LOCUS       sequence-170488-        8394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of human SMYD2 F184A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8394)
  AUTHORS   Reynoird N, Mazur PK, Stellfeld T, Flores NM, Lofgren SM, Carlson 
            SM, Brambilla E, Hainaut P, Kaznowska EB, Arrowsmith CH, Khatri P, 
            Stresemann C, Gozani O, Sage J
  TITLE     Coordination of stress signals by the lysine methyltransferase SMYD2
            promotes pancreatic cancer.
  JOURNAL   Genes Dev. 2016 Apr 1;30(7):772-85. doi: 10.1101/gad.275529.115. 
            Epub 2016 Mar 17.
  PUBMED    26988419
REFERENCE   2  (bases 1 to 8394)
  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: 
            "10.1101/gad.275529.115"; journalName: "Genes Dev"; date: 
            "2016-04-1- 1"; volume: "30"; issue: "7"; pages: "772-85"
COMMENT     Sequence Label: pMSCV-SMYD2 F184A
FEATURES             Location/Qualifiers
     source          1..8394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..67)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     167..186
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     380..402
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     530..549
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             749..1265
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1329..1670
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1737..2153
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2083..2105
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2121..2137
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2166..2175
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2175..2198
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2226..2246
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2286..2315
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    2334..2358
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3707..3731)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3783..3806)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             4154..5179
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4154..5179
                     /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             5537..6051
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6213..6229
                     /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(6237..6243,6244..6261,6262..6267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6282..6303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6420..6437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6591..7179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6671..6690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7350..8141,8142..8210))
                     /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     7973..7992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8211..8315)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(8383..8394,1..7)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
       61 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
      121 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
      181 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
      241 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
      301 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
      361 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg
      421 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
      481 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
      541 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
      601 cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt
      661 tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga
      721 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
      781 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
      841 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
      901 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
      961 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     1021 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     1081 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     1141 tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
     1201 gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct cgggggtctt
     1261 tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg acccccccgc
     1321 cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg
     1381 ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     1441 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     1501 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     1561 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     1621 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     1681 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     1741 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1801 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1861 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1921 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1981 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     2041 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     2101 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaatta
     2161 gatctgccgc catggattac aaggatgacg acgataagga ttatgatatt ccaactactg
     2221 ctagcgagaa tttgtatttt cagggtgagc tcaaaaccgc ggctcttgcg caacacgatg
     2281 aagccgagca gaaactcatc tctgaagagg atctgggatc cctcgaggtt atcacaagtt
     2341 tgtacaaaaa agcaggctct ttaaaggaac caattcagtc gactggatcc atgagggccg
     2401 agggcctcgg cggcctggag cgcttctgca gcccgggcaa aggccggggg ctgcgggctc
     2461 tgcagccctt ccaggtgggg gacttgctgt tctcctgccc ggcctatgcc tacgtgctca
     2521 cggtcaacga gcggggcaac cactgcgagt actgcttcac caggaaagaa ggattgtcca
     2581 aatgtggaag atgcaagcag gcattttact gcaatgtgga gtgtcagaaa gaagattggc
     2641 ccatgcacaa gctggaatgt tctcccatgg ttgtttttgg ggaaaactgg aatccctcgg
     2701 agactgtaag actaacagca aggattctgg ccaaacagaa aatccaccca gagagaacac
     2761 cttcggaaaa attgttagct gtgaaggagt ttgaatcaca tctggataag ttagacaatg
     2821 agaagaagga tttgattcag agtgacatag ctgctctcca tcacttttac tccaagcatc
     2881 tcgaattccc tgacaatgat agcctcgtag tactctttgc acaggttaac tgtaatggcg
     2941 ccacaattga agatgaagaa ctttctcatt tgggatcagc gatatttcct gatgttgcat
     3001 tgatgaatca tagctgttgc cccaatgtca ttgtgaccta caaagggacc ctggcagaag
     3061 tcagagctgt acaggaaatc aagccgggag aggaggtttt taccagctat attgatctcc
     3121 tgtacccaac ggaagataga aatgaccggt taagagattc ttatttcttt acctgtgagt
     3181 gccaggagtg taccaccaag gacaaggata aggccaaggt ggaaatccgg aagctcagcg
     3241 atcccccaaa ggcagaagcc atccgagaca tggtcagata tgcacgcaac gtcattgaag
     3301 agttccggag ggccaagcac tataaatccc ctagtgagct gctggagatc tgcgagctca
     3361 gccaggagaa gatgagctct gtgtttgagg acagtaacgt gtacatgttg cacatgatgt
     3421 accaggccat gggtgtctgc ttgtacatgc aggactggga aggagccctg caatatggac
     3481 agaaaatcat taagccctac agtaagcact atcctttgta ctccctcaac gtggcctcca
     3541 tgtggttgaa gctagggaga ctctacatgg gcctggaaca caaagccgca ggggagaaag
     3601 ccctgaagaa ggccattgca atcatggaag tagctcacgg caaagatcat ccatatattt
     3661 ctgagatcaa acaggaaatt gaaagccact gactcgagat atctagaccc agctttcttg
     3721 tacaaagtgg tgataacgaa ttctaccggg taggggaggc gcttttccca aggcagtctg
     3781 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     3841 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     3901 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     3961 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     4021 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     4081 ctttctgggc tcagaggctg ggaaggggtg ggtcttcctc atctccgggc ctttcgacct
     4141 gcatcccgcc accatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     4201 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4261 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4321 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     4381 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4441 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4501 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4561 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4621 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4681 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4741 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4801 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4861 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4921 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4981 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     5041 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     5101 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     5161 tccgagggca aaggaataga gtagatgccg accgaacaag agctgatttc gagaacgcct
     5221 cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt
     5281 ggcacagttc tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc
     5341 gcagtgattc aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg
     5401 cactcgggtg atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg
     5461 gtgcagatcc gtcgacctgc agccaagctt atcgataaaa taaaagattt tatttagtct
     5521 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     5581 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     5641 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5701 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag
     5761 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     5821 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     5881 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg
     5941 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     6001 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag
     6061 tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac
     6121 tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga
     6181 cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac
     6241 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6301 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7021 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7261 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7321 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7381 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7441 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7501 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7561 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7621 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7681 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7741 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7801 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7861 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7921 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7981 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8041 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8101 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8161 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8221 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8281 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8341 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatc
//
