LOCUS       sequence-170809-        7656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of shRNA targeting human SMYD3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7656)
  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 7656)
  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: pSico shSMYD3-C human
FEATURES             Location/Qualifiers
     source          1..7656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        428..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     581..601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             644..824
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    871..996
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1493..1726
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1911..1955
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2253..2370
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2623..2656)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2660..2680
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(2885..2908)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             3317..3916
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3317..3336)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3813..3833
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(4265..4298)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4354..4942
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4407..4427)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4825..4836)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4945..4961)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4946..4962)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5471..5651
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5713..6301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5793..5812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6472..7263,7264..7332))
                     /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     7095..7114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7333..7437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7512..7531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
       61 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      601 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
      661 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      721 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      781 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
      841 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
      901 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
      961 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     1021 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1081 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1141 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1201 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1261 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1321 accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     1381 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1441 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1501 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1561 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1621 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1681 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1741 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1801 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1861 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1921 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1981 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     2041 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     2101 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     2161 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     2221 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     2281 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     2341 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     2401 gtttggttag taccgggccc gctctagaga tccgacgcgc catctctagg cccgcgccgg
     2461 ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat
     2521 ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct
     2581 ttttaatact agctacattt tacatgatag gcttggattt ctataacttc gtatagcata
     2641 cattatacga agttataaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg
     2701 tgttttgaga ctataaatat cccttggaga aaagccttgt ttgcgtgtgt ctttgttgaa
     2761 tttcaagaga attcaacaaa gacacacgct tttttctcga gtactaggat ccattaggcg
     2821 gccgaattcg tcgacctcga aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     2881 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     2941 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     3001 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     3061 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     3121 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     3181 cgctttctgg gctcagaggc tgggaagggg tgggtccttc aaaagcgcac gtctgccgcg
     3241 ctgttctcct cttcctcatc tccgggcctt tcgacctgca tccatctaga tctcgatcga
     3301 gcagctgaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3361 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3421 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3481 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3541 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3601 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3661 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3721 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3781 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     3841 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     3901 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     3961 accccatgca tcgatgatat cataatttaa acaagcaaaa ccaaattaag ggccagctca
     4021 ttcctcccac tcatgatcta tagatctata gatctctcgt gggatcattg tttttctctt
     4081 gattcccact ttgtggttct aagtactgtg gtttccaaat gtgtcagttt catagcctga
     4141 agaacgagat cagcagcctc tgttccacat acacttcatt ctcagtattg ttttgccaag
     4201 ttctaattcc atcagaagct ggtcgactct agctagatgc ggccgcaatt cgtcgaggga
     4261 cctaataact tcgtatagca tacattatac gaagttatac atgtttaagg gttccggttc
     4321 cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     4381 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4441 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4501 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4561 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4621 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4681 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4741 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     4801 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4861 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4921 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
     4981 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     5041 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     5101 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5161 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     5221 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     5281 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     5341 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     5401 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     5461 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     5521 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5581 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     5641 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5881 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6121 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6181 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6961 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7021 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7081 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7141 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7201 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7261 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7321 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7381 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7441 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7501 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7561 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7621 ttgaccgaca attgcatgaa gaatctgctt agggtt
//
