LOCUS       sequence-331443-        9141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NEK9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9141)
  AUTHORS   Yamamoto Y, Chino H, Tsukamoto S, Ode KL, Ueda HR, Mizushima N
  TITLE     NEK9 regulates primary cilia formation by acting as a selective 
            autophagy adaptor for MYH9/myosin IIA.
  JOURNAL   Nat Commun. 2021 Jun 2;12(1):3292. doi: 10.1038/s41467-021-23599-7.
  PUBMED    34078910
REFERENCE   2  (bases 1 to 9141)
  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: "2021-06-2"; volume: "12(1):3292. doi"; pages: " 
            10.1038/s41467-021-23599-7"
COMMENT     Sequence Label: pMRX-IPU-FLAG-NEK9
FEATURES             Location/Qualifiers
     source          1..9141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(272..289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(443..1031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(523..542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1195..1986,1987..2055))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1818..1837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2056..2160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2228..2246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2348..2727
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    3086..3443
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3508..3924
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3854..3876
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3892..3908
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3934..4308
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     4246..4262
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      4335..4344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4344..4409
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6317..6328
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    7399..7965
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7415..7967
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7559..7576)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7786..7805
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7985..8584
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7985..8004)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8481..8501
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             join(8814..9141,1..142)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
       61 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
      121 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
      181 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
      241 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
      301 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      361 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      421 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      481 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      901 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      961 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1021 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1081 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1141 tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1801 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2161 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2221 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac tataggagct tggcccattg
     2281 catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc aacattaccg
     2341 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     2401 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2461 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2521 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2581 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2641 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2701 tacgtattag tcatcgctat taccatgcta gagaaccatc agatgtttcc agggtgcccc
     2761 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     2821 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     2881 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     2941 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     3001 cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg
     3061 gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt
     3121 gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct aactagctct
     3181 gtatctggcg gacccgtggt ggaactgacg agttcggaac acccggccgc aaccctggga
     3241 gacgtcccag ggacttcggg ggccgttttt gtggcccgac ctgagtccaa aaatcccgat
     3301 cgttttggac tctttggtgc acccccctta gaggagggat atgtggttct ggtaggagac
     3361 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg ggaccgaagc
     3421 cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
     3481 ctgtatttgt ctgaaaatat gggcccgggc cagactgtta ccactccctt aagtttgacc
     3541 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     3601 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     3661 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     3721 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     3781 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     3841 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     3901 gccctcactc cttctctagg cgcccccata tggccatatg agatcttata tggggcaccc
     3961 ccgccccttg taaacttccc tgaccctgac atgacaagag ttactaacag cccctctctc
     4021 caagctcact tacaggctct ctacttagtc cagcacgaag tctggagacc tctggcggca
     4081 gcctaccaag aacaactgga ccgaccggtg gtacctcacc cttaccgagt cggcgacaca
     4141 gtgtgggtcc gccgacacca gactaagaac ctagaacctc gctggaaagg accttacaca
     4201 gtcctgctga ccacccccac cgccctcaaa gtagacggca tcgcagcttg gatacacgcc
     4261 gcccacgtga aggctgccga ccccgggggt ggaccatcct ctagactgcc ggatctgcca
     4321 tcgatataaa gcttgccacc atggactaca aagaccatga cggtgattat aaagatcatg
     4381 acatcgacta caaggatgac gatgacaagg gttcaggctc gggatcctcg gtgctgggcg
     4441 agtacgagcg acactgcgat tccatcaact cggactttgg gagcgagtcc gggggttgcg
     4501 gggactcgag tccggggcct agcgccagtc aggggccgcg agccggcggc ggcgcggcgg
     4561 agcaggagga actgcactac atccccatcc gcgtcctggg ccgcggcgcc ttcggggaag
     4621 ccacgctgta ccgccgcacc gaggatgact cactggttgt gtggaaggaa gtcgatttga
     4681 cccggctgtc tgagaaggaa cgtcgtgatg ccttgaatga gatagttatt ctggcactgc
     4741 tgcagcacga caacattatt gcctactaca atcacttcat ggacaatacc acgctgctga
     4801 ttgagctgga atattgtaat ggagggaacc tgtatgacaa aatccttcgt cagaaggaca
     4861 agttgtttga ggaagagatg gtggtgtggt acctatttca gattgtttca gcagtgagct
     4921 gcatccataa agctggaatc cttcatagag atataaagac attaaatatt tttctgacca
     4981 aggcaaacct gataaaactt ggagattatg gcctagcaaa gaaacttaat tctgagtatt
     5041 ccatggctga gacgcttgtg ggaaccccat attacatgtc tccagagctc tgtcaaggag
     5101 taaagtacaa tttcaagtct gatatctggg cagttggctg cgtcattttt gaactgctta
     5161 ccttaaagag gacgtttgat gctacaaacc cacttaacct gtgtgtgaag atcgtgcaag
     5221 gaattcgggc catggaagtt gactctagcc agtactcttt ggaattgatc caaatggttc
     5281 attcgtgcct tgaccaggat cctgagcaga gacctactgc agatgaactt ctagatcgcc
     5341 ctcttctcag gaaacgcagg agagagatgg aggaaaaagt cactctgctt aatgcaccta
     5401 caaagagacc aaggtcaagc actgtgactg aagcacccat tgctgtagta acatcacgaa
     5461 ccagtgaagt ctatgtttgg ggtggtggaa aatccacccc ccagaaactg gatgttatca
     5521 agagtggctg tagtgcccgg caggtctgtg cagggaatac ccactttgct gtggtcacag
     5581 tggagaagga actgtacact tgggtgaaca tgcaaggagg cactaaactc catggtcagc
     5641 tgggccatgg agacaaagcc tcctatcgac agccaaagca tgtggaaaag ttgcaaggca
     5701 aagctatccg tcaggtgtca tgtggtgatg atttcactgt ctgtgtgact gatgagggtc
     5761 agctctatgc cttcggatca gattattatg gctgcatggg ggtggacaaa gttgctggcc
     5821 ctgaagtgct agaacccatg cagctgaact tcttcctcag caatccagtg gagcaggtct
     5881 cctgtggaga taatcatgtg gtggttctga cacgaaacaa ggaagtctat tcttggggct
     5941 gtggcgaata tggacgactg ggtttggatt cagaagagga ttattataca ccacaaaagg
     6001 tggatgttcc caaggccttg attattgttg cagttcaatg tggctgtgat gggacatttc
     6061 tgttgaccca gtcaggcaaa gtgctggcct gtggactcaa tgaattcaat aagctgggtc
     6121 tgaatcagtg catgtcggga attatcaacc atgaagcata ccatgaagtt ccctacacaa
     6181 cgtcctttac cttggccaaa cagttgtcct tttataagat ccgtaccatt gccccaggca
     6241 agactcacac agctgctatt gatgagcgag gccggctgct gacctttggc tgcaacaagt
     6301 gtgggcagct gggcgttggg aactacaaga agcgtctggg aatcaacctg ttggggggac
     6361 cccttggtgg gaagcaagtg atcagggtct cctgcggtga tgagtttacc attgctgcca
     6421 ctgatgataa tcacattttt gcctggggca atggtggtaa tggccgcctg gcaatgaccc
     6481 ccacagagag accacatggc tctgatatct gtacctcatg gcctcggcct atttttggat
     6541 ctctgcatca tgtcccggac ctgtcttgcc gtggatggca taccattctc atcgttgaga
     6601 aagtattgaa ttctaagacc atccgttcca atagcagtgg cttatccatt ggaactgtgt
     6661 ttcagagctc tagcccggga ggaggcggcg ggggcggcgg tggtgaagaa gaggacagtc
     6721 agcaggaatc tgaaactcct gacccaagtg gaggcttccg aggaacaatg gaagcagacc
     6781 gaggaatgga aggtttaatc agtcccacag aggccatggg gaacagtaat ggggccagca
     6841 gctcctgtcc tggctggctt cgaaaggagc tggaaaatgc agaatttatc cccatgcctg
     6901 acagcccatc tcctctcagt gcagcgtttt cagaatctga gaaagatacc ctgccctatg
     6961 aagagctgca aggactcaaa gtggcctctg aagctccttt ggaacacaaa ccccaagtag
     7021 aagcctcgtc acctcggctg aatcctgcag taacctgtgc tgggaaggga acaccactga
     7081 ctcctcctgc gtgtgcgtgc agctctctgc aggtggaggt tgagagattg cagggtctgg
     7141 tgttaaagtg tctggctgaa caacagaagc tacagcaaga aaacctccag atttttaccc
     7201 aactgcagaa gttgaacaag aaattagaag gagggcagca ggtggggatg cattccaaag
     7261 gaactcagac agcaaaggaa gagatggaaa tggatccaaa gcctgactta gattcagatt
     7321 cctggtgcct cctgggaaca gactcctgta gacccagcct ctaggaattc gggtctcgag
     7381 agcggccgct acgtaaattc cgcccccccc cctaacgtta ctggccgaag ccgcttggaa
     7441 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     7501 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     7561 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagtt
     7621 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     7681 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     7741 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     7801 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     7861 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     7921 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccgt
     7981 cgacatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     8041 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     8101 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     8161 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     8221 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     8281 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     8341 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     8401 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     8461 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     8521 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     8581 ctgagtcgac cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
     8641 tggctcgacg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     8701 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     8761 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     8821 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     8881 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     8941 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     9001 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     9061 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     9121 cccacaaccc ctcactcggg g
//
