LOCUS       sequence-405564-        6200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xFLAG tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6200)
  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 6200)
  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
FEATURES             Location/Qualifiers
     source          1..6200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..71
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        173..552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    911..1268
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1333..1749
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1679..1701
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1717..1733
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1759..2133
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2071..2087
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2166..2231
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2295..2861
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2311..2863
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2455..2472)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2682..2701
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2875..3474
                     /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(2875..2894)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3371..3391
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3698..4167
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4297..4314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4468..5056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4548..4567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5220..6011,6012..6080))
                     /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     5843..5862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6081..6185)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
       61 tatcacgagg ccctttcgtc ttcactcgag gagcttggcc cattgcatac gttgtatcca
      121 tatcataata tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga
      181 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      241 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      301 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      361 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      421 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      481 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      541 gctattacca tgctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
      601 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
      661 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgatt
      721 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
      781 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
      841 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
      901 accgggaggt aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga
      961 ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc
     1021 gtggtggaac tgacgagttc ggaacacccg gccgcaaccc tgggagacgt cccagggact
     1081 tcgggggccg tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt tggactcttt
     1141 ggtgcacccc ccttagagga gggatatgtg gttctggtag gagacgagaa cctaaaacag
     1201 ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt
     1261 gtctgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa
     1321 aatatgggcc cgggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     1381 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     1441 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     1501 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     1561 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     1621 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     1681 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     1741 ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc ccttgtaaac
     1801 ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc tcacttacag
     1861 gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta ccaagaacaa
     1921 ctggaccgac cggtggtacc tcacccttac cgagtcggcg acacagtgtg ggtccgccga
     1981 caccagacta agaacctaga acctcgctgg aaaggacctt acacagtcct gctgaccacc
     2041 cccaccgccc tcaaagtaga cggcatcgca gcttggatac acgccgccca cgtgaaggct
     2101 gccgaccccg ggggtggacc atcctctaga ctgccggatc ccagtgtggt ggtacgggaa
     2161 ttatggacta caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg
     2221 acgatgacaa ggaattcgga tcttgaaatt cctgcaggcc tcgagggccg gcgcgccgcg
     2281 gccgctacgt aaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     2341 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     2401 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     2461 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     2521 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     2581 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     2641 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     2701 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     2761 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     2821 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgacc
     2881 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc
     2941 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac
     3001 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3061 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3121 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3181 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     3241 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     3301 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     3361 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     3421 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     3481 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc gacgataaaa
     3541 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     3601 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     3661 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     3721 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     3781 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     3841 agtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     3901 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     3961 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     4021 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     4081 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4141 tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt
     4201 tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg
     4261 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5161 ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5221 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5281 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5341 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5401 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5461 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5521 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5581 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5641 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5701 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5761 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5821 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5881 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5941 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6001 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6061 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6121 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6181 ccgcgcacat ttccccgaaa
//
