LOCUS       sequence-459543-        8813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of shRNA targeting Cavin1 + reconstitution of 
            mouse Cavin1 (Cavin1 rescue).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8813)
  AUTHORS   Brooks JW, Tillu V, Eckert J, Verma S, Collins BM, Parton RG, Yap AS
  TITLE     Caveola mechanotransduction reinforces the cortical cytoskeleton to 
            promote epithelial resilience.
  JOURNAL   Mol Biol Cell. 2023 Nov 1;34(12):ar120. doi: 
            10.1091/mbc.E23-05-0163. Epub 2023 Sep 6.
  PUBMED    37672337
REFERENCE   2  (bases 1 to 8813)
  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: "Mol Biol 
            Cell."; date: "2023-11-1"; volume: "34(12):ar120. doi"; pages: " 
            10.1091/mbc.E23-05-0163. Epub 2023 Sep 6"
COMMENT     Sequence Label: pLL5.0_shRNAHsCavin1_MmCavin1_EGFP
FEATURES             Location/Qualifiers
     source          1..8813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2616..2929
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2848..2868
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     3006..3022
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3064..3097)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3668..3677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4862..4871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4868..4870,4871..4873,4874..5587)
                     /codon_start=1
                     /product="blue variant of GFP"
                     /label=BFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4913..4934)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5174..5193)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5521..5542
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5615..5648)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5704..6290
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5757..5777)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6175..6186)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6293..6309)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6819..6999
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7061..7649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7141..7160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7820..8611,8612..8680))
                     /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     8443..8462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8681..8785)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgcgct ctagagatcc gacgccgcca tctctaggcc
     2641 cgcgccggcc ccctcgcaca gacttgtggg agaagctcgg ctactcccct gccccggtta
     2701 atttgcatat aatatttcct agtaactata gaggcttaat gtgcgataaa agacagataa
     2761 tctgttcttt ttaatactag ctacatttta catgataggc ttggatttct ataagagata
     2821 caaatactaa attattattt taaaaaacag cacaaaagga aactcaccct aactgtaaag
     2881 taattgtgtg ttttgagact ataaatatcc cttggagaaa agccttgttt caccttccac
     2941 gtcaagaaga tccgcgaggt tcaagagacc tcgcggatct tcttgacgtg gaaggtgttt
     3001 tttctcgagg tcgacggtat cgataagctc gcttcacgag attccagcag gtcgagggac
     3061 ctaataactt cgtatagcat acattatacg aagttatatt aagggttcca agcttaagcg
     3121 gccgctgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     3181 ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg aacagagaga
     3241 cagcagaata tgggccaaca aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     3301 caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga
     3361 tgtttccagg gtgccccaag gacctgaaaa tgaccctgtg ccttatttga actaaccaat
     3421 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     3481 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
     3541 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
     3601 tcctcagatt gattgactgc ccacctcggg ggtctttcat cgaattctgc agtcgacggt
     3661 accgcgggcc accatggagg atgtcacgct ccatatcgtt gagcggccgt attccggatt
     3721 tcccgatgct tcctcagagg gcccggagcc cacccaaggg gaggcgcggg ccacggagga
     3781 gccgtcgggg accggctccg acgagctgat caagtcggac caggtgaacg gtgtgctggt
     3841 gctgagcctt ctggataaaa tcatcggcgc cgttgaccag atccagctga cccaagccca
     3901 gctggaggag cgacaggcgg agatggaggg cgctgtgcag agcatccagg gagagctgag
     3961 caagctgggc aaggcgcacg ccaccacgag caacaccgtg agcaagttgc tggagaaggt
     4021 gcgcaaggtc agcgtcaacg tgaagaccgt gcgcggcagc ctggagcgcc aggccggcca
     4081 gataaagaaa ctggaggtca acgaggcgga gctgctgagg cgccgcaact tcaaagtcat
     4141 gatctaccag gatgaagtca agctgccggc caaactgagc gtcagcaagt cgctgaaaga
     4201 gtcggaggca ctgcctgaga aggagggtga cgagctgggc gagggcgagc ggcccgagga
     4261 tgacaccgcg gcgatcgagc tgtcgtccga cgaggcggtg gaggtggagg aggtgatcga
     4321 ggagtcccgc gccgagcgca tcaagcgcag cggcctgcgg cgcgtggacg acttcaagaa
     4381 ggccttctcc aaggagaaga tggagaagac caaggtgcgc acgcgtgaga acctggagaa
     4441 gacgcgcctg aagaccaagg agaacctgga gaagacacgg cacacgctgg agaagcgcat
     4501 gaacaagctg ggcacgcgcc tggtgcccgt ggagcgacga gagaagctga agacatcccg
     4561 ggacaagctg cgcaagtcct tcacgcccga ccatgtggtg tatgcgcgct ccaagaccgc
     4621 tgtctacaag gtgccgcctt tcacatttca tgttaagaaa attcgtgaag gcgaggtgga
     4681 ggtgctgaag gccaccgaga tggtggaggt gggtcccgag gacgacgagg ttggcgcgga
     4741 gcgcggcgag gccactgacc tgctgcgcgg gagcagcccc gacgtgcaca cgctgctgga
     4801 gatcaccgag gagtcggacg ccgtcctggt ggacaagagc gacagcgacg atccaccggt
     4861 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4921 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     4981 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     5041 gcccaccctc gtgaccaccc tgagccacgg cgtccagtgc ttcagccgct accccgacca
     5101 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     5161 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     5221 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     5281 ggggcacaag ctggagtaca acttcaacag ccacaacgtc tatatcatgg ccgacaagca
     5341 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     5401 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     5461 cagccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     5521 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     5581 caagtaacct gcaggcaatt cgtcgaggga cctaataact tcgtatagca tacattatac
     5641 gaagttatac atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc
     5701 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5761 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5821 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5881 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5941 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     6001 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     6061 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     6121 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     6181 ctcaatccag cggaccttcc ttcccggcct gctgccggct ctgcggcctc ttccgcgtct
     6241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     6301 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     6361 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     6421 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6481 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     6541 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     6601 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     6661 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     6721 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     6781 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
     6841 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6901 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6961 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc
     7021 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7081 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7141 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7201 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7261 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7321 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7381 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7441 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7501 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7561 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7681 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7741 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7801 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7861 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7921 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7981 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8041 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8101 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8161 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8221 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8281 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8341 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8401 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8461 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8521 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8581 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8641 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8701 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8761 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
