LOCUS       sequence-170477-        8695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of human SMYD3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8695)
  AUTHORS   Mazur PK, Reynoird N, Khatri P, Jansen PW, Wilkinson AW, Liu S, 
            Barbash O, Van Aller GS, Huddleston M, Dhanak D, Tummino PJ, Kruger 
            RG, Garcia BA, Butte AJ, Vermeulen M, Sage J, Gozani O
  TITLE     SMYD3 links lysine methylation of MAP3K2 to Ras-driven cancer.
  JOURNAL   Nature. 2014 Jun 12;510(7504):283-7. doi: 10.1038/nature13320. Epub 
            2014 May 21.
  PUBMED    24847881
REFERENCE   2  (bases 1 to 8695)
  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/nature13320"; journalName: "Nature"; date: "2014-06-12- 
            12"; volume: "510"; issue: "7504"; pages: "283-7"
COMMENT     Sequence Label: pMSCV-SMYD3
FEATURES             Location/Qualifiers
     source          1..8695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..40
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(78..100)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     200..219
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     413..435
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     563..582
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             782..1298
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1362..1703
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1770..2186
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2116..2138
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2154..2170
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2199..2208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2208..2231
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2259..2279
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2319..2348
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    2367..2391
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4084..4108)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4160..4183)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             4500..5513
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="ELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYVLR
                     VNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPETEL
                     PAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMD
                     DTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQ
                     YEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDD
                     AAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     LTR             5871..6385
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6547..6563
                     /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(6571..6577,6578..6595,6596..6601))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6616..6637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6754..6771)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6925..7513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7005..7024)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7684..8475,8476..8544))
                     /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     8307..8326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8545..8649)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
       61 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
      121 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
      181 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
      241 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
      301 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      361 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      421 cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
      481 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
      541 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
      601 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta
      661 aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc
      721 ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg
      781 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
      841 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
      901 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
      961 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     1021 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     1081 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     1141 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
     1201 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
     1261 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
     1321 agacccctgc ccagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
     1381 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
     1441 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     1501 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     1561 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     1621 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     1681 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     1741 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     1801 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     1861 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     1921 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     1981 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     2041 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     2101 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     2161 cagccctcac tccttctcta ggcgccggaa ttagatctgc cgccatggat tacaaggatg
     2221 acgacgataa ggattatgat attccaacta ctgctagcga gaatttgtat tttcagggtg
     2281 agctcaaaac cgcggctctt gcgcaacacg atgaagccga gcagaaactc atctctgaag
     2341 aggatctggg atccctcgag gttatcacaa gtttgtacaa aaaagcaggc tctttaaaga
     2401 tggagccgct gaaggtggaa aagttcgcaa ccgccaacag gggaaacggg ctgcgcgccg
     2461 tgaccccgct gcgccccgga gagctactct tccgctcgga tcccttggcg tacacggtgt
     2521 gcaaggggag tcgtggcgtc gtctgcgacc gctgccttct cgggaaggaa aagctgatgc
     2581 gatgctctca gtgccgcgtc gccaaatact gtagtgctaa gtgtcagaaa aaagcttggc
     2641 cagaccacaa gcgggaatgc aaatgcctta aaagctgcaa acccagatat cctccagact
     2701 ccgttcgact tcttggcaga gttgtcttca aacttatgga tggagcacct tcagaatcag
     2761 agaagcttta ctcattttat gatctggagt caaatattaa caaactgact gaagatagga
     2821 aagagggcct caggcaactc gtaatgacat ttcaacattt catgagagaa gaaatacagg
     2881 atgcctctca gctgccacct gcctttgacc tttttgaagc ctttgcaaaa gtgatctgca
     2941 actctttcac catctgtaat gcggagatgc aggaagttgg tgttggccta tatcccagta
     3001 tctctttgct caatcacagc tgtgacccca actgttcgat tgtgttcaat gggccccacc
     3061 tcttactgcg agcagtccga gacatcgagg tgggagagga gctcaccatc tgctacctgg
     3121 atatgctgat gaccagtgag gagcgccgga agcagctgag ggaccagtac tgctttgaat
     3181 gtgactgttt ccgttgccaa acccaggaca aggatgctga tatgctaact ggtgatgagc
     3241 aagtatggaa ggaagttcaa gaatccctga aaaaaattga agaactgaag gcacactgga
     3301 agtgggagca ggttctggcc atgtgccagg cgatcataag cagcaattct gaacggcttc
     3361 ccgatatcaa catctaccag ctgaaggtgc tcgactgcgc catggatgcc tgcatcaacc
     3421 tcggcctgtt ggaggaagcc ttgttctatg gtactcggac catggagcca tacaggattt
     3481 ttttcccagg aagccatccc gtcagagggg ttcaagtgat gaaagttggc aaactgcagc
     3541 tacatcaagg catgtttccc caagcaatga agaatctgag actggctttt gatattatga
     3601 gagtgacaca tggcagagaa cacagcctga ttgaagattt gattctactt ttagaagaat
     3661 gcgacgccaa catcagagca tcctaactcg agaaaaggat catggctata tagaaaattt
     3721 gacatttgga tgggatggac catcttggag gctactcaca gcccttaagt tgttatgtct
     3781 ggaagctgag aaatttacat gctggaaaaa agtacttctt ggggaggtaa tttcagatac
     3841 gaatgagaag acaagtttgg acatagccca gaaaatatgc tattatttca tagaagagac
     3901 taatgctgtg cttcaaaagg tgtctcatat gaaggatgaa aaagaggccc tgataaacca
     3961 actaactttg gtggaatcct tgtggacgga agagctaaag attctcaggg catctgccga
     4021 gaccctgcac agtttgcaaa cagcttttac ctgagaattc gcggccgcac tcgagatatc
     4081 tagacccagc tttcttgtac aaagtggtga taacgaattc taccgggtag gggaggcgct
     4141 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     4201 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     4261 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     4321 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     4381 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     4441 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggctg
     4501 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     4561 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     4621 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     4681 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
     4741 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     4801 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     4861 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     4921 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     4981 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     5041 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     5101 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     5161 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     5221 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     5281 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     5341 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     5401 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     5461 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagagtagat
     5521 gccgaccgaa caagagctga tttcgagaac gcctcagcca gcaactcgcg cgagcctagc
     5581 aaggcaaatg cgagagaacg gccttacgct tggtggcaca gttctcgtcc acagttcgct
     5641 aagctcgctc ggctgggtcg cgggagggcc ggtcgcagtg attcaggccc ttctggattg
     5701 tgttggtccc cagggcacga ttgtcatgcc cacgcactcg ggtgatctga ctgatcccgc
     5761 agattggaga tcgccgcccg tgcctgccga ttgggtgcag atccgtcgac ctgcagccaa
     5821 gcttatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     5881 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     5941 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     6001 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     6061 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     6121 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     6181 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     6241 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     6301 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     6361 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     6421 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     6481 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
     6541 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6601 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6661 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6721 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6781 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6841 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6901 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6961 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7021 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7081 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7141 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7201 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7261 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7321 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7381 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7441 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7501 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7561 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7621 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7681 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7741 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7801 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7861 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7921 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7981 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8041 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8101 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8161 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8221 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8281 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8341 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8401 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8461 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8521 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8581 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8641 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttatt
//
