LOCUS       sequence-393709-        6927 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of NINJ1 gene in mammalian cells by retroviral 
            transduction .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6927)
  AUTHORS   Borges JP, Saetra RSR, Volchuk A, Bugge M, Devant P, Sporsheim B, 
            Kilburn BR, Evavold CL, Kagan JC, Goldenberg NM, Flo TH, Steinberg 
            BE
  TITLE     Glycine inhibits NINJ1 membrane clustering to suppress plasma 
            membrane rupture in cell death.
  JOURNAL   Elife. 2022 Dec 5;11:e78609. doi: 10.7554/eLife.78609.
  PUBMED    36468682
REFERENCE   2  (bases 1 to 6927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2022-12-5"; volume: "11:e78609. doi"; pages: " 
            10.7554/eLife.78609"
COMMENT     Sequence Label: pMSCV-NINJ1
FEATURES             Location/Qualifiers
     source          1..6927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1440..1449
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1449..1472
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1960..2526
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1976..2528
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2120..2137)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2347..2366
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2540..2542,2543..2545,2546..3259)
                     /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(2585..2606)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2846..2865)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3193..3214
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3321..3835
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3997..4013
                     /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(4021..4027,4028..4045,4046..4051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4066..4087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4204..4221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4375..4963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4455..4474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5134..5925,5926..5994))
                     /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     5757..5776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5995..6099)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6167..6185
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6223..6245)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6345..6364
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6558..6580
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6708..6727
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6927,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctctcg agatcgatgc ggccgcgccg
     1441 ccaccatgga ctacaaagac gatgacgaca aggagtctgg caccgaggaa tatgagctga
     1501 acggtgattt gcgccctggc tctcctggtt ctcctgatgc cctcccacca aggtgggggc
     1561 ttcggaatcg ccctatcaat gtaaatcact acgcaaataa gaaatcagca gcagagtcta
     1621 tgctggacat tgcactcttg atggctaacg cttctcagtt gaaggctgtg gtagagcaag
     1681 gaaacgattt tgcatttttt gtccccttgg tggtattgat ttcaataagc ctggtacttc
     1741 agattggagt cggggtactc ctgatcttct tggtgaaata tgacctcaac aatccagcca
     1801 aacacgcaaa gctggatttt cttaataatc tcgcaaccgg actggtattt attatcgtag
     1861 tcgttaacat ttttatcacc gcatttgggg tccagaagcc agtgatggat gtcgccccaa
     1921 gacaataggt cgacggtatc gataagcttg atatcgaatt ccgccccccc ccctaacgtt
     1981 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     2041 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     2101 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     2161 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     2221 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2281 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2341 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2401 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     2461 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     2521 tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2581 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg
     2641 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2701 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2761 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2821 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2881 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2941 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3001 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     3061 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3121 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     3181 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3241 ggacgagctg tacaagtaat gaattaatta agaattcgat aaaataaaag attttattta
     3301 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     3361 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     3421 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     3481 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     3541 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     3601 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     3661 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     3721 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     3781 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta
     3841 acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc
     3901 tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta
     3961 tggacagcgc agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca
     4021 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4081 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4141 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5401 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6121 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6181 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6241 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6301 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6361 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6421 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6481 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6541 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca
     6601 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     6661 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     6721 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     6781 tagaggcgat tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg
     6841 actcaacaat atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt
     6901 ttatttagtc tccagaaaaa gggggga
//
