LOCUS       sequence-170816-        7627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of shRNA targeting human MAPKAPK3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7627)
  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 7627)
  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: pSico shMAPKAPK3 human
FEATURES             Location/Qualifiers
     source          1..7627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        491..689
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     644..664
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             707..887
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    934..1059
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1556..1789
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1974..2018
                     /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    2316..2433
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2686..2719)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2723..2743
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(2948..2971)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             3351..3950
                     /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(3351..3370)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3847..3867
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(4299..4332)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4388..4976
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4441..4461)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4859..4870)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4979..4995)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4980..4996)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5505..5685
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5747..6335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5827..5846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6506..7297,7298..7366))
                     /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     7129..7148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7367..7471)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7546..7565)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
       61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      661 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
      721 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
      781 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
      841 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
      901 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
      961 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     1021 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     1081 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     1141 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     1201 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     1261 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     1321 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     1381 accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg
     1441 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1501 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1561 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1621 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1681 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1741 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1801 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1861 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1921 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1981 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     2041 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     2101 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     2161 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     2221 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
     2281 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     2341 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     2401 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     2461 ccagtttggt tagtaccggg cccgctctag agatccgacg cgccatctct aggcccgcgc
     2521 cggccccctc gcacagactt gtgggagaag ctcggctact cccctgcccc ggttaatttg
     2581 catataatat ttcctagtaa ctatagaggc ttaatgtgcg ataaaagaca gataatctgt
     2641 tctttttaat actagctaca ttttacatga taggcttgga tttctataac ttcgtatagc
     2701 atacattata cgaagttata aacagcacaa aaggaaactc accctaactg taaagtaatt
     2761 gtgtgttttg agactataaa tatcccttgg agaaaagcct tgtttgctaa gtggcttccc
     2821 attattcaag agataatggg aagccactta gcttttttct cgagtactag gatccattag
     2881 gcggccgaat tcgtcgacct cgaaattcta ccgggtaggg gaggcgcttt tcccaaggca
     2941 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     3001 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     3061 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     3121 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     3181 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     3241 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggttcct catctccggg
     3301 cctttcgacc tgcatccatc tagatctcga tcgagcagct gaagcttacc atgaccgagt
     3361 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     3421 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     3481 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     3541 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     3601 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     3661 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     3721 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     3781 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     3841 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     3901 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc
     3961 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcgatg atatcataat
     4021 ttaaacaagc aaaaccaaat taagggccag ctcattcctc ccactcatga tctatagatc
     4081 tatagatctc tcgtgggatc attgtttttc tcttgattcc cactttgtgg ttctaagtac
     4141 tgtggtttcc aaatgtgtca gtttcatagc ctgaagaacg agatcagcag cctctgttcc
     4201 acatacactt cattctcagt attgttttgc caagttctaa ttccatcaga agctggtcga
     4261 ctctagctag atgcggccgc aattcgtcga gggacctaat aacttcgtat agcatacatt
     4321 atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct
     4381 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4441 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4501 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4561 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4621 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4681 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4741 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4801 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4861 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4921 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4981 ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     5041 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     5101 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5161 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     5221 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     5281 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     5341 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     5401 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     5461 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     5521 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5581 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5641 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
     5701 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5761 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5821 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5881 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5941 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6001 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6061 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7501 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     7561 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     7621 cgcgagc
//
