LOCUS       sequence-379078-        8867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of HaloTag7-NCOA4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8867)
  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 8867)
  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-HaloTag7-NCOA4
FEATURES             Location/Qualifiers
     source          1..8867
                     /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             1994..2881
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     misc_feature    4778..5344
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4794..5346
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4938..4955)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5165..5184
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5364..5963
                     /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(5364..5383)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5860..5880
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6193..6662
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6792..6809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6963..7551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7043..7062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7715..8506,8507..8575))
                     /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     8338..8357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8576..8680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8748..8766
                     /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 accatggaaa tcggtactgg ctttccattc gacccccatt atgtggaagt
     2041 cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt
     2101 cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc
     2161 gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct
     2221 gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct
     2281 ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa
     2341 gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac
     2401 ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt
     2461 cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt
     2521 cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt
     2581 tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa
     2641 catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct
     2701 gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa
     2761 aagcctgcct aactgcaagg ctgtggacat cggcccaggt ctgaatctgc tgcaagaaga
     2821 caacccggac ctgatcggca gcgagatcgc gcgctggctg tctactctgg agatttccgg
     2881 tggttcaggc tcgggatccg ggaattcgaa taccttccaa gaccagagtg gcagctccag
     2941 taatagagaa ccccttttga ggtgtagtga tgcacggagg gacttggagc ttgctattgg
     3001 tggagttctc cgggctgaac agcaaattaa agataacttg cgagaggtca aagctcagat
     3061 tcacagttgc ataagccgtc acctggaatg tcttagaagc cgtgaggtat ggctgtatga
     3121 acaggtggac cttatttatc agcttaaaga ggagacactt caacagcagg ctcagcagct
     3181 ctactcgtta ttgggccagt tcaattgtct tactcatcaa ctggagtgta cccaaaacaa
     3241 agatctagcc aatcaagtct ctgtgtgcct ggagagactg ggcagtttga cccttaagcc
     3301 tgaagattca actgtcctgc tctttgaagc tgacacaatt actctgcgcc agaccatcac
     3361 cacatttggg tctctcaaaa ccattcaaat tcctgagcac ttgatggctc atgctagttc
     3421 agcaaatatt gggcccttcc tggagaagag aggctgtatc tccatgccag agcagaagtc
     3481 agcatccggt attgtagctg tccctttcag cgaatggctc cttggaagca aacctgccag
     3541 tggttatcaa gctccttaca tacccagcac cgacccccag gactggctta cccaaaagca
     3601 gaccttggag aacagtcaga cttcttccag agcctgcaat ttcttcaata atgtcggggg
     3661 aaacctaaag ggcttagaaa actggctcct caagagtgaa aaatcaagtt atcaaaagtg
     3721 taacagccat tccactacta gttctttctc cattgaaatg gaaaaggttg gagatcaaga
     3781 gcttcctgat caagatgaga tggacctatc agattggcta gtgactcccc aggaatccca
     3841 taagctgcgg aagcctgaga atggcagtcg tgaaacttct gaaaaattca aattattgtt
     3901 ccagtcctat aatgtgaatg attggcttgt caagactgac tcctgtacca actgtcaggg
     3961 aaaccagccc aaaggtgtgg agattgaaaa cctgggcaat ctgaagtgcc tgaatgacca
     4021 cttggaggcc aagaaaccat tgtccacccc cagcatggtt acagaggatt ggcttgtcca
     4081 gaaccatcag gacccatgta aggtagagga ggtgtgcaga gccaatgagc cctgcacaag
     4141 ctttgcagag tgtgtgtgtg atgagaattg tgagaaggag gctctgtata agtggcttct
     4201 gaagaaagaa ggaaaggata aaaatgggat gcctgtggaa cccaaacctg agcctgagaa
     4261 gcataaagat tccctgaata tgtggctctg tcctagaaaa gaagtaatag aacaaactaa
     4321 agcaccaaag gcaatgactc cttctagaat tgctgattcc ttccaagtca taaagaacag
     4381 ccccttgtcg gagtggctta tcaggccccc atacaaagaa ggaagtccca aggaagtgcc
     4441 tggtactgaa gacagagctg gcaaacagaa gtttaaaagc cccatgaata cttcctggtg
     4501 ttcctttaac acagctgact gggtcctgcc aggaaagaag atgggcaacc tcagccagtt
     4561 atcttctgga gaagacaagt ggctgcttcg aaagaaggcc caggaagtat tacttaattc
     4621 acctctacag gaggaacata acttcccccc agaccattat ggcctccctg cagtttgtga
     4681 tctctttgcc tgtatgcagc ttaaagttga taaagagaag tggttatatc gaactcctct
     4741 acagatgtag ctcgagtaaa gcggccgcta cgtaaattcc gccccccccc ctaacgttac
     4801 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     4861 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     4921 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     4981 agcagttcct ctggaagttt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     5041 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     5101 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     5161 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     5221 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     5281 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     5341 ataatatggc cacaaccgtc gacatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     5401 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     5461 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     5521 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     5581 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     5641 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     5701 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     5761 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5821 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     5881 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5941 tgacccgcaa gcccggtgcc tgagtcgacc gcccgcccca cgacccgcag cgcccgaccg
     6001 aaaggagcgc acgaccccat ggctcgacga taaaataaaa gattttattt agtctccaga
     6061 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     6121 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     6181 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     6241 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     6301 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     6361 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     6421 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     6481 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     6541 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     6601 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     6661 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     6721 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6781 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6841 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6901 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7381 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc cgcaaatcaa
     7681 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7801 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7921 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8161 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8221 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     8341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8401 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8581 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8641 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8701 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     8761 cgaggccctt tcgtcttcac tataggagct tggcccattg catacgttgt atccatatca
     8821 taatatgtac atttatattg gctcatgtcc aacattaccg ccatgtt
//
