcccca
     3661 gtacctgaag cggctggcag acagtagctt ctcccttctc actgatatgg acgatgtgac
     3721 tcaggtgtat aagaaagcgc tggagatctg cagcaagctc aactaggaat tctgcagtcg
     3781 acggtaccgc gggcccggga tccgcccctc tccctccccc ccccctaacg ttactggccg
     3841 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     3901 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     3961 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     4021 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     4081 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     4141 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     4201 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     4261 gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa
     4321 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     4381 tggccacaac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     4441 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     4501 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     4561 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     4621 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     4681 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     4741 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     4801 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     4861 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     4921 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     4981 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     5041 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     5101 tgtacaagta aagcggccgc aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     5161 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     5221 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     5281 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     5341 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5401 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     5461 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     5521 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     5581 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     5641 atggagatcc ctcgacctgc accgtcgacc agcttggcgw mmtsmwssky mkascwgctt
     5701 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5761 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5821 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     5881 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6001 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6061 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     6121 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6181 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     6241 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6301 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6361 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6421 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6481 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6541 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6601 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6661 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6721 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6781 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6841 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6901 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6961 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7021 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7081 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7141 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7201 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7261 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7321 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7381 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7441 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7501 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7561 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7621 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7681 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7741 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7801 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7861 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7921 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7981 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8041 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8101 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8161 aaataaacaa ataggggttc cgcgcaca
//
