LOCUS       sequence-405566-        9098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xFLAG-tagged NBR1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9098)
  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 9098)
  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
FEATURES             Location/Qualifiers
     source          1..9098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    957..1314
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1379..1795
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1725..1747
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1763..1779
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1805..2179
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2117..2133
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2212..2277
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5239..5805
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5255..5807
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5399..5416)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5626..5645
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5819..6418
                     /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(5819..5838)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6315..6335
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6642..7111
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7241..7258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7412..8000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7492..7511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8164..8955,8956..9024))
                     /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     8787..8806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9025..9098,1..31))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
       61 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
      121 ttcgtcttca ctcgaggagc ttggcccatt gcatacgttg tatccatatc ataatatgta
      181 catttatatt ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
      601 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      661 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      721 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
      781 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      841 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      901 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      961 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
     1021 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     1081 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
     1141 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt
     1201 agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     1261 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1321 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg
     1381 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     1441 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     1501 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1561 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1621 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1681 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1741 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     1801 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     1861 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     1921 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     1981 ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa
     2041 cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa
     2101 agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg
     2161 tggaccatcc tctagactgc cggatcccag tgtggtggta cgggaattat ggactacaaa
     2221 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaaggaa
     2281 ttcggatcta tggaaccaca ggttactcta aatgtgactt ttaaaaatga aattcaaagc
     2341 tttctggttt ctgatccaga aaatacaact tgggctgata tcgaagctat ggtaaaagtt
     2401 tcatttgatc tgaatactat tcaaataaaa tacctggatg aggaaaatga agaggtatcc
     2461 atcaacagtc aaggagaata tgaagaagcg cttaagatgg cagttaaaca gggaaaccaa
     2521 ctgcagatgc aagtccacga agggcaccat gtcgttgatg aagccccacc cccagttgta
     2581 ggagcaaaac gactagctgc cagggcaggg aagaagccac ttgcacatta ctcttcactg
     2641 gtgagagtct tgggatcaga catgaagacc ccagaggatc ctgcagtgca gtcgtttcca
     2701 cttgttccat gtgacacaga ccagcctcaa gacaagcccc cagactggtt cacaagctac
     2761 ctggagacgt tcagagaaca agtggttaac gaaacggttg agaagcttga acagaaatta
     2821 catgaaaagc ttgtcctcca gaacccatcc ttgggttctt gtccctcaga agtctcaatg
     2881 cctacttcag aagaaacatt gtttttgcca gaaaaccagt tcagctggca tattgcttgc
     2941 aacaactgcc aaagaaggat tgttggtgtc cgctaccagt gtagcctatg cccatcctac
     3001 aatatctgtg aagattgtga agcagggcca tatggccatg acactaacca cgtcctgctg
     3061 aagttgcgga gacctgttgt gggctcctct gaaccgttct gtcactcaaa gtactctact
     3121 cctcgtcttc ctgctgctct ggaacaagtc aggctccaga aacaggttga taagaacttt
     3181 cttaaagcag aaaagcaaag gttgcgagct gagaagaaac aacgtaaagc agaggtcaag
     3241 gaacttaaaa agcagcttaa actccatagg aaaattcacc tgtggaattc aatccatgga
     3301 ctccagagcc ccaagtctcc tttaggccga cctgagagct tgctccagtc taataccctg
     3361 atgctccctt tgcagccctg tacctccgtt atgccaatgc tcagtgcagc atttgtggat
     3421 gagaatttgc ctgatgggac tcaccttcag ccaggaacca agtttatcaa acactggagg
     3481 atgaaaaata caggaaatgt aaagtggagt gcagacacaa agctcaagtt catgtgggga
     3541 aacctgactt tggcttccac agaaaagaag gatgttttgg ttccctgcct caaggccggc
     3601 catgtgggag ttgtatctgt ggagttcatt gccccagcct tggagggaac gtatacttcc
     3661 cattggcgtc tttctcacaa aggccagcaa tttgggcctc gggtctggtg cagtatcata
     3721 gtagatcctt tcccctccga agagagccct gataacattg aaaagggcat gatcagctca
     3781 agcaaaactg atgatctcac ctgccagcaa gaggaaactt ttcttctggc taaagaagaa
     3841 agacagcttg gtgaagtgac tgagcagaca gaagggacag cagcctgcat cccacagaag
     3901 gcaaaaaatg ttgccagtga gagggagctc tacatcccat ctgtggatct tctgactgcc
     3961 caggacctgc tgtcctttga gctgttggat ataaacattg ttcaagagtt ggagagagtg
     4021 ccccacaaca cccctgtgga tgtgactccc tgcatgtctc ctctgccaca tgacagtcct
     4081 ttaatagaga agccaggctt ggggcagata gaggaagaga atgaaggggc aggatttaaa
     4141 gcacttcctg attctatggt gtcagtaaag aggaaggctg agaacattgc ttctgtggag
     4201 gaagcagaag aagacctgag tgggacccag tttgtgtgtg agacagtaat ccgatccctt
     4261 accttggatg ctgccccaga ccacaaccct ccttgcagac agaagtcctt gcagatgaca
     4321 tttgccttgc ctgaaggacc acttggaaat gagaaggagg agattatcca tatcgctgag
     4381 gaagaagctg tcatggagga ggaggaggat gaggaggatg aggaggagga ggatgagctc
     4441 aaagatgaag ttcaaagtca gtcctctgct tcctcagagg attacatcat catcctgcct
     4501 gagtgctttg ataccagccg ccccctgggg gattctatgt acagctctgc gctctcacag
     4561 ccaggcctgg agcgaggtgc tgaaggcaag cctggggttg aggctgggca ggaaccagct
     4621 gaggctgggg aaagactccc tggaggggag aaccagccac aggagcacag cataagtgac
     4681 atcctcacga cctcacagac tctggaaaca gtgcccctaa tcccagaggt agtggagctt
     4741 ccaccgtcac tgcccaggag ctctccttgt gtacatcatc atggttcccc aggagtggat
     4801 ttaccagtta ccataccaga agtttcttca gtccctgatc agatcagagg agagcccaga
     4861 ggctcatcag gacttgtaaa cagcagacag aagagctatg accactcaag gcaccatcat
     4921 gggagcagca ttgctggagg actggtgaag ggggctttgt ctgttgctgc ctctgcatac
     4981 aaggccctgt ttgctgggcc accagtcact gcacagccaa taatttctga agatcagaca
     5041 gcagccctga tggcccatct ctttgaaatg ggattctgtg acaggcagct gaacctacgg
     5101 ctgctgaaga aacacaatta caatatcctg caggttgtga cagaacttct tcagttaaac
     5161 aacaacgact ggtacagcca acgctattga aattcctgca ggcctcgagg gccggcgcgc
     5221 cgcggccgct acgtaaattc cgcccccccc cctaacgtta ctggccgaag ccgcttggaa
     5281 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     5341 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     5401 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     5461 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     5521 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     5581 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     5641 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     5701 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     5761 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     5821 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     5881 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     5941 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     6001 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     6061 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     6121 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     6181 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     6241 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     6301 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6361 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     6421 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg gctcgacgat
     6481 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     6541 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     6601 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     6661 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     6721 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     6781 agatagtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     6841 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     6901 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     6961 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     7021 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     7081 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
     7141 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     7201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8101 agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8161 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8221 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8281 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8341 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8461 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8581 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8761 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     8821 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9001 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9061 gggttattgt ctcatgagcg gatacatatt tgaatgta
//
