LOCUS       sequence-379073-        9095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of mGFP-TAX1BP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9095)
  AUTHORS   Ohshima T, Yamamoto H, Sakamaki Y, Saito C, Mizushima N
  TITLE     NCOA4 drives ferritin phase separation to facilitate 
            macroferritinophagy and microferritinophagy.
  JOURNAL   J Cell Biol. 2022 Oct 3;221(10). pii: 213442. doi: 
            10.1083/jcb.202203102. Epub 2022 Sep 6.
  PUBMED    36066504
REFERENCE   2  (bases 1 to 9095)
  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: "J Cell 
            Biol."; date: "2022-10-3"; volume: "221(10). pii: 213442. doi"; 
            pages: " 10.1083/jcb.202203102. Epub 2022 Sep 6"
COMMENT     Sequence Label: pMRX-IP-mGFP-TAX1BP1
FEATURES             Location/Qualifiers
     source          1..9095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1507..1529
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1545..1561
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1899..1915
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1988..1997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1994..1996,1997..1999,2000..2710)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2039..2060)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2300..2319)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2647..2668
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5006..5572
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5022..5574
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5166..5183)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5393..5412
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5592..6191
                     /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(5592..5611)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6088..6108
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6421..6890
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7020..7037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7191..7779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7271..7290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7943..8734,8735..8803))
                     /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     8566..8585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8804..8908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8976..8994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
      601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
      661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
      721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
      841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
      901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
      961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat
     1981 aaagcttgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2041 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2101 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2161 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2221 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2281 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2341 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2401 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2461 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2521 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2581 gcccgacaac cactacctga gcacccagtc caaactgagc aaagacccca acgagaagcg
     2641 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2701 gctgtacaag ggttcaggct cgggatccgg gaattcgaca tcctttcaag aagtcccatt
     2761 gcagacttcc aactttgccc atgtcatctt tcaaaatgtg gccaagagtt accttcctaa
     2821 tgcacacctg gaatgtcatt acaccttaac tccatatatt catccacatc caaaagattg
     2881 ggttggtata ttcaaggttg gatggagtac tgctcgtgat tattacacgt ttttatggtc
     2941 ccctatgcct gaacattatg tggaaggatc aacagtcaat tgtgtactag cattccaagg
     3001 atattacctt ccaaatgatg atggagaatt ttatcagttc tgttacgtta cccataaggg
     3061 tgaaattcgt ggagcaagta cacctttcca gtttcgagct tcttctccag ttgaagagct
     3121 gcttactatg gaagatgaag gaaattctga catgttagtg gtgaccacaa aagcaggcct
     3181 tcttgagttg aaaattgaga aaaccatgaa agaaaaagaa gaactgttaa agttaattgc
     3241 cgttctggaa aaagaaacag cacaacttcg agaacaagtt gggagaatgg aaagagaact
     3301 taaccatgag aaagaaagat gtgaccaact gcaagcagaa caaaagggtc ttactgaagt
     3361 aacacaaagc ttaaaaatgg aaaatgaaga gtttaagaag aggttcagtg atgctacatc
     3421 caaagcccat cagcttgagg aagatattgt gtcagtaaca cataaagcaa ttgaaaaaga
     3481 aaccgaatta gacagtttaa aggacaaact caagaaggca caacatgaaa gagaacaact
     3541 tgaatgtcag ttgaagacag agaaggatga aaaggaactt tataaggtac atttgaagaa
     3601 tacagaaata gaaaatacca agcttatgtc agaggtccag actttaaaaa atttagatgg
     3661 gaacaaagaa agcgtgatta ctcatttcaa agaagagatt ggcaggctgc agttatgttt
     3721 ggctgaaaag gaaaatctgc aaagaacttt cctgcttaca acctcaagta aagaagatac
     3781 ttgtttttta aaggagcaac ttcgtaaagc agaggaacag gttcaggcaa ctcggcaaga
     3841 agttgtcttt ctggctaaag aactcagtga tgctgtcaac gtacgagaca gaacgatggc
     3901 agacctgcat actgcacgct tggaaaacga gaaagtgaaa aagcagttag ctgatgcagt
     3961 ggcagaactt aaactaaatg ctatgaaaaa agatcaggac aagactgata cactggaaca
     4021 cgaactaaga agagaagttg aagatctgaa actccgtctt cagatggctg cagaccatta
     4081 taaagaaaaa tttaaggaat gccaaaggct ccaaaaacaa ataaacaaac tttcagatca
     4141 atcagctaat aataataatg tcttcacaaa gaaaacgggg aatcagcaga aagtgaatga
     4201 tgcttcagta aacacagacc cagccacttc tgcctctact gtagatgtaa agccatcacc
     4261 ttctgcagca gaggcagatt ttgacatagt aacaaagggg caagtctgtg aaatgaccaa
     4321 agaaattgct gacaaaacag aaaagtataa taaatgtaaa caactcttgc aggatgagaa
     4381 agcaaaatgc aataaatatg ctgatgaact tgcaaaaatg gagctgaaat ggaaagaaca
     4441 agtgaaaatt gctgaaaatg taaaacttga actagctgaa gtacaggaca attataaaga
     4501 acttaaaagg agtctagaaa atccagcaga aaggaaaatg gaagatggag cagatggtgc
     4561 tttttaccca gatgaaatac aaaggccacc tgtcagagtc ccctcttggg gactggaaga
     4621 caatgttgtc tgcagccagc ctgctcgaaa ctttagtcgg cctgatggct tagaggactc
     4681 tgaggatagc aaagaagatg agaatgtgcc tactgctcct gatcctccaa gtcaacattt
     4741 acgtgggcat gggacaggct tttgctttga ttccagcttt gatgttcaca agaagtgtcc
     4801 cctctgtgag ttaatgtttc ctcctaacta tgatcagagc aaatttgaag aacatgttga
     4861 aagtcactgg aaggtgtgcc cgatgtgcag cgagcagttc cctcctgact atgaccagca
     4921 ggtgtttgaa aggcatgtgc agacccattt tgatcagaat gttctaaatt ttgactagct
     4981 cgagtaaagc ggccgctacg taaattccgc ccccccccct aacgttactg gccgaagccg
     5041 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     5101 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     5161 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     5221 ggaagtttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     5281 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     5341 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     5401 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     5461 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     5521 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     5581 caaccgtcga catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5641 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5701 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5761 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5821 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5881 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5941 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     6001 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     6061 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     6121 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     6181 ccggtgcctg agtcgaccgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     6241 gaccccatgg ctcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     6301 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     6361 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     6421 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     6481 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     6541 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     6601 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     6661 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     6721 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     6781 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     6841 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     6901 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     6961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7621 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7861 tcaaaaagga tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat
     7921 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7981 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8041 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8101 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8161 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8221 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8281 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8341 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8401 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8461 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8521 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     8581 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8641 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8701 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8761 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8881 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8941 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     9001 gtcttcacta taggagcttg gcccattgca tacgttgtat ccatatcata atatgtacat
     9061 ttatattggc tcatgtccaa cattaccgcc atgtt
//
