LOCUS       sequence-438535-        8687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NSD3 (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8687)
  AUTHORS   Landau LM, Chaudhary N, Tien YC, Rogozinska M, Joshi S, Yao C, 
            Crowley J, Hullahalli K, Campbell IW, Waldor MK, Haigis M, Kagan JC
  TITLE     pLxIS-containing domains are biochemically flexible regulators of 
            interferons and metabolism.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2436-2454.e10. doi: 
            10.1016/j.molcel.2024.05.030. Epub 2024 Jun 25.
  PUBMED    38925114
REFERENCE   2  (bases 1 to 8687)
  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: 
            "10.1016/j.molcel.2024.05.030"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2436-2454.e10"
COMMENT     Sequence Label: pMSCV-FKBP-NSD3
FEATURES             Location/Qualifiers
     source          1..8687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16..81
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2310..2876
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2326..2878
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2470..2487)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2697..2716
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2890..2892,2893..2895,2896..3609)
                     /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(2935..2956)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3196..3215)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3543..3564
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3671..4185
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4347..4363
                     /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(4371..4377,4378..4395,4396..4401))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4416..4437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4554..4571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4725..5313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4805..4824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5484..6275,6276..6344))
                     /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     6107..6126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6345..6449)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6517..6535
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6573..6595)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6695..6714
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6908..6930
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7058..7077
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7277..7793
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7857..8198
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8265..8681
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8611..8633
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8649..8665
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaggatcac gacggtgatt acaaggacca cgacatcgac
       61 tacaaggacg acgacgacaa gggcgttcaa gtagaaacaa tatctccagg agacgggcgt
      121 acatttccta agcgtggcca aacttgcgta gtccattata ccggcatgct ggaggacggt
      181 aagaaggtcg atagctctag agaccgtaac aaacctttca aatttatgct ggggaaacag
      241 gaggttatcc gtggttggga ggaaggcgtg gcccaaatgt ccgttgggca gcgtgctaag
      301 ttaaccatct cgccagatta cgcatatggg gctacaggcc atcctgggat cattcccccg
      361 catgcgacac ttgtgtttga tgtagagctg ttgaagttgg agacgcgggg agttcaagtc
      421 gaaacgattt caccaggtga tggacgcacc tttcctaaac ggggacaaac gtgcgtcgtg
      481 cattacaccg gtatgctgga agacggcaag aaagtcgatt caagccgcga tagaaataaa
      541 cccttcaaat tcatgcttgg aaaacaggaa gtaatacgcg gatgggaaga gggtgtggcg
      601 caaatgtctg tcgggcaaag agctaaactg acaataagtc ctgattacgc gtacggtgcg
      661 actgggcacc ctggtatcat tccgccgcac gcgacacttg tcttcgacgt cgagttactt
      721 aagcttgaga cgcggggggt gcaggtagag actatttcgc ccggtgacgg aagaactttc
      781 ccaaagcggg gacagacctg tgttgtccac tacactggga tgttggagga tggtaagaaa
      841 gttgactcaa gtcgtgatcg caacaaaccc tttaagttta tgcttgggaa gcaggaagtc
      901 atcagaggtt gggaagaagg cgtagctcag atgtcagtgg gacaacgggc caaattaacc
      961 atcagtccag actacgccta cggagccacc ggtcacccag ggatcatacc acctcacgca
     1021 accttagtgt tcgatgtgga acttttaaag ttagagacgc gcggtgtaca agtggaaaca
     1081 atcagtccag gggatgggag aacattcccc aaaagaggcc agacgtgtgt cgtgcactat
     1141 acggggatgt tagaagatgg gaaaaaggtc gattcgtcac gcgaccgcaa caaaccgttt
     1201 aaattcatgt tgggaaagca ggaggtgatt cgcggctggg aggaaggtgt ggcacagatg
     1261 agcgtgggcc agagagcaaa actgactatc tctcctgatt atgcctacgg cgcgacgggc
     1321 caccctggca taattccacc ccacgctacc ctggtattcg atgtcgagct tctgaagctt
     1381 gagggtgctg gggctggggc tgataagcag ccagaagagg cttcgtccca agcaaagaag
     1441 aatgttacct ctaagacgga agtcaagaaa ccccgaagac caaggagcgt gctgaacagt
     1501 cagccagaac agaccaatgc tggggaggtg gcctcctcac aatcaagtac tgaccttcga
     1561 aggcagagcc agaggcggca tactagcttg gaagaggaag agccacctcc tgttaaaatc
     1621 gcctggaaaa cagcagccgc aaggaagtcc ttaccagcct ccatcacaat gcacaaaggg
     1681 agcctagatt tgcagaagtg taatatgtct ccagttgtga aaattgaaca agtgtttgct
     1741 ctccagaatg caacaggaga tgggaagttt attgatcagt ttgtttattc aacgaagggt
     1801 attggtaaca aaacagaaat aagtgtcagg gggcaagaca ggcttataat ctcttcacca
     1861 agtcagagaa gtgaaaagcc ggctcagagc gcgtcatctc cggaggcaac atctggttct
     1921 gcaggcccag tagagaagaa gcagcagaga agatccatca ggactcgatc tgagtcagag
     1981 aagtccgccg aggttgtgcc aaagaagaag atcaaaaagg agcaggttga gacagctccc
     2041 caggcctccc tgaagaccgg gttacagaaa ggtgccagcg agatttcaga ttcctgtaag
     2101 cccctaaaga aaaggagtcg cgcctcaacg gatgtagaga cggctagttg cacgtacaga
     2161 gacacatctg actccgattc tagaggactg agtgacgggc aggtaggctt tggcaaacaa
     2221 gtagatagcc cgtcagctac tgctgacgca gatgcttgat aagcggccgc gtttaaacgt
     2281 cgacggtatc gataagcttg atatcgaatt ccgccccccc ccctaacgtt actggccgaa
     2341 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     2401 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     2461 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     2521 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     2581 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     2641 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     2701 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     2761 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     2821 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     2881 gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2941 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat
     3001 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3061 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3121 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3181 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3241 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3301 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3361 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3421 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3481 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3541 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3601 tacaagtaat gaattaatta agaattcgat aaaataaaag attttattta gtctccagaa
     3661 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     3721 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     3781 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3841 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     3901 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     3961 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     4021 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     4081 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     4141 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     4201 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     4261 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     4321 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga
     4381 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4441 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4501 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4561 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4621 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4681 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4741 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4801 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4861 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4921 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4981 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5041 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5101 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5161 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5221 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5281 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5341 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5401 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5461 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5521 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5581 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5641 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5701 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5761 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5821 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5881 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5941 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6001 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6061 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6121 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6181 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6241 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6301 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6361 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6421 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6481 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6541 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6601 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6661 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6721 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6781 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6841 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6901 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga
     6961 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     7021 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     7081 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat
     7141 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat
     7201 atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc
     7261 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     7321 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     7381 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     7441 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     7501 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     7561 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     7621 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     7681 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc
     7741 cttgggaggg tctcctcaga ttgattgact gcccacctcg ggggtctttc atttggaggt
     7801 tccaccgaga tttggagacc cctgcccagg gaccaccgac ccccccgccg ggaggtaagc
     7861 tggccagcgg tcgtttcgtg tctgtctctg tctttgtgcg tgtttgtgcc ggcatctaat
     7921 gtttgcgcct gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg
     7981 gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg
     8041 gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat
     8101 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
     8161 tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt
     8221 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca
     8281 ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt
     8341 cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta
     8401 acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca
     8461 aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg
     8521 aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc
     8581 ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc
     8641 gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggaatt
//
