LOCUS       sequence-411195-        6914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MSCV vector expressing K45Q mouse NINJ1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6914)
  AUTHORS   Ramos S, Hartenian E, Santos JC, Walch P, Broz P
  TITLE     NINJ1 induces plasma membrane rupture and release of 
            damage-associated molecular pattern molecules during ferroptosis.
  JOURNAL   EMBO J. 2024 Apr;43(7):1164-1186. doi: 10.1038/s44318-024-00055-y. 
            Epub 2024 Feb 23.
  PUBMED    38396301
REFERENCE   2  (bases 1 to 6914)
  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"; doi: 
            "10.1038/s44318-024-00055-y"; journalName: "EMBO J"; date: 
            "2024-04"; volume: "43"; issue: "7"; pages: "1164-1186"
COMMENT     Sequence Label: MSCV-mNINJ1-K45Q-GFP
FEATURES             Location/Qualifiers
     source          1..6914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     96..112
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      160..169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    671..1237
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    687..1239
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(831..848)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1058..1077
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1251..1253,1254..1256,1257..1970)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1296..1317)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1557..1576)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1904..1925
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2032..2546
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2708..2724
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2732..2738,2739..2756,2757..2762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2777..2798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2915..2932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3086..3674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3166..3185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3845..4636,4637..4705))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4468..4487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4706..4810)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4878..4896
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4934..4956)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5056..5075
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5269..5291
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5419..5438
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5638..6154
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6218..6559
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(6626..6914,1..128)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct ctcgagatcg atgcggcccg ccaccatgga gtcgggcact
      181 gaggagtatg agctcaacgg cgacctgcgc ccgggctccc ccggttcccc cgacgccttg
      241 ccaccccgct ggggtttgcg gaaccggccc atcaatgtaa accattatgc caacaagcag
      301 agcgctgcgg agagcatgct ggacatcgcg ctgctcatgg ccaacgcgtc gcagctgaag
      361 gccgtggtgg agcagggcaa tgatttcgcc ttcttcgtgc cccttgtggt cctcatctct
      421 atctccctcg tgctgcagat aggagtgggc gtgctgctca tcttcctggt caagtatgac
      481 ctcaacaacc cggccaagca cgccaagctg gactttctta acaacctggc cacgggactg
      541 gttttcatca tcgtcgtggt caacatcttc attacggcct tcggggtcca gaagcctgta
      601 atggacgtgg cgccccggca gtaagggccg cgtttaaacg tcgacggtat cgataagctt
      661 gatatcgaat tccgcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
      721 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
      781 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
      841 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
      901 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
      961 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     1021 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     1081 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     1141 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     1201 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca
     1261 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1321 acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga
     1381 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1441 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1501 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1561 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1621 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1681 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1741 cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1801 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1861 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1921 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa tgaattaatt
     1981 aagaattcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     2041 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
     2101 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
     2161 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     2221 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     2281 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     2341 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     2401 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct
     2461 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     2521 actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt
     2581 ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca
     2641 catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc tggggagaat
     2701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2821 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2881 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2941 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3001 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3061 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3121 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3421 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3481 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3541 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3601 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3661 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3721 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3781 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3841 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3901 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3961 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4021 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4081 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4141 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4201 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4261 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4321 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4381 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4441 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4501 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4561 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4621 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4681 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4801 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4861 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4921 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4981 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5041 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     5101 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     5161 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     5221 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     5281 gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     5341 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     5401 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     5461 gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga
     5521 caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat
     5581 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     5641 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     5701 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     5761 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     5821 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     5881 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     5941 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     6001 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgta ttcccaataa
     6061 agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg gtctcctcag
     6121 attgattgac tgcccacctc gggggtcttt catttggagg ttccaccgag atttggagac
     6181 ccctgcccag ggaccaccga cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt
     6241 gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta
     6301 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
     6361 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
     6421 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
     6481 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
     6541 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
     6601 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     6661 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     6721 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     6781 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     6841 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     6901 ccctccctgg gtca
//
