LOCUS       sequence-372801-        7615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of shRNA targeting Cavin1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7615)
  AUTHORS   Teo JL, Gomez GA, Weeratunga S, Davies EM, Noordstra I, Budnar S, 
            Katsuno-Kambe H, McGrath MJ, Verma S, Tomatis V, Acharya BR, 
            Balasubramaniam L, Templin RM, McMahon KA, Lee YS, Ju RJ, Stebhens 
            SJ, Ladoux B, Mitchell CA, Collins BM, Parton RG, Yap AS
  TITLE     Caveolae Control Contractile Tension for Epithelia to Eliminate 
            Tumor Cells.
  JOURNAL   Dev Cell. 2020 Jul 6;54(1):75-91.e7. doi: 
            10.1016/j.devcel.2020.05.002. Epub 2020 Jun 1.
  PUBMED    32485139
REFERENCE   2  (bases 1 to 7615)
  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: 
            "10.1016/j.devcel.2020.05.002"; journalName: "Dev Cell"; date: 
            "2020-07-6- 6"; volume: "54"; issue: "1"; pages: "75-91.e7"
COMMENT     Sequence Label: pLL5.0 ShCavin1-EGFP
FEATURES             Location/Qualifiers
     source          1..7615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(72..91)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        263..642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        644..842
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     797..817
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             860..1040
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1087..1212
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1709..1942
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2127..2171
                     /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    2469..2586
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2641..2953
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2872..2892
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     3010..3026
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3068..3101)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3689..3698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3695..3697,3698..3700,3701..4414)
                     /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(3695..3697,3698..3700,3701..4411)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3740..3761)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4001..4020)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4348..4369
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4442..4475)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4531..5117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4584..4604)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5002..5013)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5120..5136)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5646..5826
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5888..6476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5968..5987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6647..7438,7439..7507))
                     /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     7270..7289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7508..7612)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
       61 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      121 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      181 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      241 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      301 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      361 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      421 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      481 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      541 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      601 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      661 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      721 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      781 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      841 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
      901 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
      961 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     1021 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     1081 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     1141 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     1201 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     1261 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     1321 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     1381 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     1441 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     1501 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccggccgc
     1561 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1621 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1681 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1741 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1801 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1861 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1921 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1981 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     2041 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     2101 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     2161 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     2221 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     2281 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     2341 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     2401 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     2461 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2521 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2581 aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc gcgctctaga
     2641 gatccgacgc gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc
     2701 tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct
     2761 taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat
     2821 aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa
     2881 aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga
     2941 gaaaagcctt gttttgagca gccggaccaa taattcaaga gattattggt ccggctgctc
     3001 attttttctc gaggtcgacg gtatcgataa gctcgcttca cgagattcca gcaggtcgag
     3061 ggacctaata acttcgtata gcatacatta tacgaagtta tattaagggt tccaagctta
     3121 agcggccgct gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     3181 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     3241 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     3301 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     3361 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     3421 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     3481 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgta
     3541 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg
     3601 gtctcctcag attgattgac tgcccacctc gggggtcttt catcgaattc tgcagtcgac
     3661 ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     3721 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3781 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3841 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3901 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3961 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4021 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4081 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4141 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4201 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4261 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4321 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4381 gatcactctc ggcatggacg agctgtacaa gtaacctgca ggcaattcgt cgagggacct
     4441 aataacttcg tatagcatac attatacgaa gttatacatg tttaagggtt ccggttccac
     4501 taggtacaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     4561 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4621 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4681 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4741 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4801 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4861 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4921 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4981 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccggcctgct
     5041 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5101 ttgggccgcc tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag
     5161 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     5221 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     5281 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     5341 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     5401 ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt
     5461 gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca
     5521 cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag
     5581 agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact
     5641 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5701 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5761 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5821 ctagcagcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5881 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5941 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6001 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6061 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6121 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6181 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6241 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6301 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6361 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6421 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6481 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6541 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6601 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6661 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6721 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6781 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6841 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6901 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6961 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7021 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7081 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7141 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7201 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7261 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7321 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7381 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7441 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7501 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7561 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcac
//
