LOCUS       sequence-252695-        7453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of histone H3.3K4M-FLAG/HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7453)
  AUTHORS   Jang Y, Broun A, Wang C, Park YK, Zhuang L, Lee JE, Froimchuk E, Liu
            C, Ge K
  TITLE     H3.3K4M destabilizes enhancer H3K4 methyltransferases MLL3/MLL4 and 
            impairs adipose tissue development.
  JOURNAL   Nucleic Acids Res. 2019 Jan 25;47(2):607-620. doi: 
            10.1093/nar/gky982.
  PUBMED    30335158
REFERENCE   2  (bases 1 to 7453)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2019-01-25- 25"; volume: "47"; issue: 
            "2"; pages: "607-620"
COMMENT     Sequence Label: pMSCVhyg-H3.3K4M-FLAG/HA
FEATURES             Location/Qualifiers
     source          1..7453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1851..1874
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1890..1916
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1890..1916
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        1946..2445
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1981..2004)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2378..2397
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2463..3488
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2463..3488
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             3846..4360
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4522..4538
                     /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(4546..4552,4553..4570,4571..4576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4591..4612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4729..4746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4900..5488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4980..4999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5659..6450,6451..6519))
                     /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     6282..6301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6520..6624)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6692..6710
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6748..6770)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6870..6889
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7083..7105
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7233..7252
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7452..7453,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga gtgaaccatg gcccgaacca
     1441 tgcagactgc tcgtaagtcc accggtggga aagccccccg caaacagctg gccacgaaag
     1501 ccgccaggaa aagcgctccc tctaccggcg gggtgaagaa gcctcatcgc tacaggcccg
     1561 ggaccgtggc gcttcgagag attcgtcgtt atcagaagtc gaccgagctg ctcatccgga
     1621 agctgccctt ccagaggttg gtgagggaga tcgcgcagga tttcaaaacc gacctgaggt
     1681 ttcagagcgc agccatcggt gcgctgcagg aggctagcga agcgtacctg gtgggtctgt
     1741 tcgaagatac caacctgtgt gccatccacg ctaagagagt caccatcatg cccaaagaca
     1801 tccagttggc tcgccggata cggggagaga gagctgcggc cgctggagga gactacaagg
     1861 acgacgatga caagtcggcc gctggaggat acccctacga cgtgcccgac tacgcctagg
     1921 aattcgttaa cgaacgaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     1981 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     2041 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     2101 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     2161 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     2221 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     2281 ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg
     2341 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     2401 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg catcccgcca
     2461 ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     2521 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     2581 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     2641 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     2701 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     2761 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     2821 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     2881 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     2941 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     3001 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     3061 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     3121 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     3181 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     3241 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     3301 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     3361 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     3421 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     3481 aggaatagag tagatgccga ccgaacaaga gctgatttcg agaacgcctc agccagcaac
     3541 tcgcgcgagc ctagcaaggc aaatgcgaga gaacggcctt acgcttggtg gcacagttct
     3601 cgtccacagt tcgctaagct cgctcggctg ggtcgcggga gggccggtcg cagtgattca
     3661 ggcccttctg gattgtgttg gtccccaggg cacgattgtc atgcccacgc actcgggtga
     3721 tctgactgat cccgcagatt ggagatcgcc gcccgtgcct gccgattggg tgcagatccg
     3781 tcgacctgca gccaagctta tcgataaaat aaaagatttt atttagtctc cagaaaaagg
     3841 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     3901 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     3961 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     4021 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     4081 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     4141 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     4201 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
     4261 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     4321 cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt
     4381 tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca
     4441 ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa
     4501 gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4561 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4621 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5341 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6661 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6721 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6781 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6841 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6901 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6961 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     7021 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     7081 ttcccagtca cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg
     7141 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     7201 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     7261 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc
     7321 caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac
     7381 cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag
     7441 aaaaaggggg gaa
//
