LOCUS       sequence-170489-        8467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression of human SMYD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8467)
  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 8467)
  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
FEATURES             Location/Qualifiers
     source          1..8467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..27
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(65..87)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     187..206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     400..422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     550..569
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             769..1285
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1349..1690
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1757..2173
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2103..2125
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2141..2157
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2186..2195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2195..2218
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2246..2266
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2306..2335
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    2354..2378
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3727..3751)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3803..3826)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4162..4181
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4247..5272
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4247..5272
                     /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             5630..6144
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6306..6322
                     /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(6330..6336,6337..6354,6355..6360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6375..6396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6513..6530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6684..7272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6764..6783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7443..8234,8235..8303))
                     /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     8066..8085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8304..8408)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
       61 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
      121 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
      181 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
      241 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
      301 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      361 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      421 cgacggcgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
      481 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
      541 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
      601 gccggccacg atgcgtccgg cgtagaggcg attagtccaa tttgttaaag acaggatatc
      661 agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga
      721 gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
      781 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
      841 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
      901 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
      961 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     1021 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     1081 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
     1141 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata aagcctcttg
     1201 ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca gattgattga
     1261 ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga cccctgccca
     1321 gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc
     1381 tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt actagttagc
     1441 taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa cacccggccg
     1501 caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga cctgaggaag
     1561 ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag acgagaacct
     1621 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc
     1681 ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta
     1741 tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct taggtcactg
     1801 gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga gacgttgggt
     1861 taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag acggcacctt
     1921 taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc cgcatggaca
     1981 cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc cccctccctg
     2041 ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg ccccgtctct
     2101 cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag ccctcactcc
     2161 ttctctaggc gccggaatta gatctgccgc catggattac aaggatgacg acgataagga
     2221 ttatgatatt ccaactactg ctagcgagaa tttgtatttt cagggtgagc tcaaaaccgc
     2281 ggctcttgcg caacacgatg aagccgagca gaaactcatc tctgaagagg atctgggatc
     2341 cctcgaggtt atcacaagtt tgtacaaaaa agcaggctct ttaaaggaac caattcagtc
     2401 gactggatcc atgagggccg agggcctcgg cggcctggag cgcttctgca gcccgggcaa
     2461 aggccggggg ctgcgggctc tgcagccctt ccaggtgggg gacttgctgt tctcctgccc
     2521 ggcctatgcc tacgtgctca cggtcaacga gcggggcaac cactgcgagt actgcttcac
     2581 caggaaagaa ggattgtcca aatgtggaag atgcaagcag gcattttact gcaatgtgga
     2641 gtgtcagaaa gaagattggc ccatgcacaa gctggaatgt tctcccatgg ttgtttttgg
     2701 ggaaaactgg aatccctcgg agactgtaag actaacagca aggattctgg ccaaacagaa
     2761 aatccaccca gagagaacac cttcggaaaa attgttagct gtgaaggagt ttgaatcaca
     2821 tctggataag ttagacaatg agaagaagga tttgattcag agtgacatag ctgctctcca
     2881 tcacttttac tccaagcatc tcgaattccc tgacaatgat agcctcgtag tactctttgc
     2941 acaggttaac tgtaatggct tcacaattga agatgaagaa ctttctcatt tgggatcagc
     3001 gatatttcct gatgttgcat tgatgaatca tagctgttgc cccaatgtca ttgtgaccta
     3061 caaagggacc ctggcagaag tcagagctgt acaggaaatc aagccgggag aggaggtttt
     3121 taccagctat attgatctcc tgtacccaac ggaagataga aatgaccggt taagagattc
     3181 ttatttcttt acctgtgagt gccaggagtg taccaccaag gacaaggata aggccaaggt
     3241 ggaaatccgg aagctcagcg atcccccaaa ggcagaagcc atccgagaca tggtcagata
     3301 tgcacgcaac gtcattgaag agttccggag ggccaagcac tataaatccc ctagtgagct
     3361 gctggagatc tgcgagctca gccaggagaa gatgagctct gtgtttgagg acagtaacgt
     3421 gtacatgttg cacatgatgt accaggccat gggtgtctgc ttgtacatgc aggactggga
     3481 aggagccctg caatatggac agaaaatcat taagccctac agtaagcact atcctttgta
     3541 ctccctcaac gtggcctcca tgtggttgaa gctagggaga ctctacatgg gcctggaaca
     3601 caaagccgca ggggagaaag ccctgaagaa ggccattgca atcatggaag tagctcacgg
     3661 caaagatcat ccatatattt ctgagatcaa acaggaaatt gaaagccact gactcgagat
     3721 atctagaccc agctttcttg tacaaagtgg tgataacgaa ttctaccggg taggggaggc
     3781 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     3841 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     3901 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     3961 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     4021 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     4081 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggccc
     4141 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     4201 tctcctcttc ctcatctccg ggcctttcga cctgcatccc gccaccatga aaaagcctga
     4261 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     4321 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     4381 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     4441 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     4501 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     4561 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     4621 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     4681 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     4741 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4801 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4861 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4921 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4981 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     5041 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     5101 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     5161 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     5221 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agagtagatg
     5281 ccgaccgaac aagagctgat ttcgagaacg cctcagccag caactcgcgc gagcctagca
     5341 aggcaaatgc gagagaacgg ccttacgctt ggtggcacag ttctcgtcca cagttcgcta
     5401 agctcgctcg gctgggtcgc gggagggccg gtcgcagtga ttcaggccct tctggattgt
     5461 gttggtcccc agggcacgat tgtcatgccc acgcactcgg gtgatctgac tgatcccgca
     5521 gattggagat cgccgcccgt gcctgccgat tgggtgcaga tccgtcgacc tgcagccaag
     5581 cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     5641 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
     5701 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     5761 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     5821 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     5881 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     5941 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
     6001 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg
     6061 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     6121 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct
     6181 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca
     6241 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg
     6301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8401 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8461 acattaa
//
