LOCUS       sequence-310643-        8386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown of Rictor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8386)
  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 8386)
  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-Rictor-shRNA
FEATURES             Location/Qualifiers
     source          1..8386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     232..252
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     374..390
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(432..465)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        581..884
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        885..1088
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             1117..2547
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(2567..2600)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2656..3244
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2709..2729)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3127..3138)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3247..3263)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             3773..3953
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4015..4603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4095..4114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4774..5565,5566..5634))
                     /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     5397..5416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5635..5739)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5814..5833)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6005..6384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6386..6584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6602..6782
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6829..6954
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7451..7684
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7869..7913
                     /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    8211..8328
                     /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 gtttgccagt aagatgggaa tcattttcaa gagaaatgat tcccatctta
      361 ctggcttttt tctcgaggtc gacggtatcg ataagctcgc ttcacgagat tccagcaggt
      421 cgagggacct aataacttcg tatagcatac attatacgaa gttatattaa gggttccaag
      481 cttaagcggc cgcgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
      541 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      601 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      661 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      721 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      781 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      841 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      901 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      961 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     1021 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1081 gcagagctgg tttagtgaac cgtcagatcc gctagcatgg tgagcaaggg cgaggaggtc
     1141 atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc
     1201 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1261 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac
     1321 ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc
     1381 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg
     1441 acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc
     1501 aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc
     1561 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg
     1621 aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg
     1681 caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac
     1741 tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat
     1801 ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg
     1861 gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac
     1921 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     1981 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     2041 atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg
     2101 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg
     2161 accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc
     2221 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     2281 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg
     2341 aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag
     2401 cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac
     2461 gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg
     2521 tacggcatgg acgagctgta caagtaagaa ttcgtcgagg gacctaataa cttcgtatag
     2581 catacattat acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat
     2641 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2701 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2761 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2821 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2881 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2941 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3001 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3061 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3121 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3181 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3241 ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag
     3301 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
     3361 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
     3421 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     3481 acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa
     3541 ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt
     3601 accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca
     3661 ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga
     3721 cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct
     3781 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     3841 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     3901 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg
     3961 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4021 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4081 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4141 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4201 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4261 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4321 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4381 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4441 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4501 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4561 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4621 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4681 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4741 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4801 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4861 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4921 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4981 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5041 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5101 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5161 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5221 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5281 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5341 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5401 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5461 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5521 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5581 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5641 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5701 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5761 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     5821 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     5881 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     5941 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     6001 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     6061 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     6121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     6181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     6241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     6301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     6361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     6421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     6481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     6541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac
     6601 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6661 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6721 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6781 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     6841 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     6901 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     6961 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     7021 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     7081 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     7141 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     7201 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     7261 caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag
     7321 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     7381 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     7441 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     7501 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     7561 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     7621 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     7681 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     7741 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     7801 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     7861 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     7921 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     7981 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     8041 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     8101 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     8161 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     8221 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     8281 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     8341 agggacagca gagatccagt ttggttagta ccgggcccgc tctaga
//
