LOCUS       sequence-459542-        8999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of shRNA targeting Cavin1 + reconstitution of 
            zebrafish Cavin1 (Cavin1 rescue).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8999)
  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 8999)
  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_DrCavin1b_EGFP
FEATURES             Location/Qualifiers
     source          1..8999
                     /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      5048..5057
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5054..5056,5057..5059,5060..5773)
                     /codon_start=1
                     /product="blue variant of GFP"
                     /label=BFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5099..5120)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5360..5379)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5707..5728
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5801..5834)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5890..6476
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5943..5963)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6361..6372)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6479..6495)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7005..7185
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7247..7835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7327..7346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8006..8797,8798..8866))
                     /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     8629..8648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8867..8971)
                     /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 tgattgaaga cagccgtctg catctggagg tgcccgagct
     3721 gcgcaagtct cttactgaga tctcagaaga tgaaatcaac ctcccggtct ctgatgatga
     3781 agacgaagcc accgccgaga acaaagctaa agatgctgcc gagaaagaag aggctgggga
     3841 gggtggcgag atgaatgggg tcatggtcct cacgctgctt gacaaaatca ttggagccgt
     3901 tgaccagatc cagcagacgc aggctgggtt ggaagcccgt cagcgggaca tggagagatc
     3961 tgtgacggga atccaggggg agctcaccaa actgtccaag agccacacca ccacagccaa
     4021 caccgtcaat aagatgctgg agaaggtgcg taaggtgagc gtgaacgtca agtcggtgcg
     4081 ggggaacctg gagaaacagg ctggccagat caaacgtctg gagagcaacg agaacgagct
     4141 cctaaagaga agaaacttta aagtcatgat ctatcaggat gaagtaaaag tgggttcgaa
     4201 agtgaatgtg gccaagtcta tgaagtatcc agagcctgtg gaaggggaag ctgagggtga
     4261 actcaaagct gcagcagaag aaggaggaga ggacaaagag aaagttaatc tggacctgtc
     4321 ctctgatgaa gaggaggtgg agattgagga gacaattgag gagtctcgag cagaacgcat
     4381 caagcggagc agcctgcaac gtgtccagaa catcaaaacc gttttctcca aggggaatat
     4441 ggagaaaacc aaacagaaga ccaaggagaa cttggagaag accaaacaga agaccaaaga
     4501 gaacctggag aaaactcgca tgaggaccaa agagaacttg gaaaaaacaa gacagaagac
     4561 taaggaaaac cttgaaaaga ccaaacagaa gacccgagag aacctggaga aaacacgtca
     4621 caacatagaa aagaaaatgg gtaaacttgg cacccgcatg tccgtgaacc ccgagcgcaa
     4681 agagaagatg aagacttcac agaagaaggt gatgaagtct tttactcctg accacaccat
     4741 ctacgcccgc tccaaaacgg ccgtctacaa ggttccaccg ttcacatttc atgtgaagaa
     4801 aattcgtgaa ggagaagtgg agattcaggg aactgagatg gtggaagttg tcggcgacac
     4861 agaggaggtg gagaacgcag tgatggaggg agaggaagat gcagagatgg agatggaagc
     4921 tggagagatg gtggaggaag acgaggacga gacagaagac agccctgaga tgagagcgct
     4981 gctggagaaa ggagacgagc tggtgctggt ggaagatcat gacagagaca gcgatgatcc
     5041 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     5101 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     5161 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     5221 gccctggccc accctcgtga ccaccctgag ccacggcgtc cagtgcttca gccgctaccc
     5281 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     5341 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     5401 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     5461 catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata tcatggccga
     5521 caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag
     5581 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     5641 gcccgacagc cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     5701 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     5761 gctgtacaag taacctgcag gcaattcgtc gagggaccta ataacttcgt atagcataca
     5821 ttatacgaag ttatacatgt ttaagggttc cggttccact aggtacaatt cgatatcaag
     5881 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5941 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     6001 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     6061 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     6121 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     6181 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     6241 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     6301 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     6361 tcggccctca atccagcgga ccttccttcc cggcctgctg ccggctctgc ggcctcttcc
     6421 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     6481 ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     6541 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     6601 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6661 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     6721 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     6781 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     6841 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     6901 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     6961 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     7021 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7081 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7141 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
     7201 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7261 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7321 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7381 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7441 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7501 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7561 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7621 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7681 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7741 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7921 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7981 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8101 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8161 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8281 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8881 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8941 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
