LOCUS       sequence-405567-        8936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xFLAG-tagged NBR1 deltaUBA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8936)
  AUTHORS   Kurusu R, Fujimoto Y, Morishita H, Noshiro D, Takada S, Yamano K, 
            Tanaka H, Arai R, Kageyama S, Funakoshi T, Komatsu-Hirota S, Taka H,
            Kazuno S, Miura Y, Koike M, Wakai T, Waguri S, Noda NN, Komatsu M
  TITLE     Integrated proteomics identifies p62-dependent selective autophagy 
            of the supramolecular vault complex.
  JOURNAL   Dev Cell. 2023 May 9:S1534-5807(23)00191-0. doi: 
            10.1016/j.devcel.2023.04.015.
  PUBMED    37192622
REFERENCE   2  (bases 1 to 8936)
  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: "Dev Cell. 
            2023 May 9:S1534-5807(23)00191-0. doi: 
            10.1016/j.devcel.2023.04.015."
COMMENT     Sequence Label: pMRX-IRES-puro-3xFLAG-NBR1 deltaUBA
FEATURES             Location/Qualifiers
     source          1..8936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..66
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        168..547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    906..1263
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1328..1744
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1674..1696
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1712..1728
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1754..2128
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2066..2082
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2161..2226
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5026..5592
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5042..5594
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5186..5203)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5413..5432
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5606..6205
                     /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(5606..5625)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6102..6122
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6429..6898
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7028..7045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7199..7787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7279..7298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7951..8742,8743..8811))
                     /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     8574..8593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8812..8916)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
       61 cgaggccctt tcgtcttcac tcgaggagct tggcccattg catacgttgt atccatatca
      121 taatatgtac atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt
      181 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      241 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      361 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      421 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      481 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      541 taccatgcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
      601 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
      661 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgattgactg
      721 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
      781 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
      841 atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg
      901 gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat
      961 tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt
     1021 ggaactgacg agttcggaac acccggccgc aaccctggga gacgtcccag ggacttcggg
     1081 ggccgttttt gtggcccgac ctgagtccaa aaatcccgat cgttttggac tctttggtgc
     1141 acccccctta gaggagggat atgtggttct ggtaggagac gagaacctaa aacagttccc
     1201 gcctccgtct gaatttttgc tttcggtttg ggaccgaagc cgcgccgcgc gtcttgtctg
     1261 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatat
     1321 gggcccgggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     1381 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     1441 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     1501 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     1561 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     1621 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     1681 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     1741 cgcccccata tggccatatg agatcttata tggggcaccc ccgccccttg taaacttccc
     1801 tgaccctgac atgacaagag ttactaacag cccctctctc caagctcact tacaggctct
     1861 ctacttagtc cagcacgaag tctggagacc tctggcggca gcctaccaag aacaactgga
     1921 ccgaccggtg gtacctcacc cttaccgagt cggcgacaca gtgtgggtcc gccgacacca
     1981 gactaagaac ctagaacctc gctggaaagg accttacaca gtcctgctga ccacccccac
     2041 cgccctcaaa gtagacggca tcgcagcttg gatacacgcc gcccacgtga aggctgccga
     2101 ccccgggggt ggaccatcct ctagactgcc ggatcccagt gtggtggtac gggaattatg
     2161 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     2221 gacaaggaat tcggatctat ggaaccacag gttactctaa atgtgacttt taaaaatgaa
     2281 attcaaagct ttctggtttc tgatccagaa aatacaactt gggctgatat cgaagctatg
     2341 gtaaaagttt catttgatct gaatactatt caaataaaat acctggatga ggaaaatgaa
     2401 gaggtatcca tcaacagtca aggagaatat gaagaagcgc ttaagatggc agttaaacag
     2461 ggaaaccaac tgcagatgca agtccacgaa gggcaccatg tcgttgatga agccccaccc
     2521 ccagttgtag gagcaaaacg actagctgcc agggcaggga agaagccact tgcacattac
     2581 tcttcactgg tgagagtctt gggatcagac atgaagaccc cagaggatcc tgcagtgcag
     2641 tcgtttccac ttgttccatg tgacacagac cagcctcaag acaagccccc agactggttc
     2701 acaagctacc tggagacgtt cagagaacaa gtggttaacg aaacggttga gaagcttgaa
     2761 cagaaattac atgaaaagct tgtcctccag aacccatcct tgggttcttg tccctcagaa
     2821 gtctcaatgc ctacttcaga agaaacattg tttttgccag aaaaccagtt cagctggcat
     2881 attgcttgca acaactgcca aagaaggatt gttggtgtcc gctaccagtg tagcctatgc
     2941 ccatcctaca atatctgtga agattgtgaa gcagggccat atggccatga cactaaccac
     3001 gtcctgctga agttgcggag acctgttgtg ggctcctctg aaccgttctg tcactcaaag
     3061 tactctactc ctcgtcttcc tgctgctctg gaacaagtca ggctccagaa acaggttgat
     3121 aagaactttc ttaaagcaga aaagcaaagg ttgcgagctg agaagaaaca acgtaaagca
     3181 gaggtcaagg aacttaaaaa gcagcttaaa ctccatagga aaattcacct gtggaattca
     3241 atccatggac tccagagccc caagtctcct ttaggccgac ctgagagctt gctccagtct
     3301 aataccctga tgctcccttt gcagccctgt acctccgtta tgccaatgct cagtgcagca
     3361 tttgtggatg agaatttgcc tgatgggact caccttcagc caggaaccaa gtttatcaaa
     3421 cactggagga tgaaaaatac aggaaatgta aagtggagtg cagacacaaa gctcaagttc
     3481 atgtggggaa acctgacttt ggcttccaca gaaaagaagg atgttttggt tccctgcctc
     3541 aaggccggcc atgtgggagt tgtatctgtg gagttcattg ccccagcctt ggagggaacg
     3601 tatacttccc attggcgtct ttctcacaaa ggccagcaat ttgggcctcg ggtctggtgc
     3661 agtatcatag tagatccttt cccctccgaa gagagccctg ataacattga aaagggcatg
     3721 atcagctcaa gcaaaactga tgatctcacc tgccagcaag aggaaacttt tcttctggct
     3781 aaagaagaaa gacagcttgg tgaagtgact gagcagacag aagggacagc agcctgcatc
     3841 ccacagaagg caaaaaatgt tgccagtgag agggagctct acatcccatc tgtggatctt
     3901 ctgactgccc aggacctgct gtcctttgag ctgttggata taaacattgt tcaagagttg
     3961 gagagagtgc cccacaacac ccctgtggat gtgactccct gcatgtctcc tctgccacat
     4021 gacagtcctt taatagagaa gccaggcttg gggcagatag aggaagagaa tgaaggggca
     4081 ggatttaaag cacttcctga ttctatggtg tcagtaaaga ggaaggctga gaacattgct
     4141 tctgtggagg aagcagaaga agacctgagt gggacccagt ttgtgtgtga gacagtaatc
     4201 cgatccctta ccttggatgc tgccccagac cacaaccctc cttgcagaca gaagtccttg
     4261 cagatgacat ttgccttgcc tgaaggacca cttggaaatg agaaggagga gattatccat
     4321 atcgctgagg aagaagctgt catggaggag gaggaggatg aggaggatga ggaggaggag
     4381 gatgagctca aagatgaagt tcaaagtcag tcctctgctt cctcagagga ttacatcatc
     4441 atcctgcctg agtgctttga taccagccgc cccctggggg attctatgta cagctctgcg
     4501 ctctcacagc caggcctgga gcgaggtgct gaaggcaagc ctggggttga ggctgggcag
     4561 gaaccagctg aggctgggga aagactccct ggaggggaga accagccaca ggagcacagc
     4621 ataagtgaca tcctcacgac ctcacagact ctggaaacag tgcccctaat cccagaggta
     4681 gtggagcttc caccgtcact gcccaggagc tctccttgtg tacatcatca tggttcccca
     4741 ggagtggatt taccagttac cataccagaa gtttcttcag tccctgatca gatcagagga
     4801 gagcccagag gctcatcagg acttgtaaac agcagacaga agagctatga ccactcaagg
     4861 caccatcatg ggagcagcat tgctggagga ctggtgaagg gggctttgtc tgttgctgcc
     4921 tctgcataca aggccctgtt tgctgggcca ccagtcactg cacagccaat aatttgaaat
     4981 tcctgcaggc ctcgagggcc ggcgcgccgc ggccgctacg taaattccgc ccccccccct
     5041 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     5101 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     5161 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     5221 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     5281 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     5341 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     5401 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     5461 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     5521 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     5581 acacgatgat aatatggcca caaccatgac cgagtacaag cccacggtgc gcctcgccac
     5641 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5701 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5761 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5821 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5881 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5941 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     6001 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     6061 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     6121 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     6181 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     6241 gagcgcacga ccccatggct cgacgataaa ataaaagatt ttatttagtc tccagaaaaa
     6301 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     6361 caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga
     6421 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6481 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     6541 ttcctgcccc ggctcagggc caagaacaga tagtccccag atgcggtcca gccctcagca
     6601 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     6661 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     6721 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     6781 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     6841 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca
     6901 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat
     6961 agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7021 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7081 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7141 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7201 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7261 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7321 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7381 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7441 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7501 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7561 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7621 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7681 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7741 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7801 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7861 tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccggccgca aatcaatcta
     7921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7981 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     8581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgc
//
