LOCUS       sequence-310630-        7687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown of caspase-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7687)
  AUTHORS   Xu ZX, Tan JW, Xu H, Hill CJ, Ostrovskaya O, Martemyanov KA, Xu B
  TITLE     Caspase-2 promotes AMPA receptor internalization and cognitive 
            flexibility via mTORC2-AKT-GSK3beta signaling.
  JOURNAL   Nat Commun. 2019 Aug 9;10(1):3622. doi: 10.1038/s41467-019-11575-1.
  PUBMED    31399584
REFERENCE   2  (bases 1 to 7687)
  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"; journalName: "Nat 
            Commun."; date: "2019-08-9"; volume: "10(1):3622. doi"; pages: " 
            10.1038/s41467-019-11575-1"
COMMENT     Sequence Label: pLL3.7-Casp2-shRNA
FEATURES             Location/Qualifiers
     source          1..7687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     232..252
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     370..386
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(428..461)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        577..880
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        881..1084
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1123..1132
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1129..1131,1132..1134,1135..1848)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1129..1131,1132..1134,1135..1845)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1174..1195)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1435..1454)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1782..1803
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1868..1901)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1957..2545
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2010..2030)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2428..2439)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2548..2564)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             3074..3254
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3316..3904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3396..3415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4075..4866,4867..4935))
                     /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     4698..4717
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4936..5040)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5115..5134)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5306..5685
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5687..5885
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5903..6083
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6130..6255
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6752..6985
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7170..7214
                     /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    7512..7629
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gatccgacgc gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc
       61 tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct
      121 taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat
      181 aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa
      241 aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga
      301 gaaaagcctt gtttgccaga atgtggaact cctttcaaga gaaggagttc cacattctgg
      361 cttttttctc gaggtcgacg gtatcgataa gctcgcttca cgagattcca gcaggtcgag
      421 ggacctaata acttcgtata gcatacatta tacgaagtta tattaagggt tccaagctta
      481 agcggccgcg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
      541 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      601 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      661 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      721 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      781 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      841 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      901 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      961 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     1021 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     1081 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag
     1141 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1201 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1261 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1321 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1381 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1441 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1501 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1561 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1621 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1681 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1741 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1801 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagga attcgtcgag
     1861 ggacctaata acttcgtata gcatacatta tacgaagtta tacatgttta agggttccgg
     1921 ttccactagg tacaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     1981 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2041 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2101 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2161 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2221 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2281 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2341 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     2401 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2461 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2521 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgatcgag acctagaaaa
     2581 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     2641 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     2701 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     2761 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     2821 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     2881 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     2941 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     3001 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     3061 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     3121 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     3181 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     3241 gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3301 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3361 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3421 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3481 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3541 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3601 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3661 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3721 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3781 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3841 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3901 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3961 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4021 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4081 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4141 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4201 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4261 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4321 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4381 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4441 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4501 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4561 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4621 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4681 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4741 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4801 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4861 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4921 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4981 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5041 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     5101 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     5161 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     5221 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     5281 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     5341 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     5401 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     5461 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     5521 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     5581 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     5641 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     5701 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     5761 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     5821 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     5881 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     5941 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6001 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6061 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     6121 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     6181 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     6241 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     6301 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     6361 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     6421 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     6481 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     6541 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc
     6601 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     6661 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     6721 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     6781 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     6841 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     6901 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     6961 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     7021 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     7081 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     7141 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     7201 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     7261 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     7321 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     7381 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     7441 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
     7501 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     7561 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     7621 caaaattttc gggtttatta cagggacagc agagatccag tttggttagt accgggcccg
     7681 ctctaga
//
