LOCUS       sequence-373164-        8018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of shRNA targeting Caveolin1 and NLS-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8018)
  AUTHORS   Katsuno-Kambe H, Parton RG, Yap AS, Teo JL
  TITLE     Caveolin-1 influences epithelial collective cell migration via FMNL2
            formin.
  JOURNAL   Biol Cell. 2021 Feb;113(2):107-117. doi: 10.1111/boc.202000116. Epub
            2020 Nov 25.
  PUBMED    33169848
REFERENCE   2  (bases 1 to 8018)
  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: 
            "10.1111/boc.202000116"; journalName: "Biol Cell"; date: "2021-02"; 
            volume: "113"; issue: "2"; pages: "107-117"
COMMENT     Sequence Label: pLL5.0 shCAV1-NLS-mCherry
FEATURES             Location/Qualifiers
     source          1..8018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             844..1024
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1071..1196
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1693..1926
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2111..2155
                     /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    2453..2570
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2625..2937
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2856..2876
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2994..3010
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3052..3085)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3655..4032
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     regulatory      4048..4057
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4199..4217)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4467..4486
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(4829..4862)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4918..5504
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4971..4991)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5389..5400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5507..5523)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6033..6213
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6275..6863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6355..6374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7034..7825,7826..7894))
                     /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     7657..7676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7895..7999)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
     1561 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     1621 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     1681 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     1741 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     1801 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     1861 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     1921 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     1981 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     2041 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     2101 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     2161 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     2221 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     2281 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     2341 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     2401 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     2461 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2521 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     2581 acagggacag cagagatcca gtttggttag taccgcgctc tagagatccg acgcgccatc
     2641 tctaggcccg cgccggcccc ctcgcacaga cttgtgggag aagctcggct actcccctgc
     2701 cccggttaat ttgcatataa tatttcctag taactataga ggcttaatgt gcgataaaag
     2761 acagataatc tgttcttttt aatactagct acattttaca tgataggctt ggatttctat
     2821 aagagataca aatactaaat tattatttta aaaaacagca caaaaggaaa ctcaccctaa
     2881 ctgtaaagta attgtgtgtt ttgagactat aaatatccct tggagaaaag ccttgtttgc
     2941 atctttctct ctgttaattc aagagattaa cagagagaaa gatgcttttt tctcgaggtc
     3001 gacggtatcg ataagctcgc ttcacgagat tccagcaggt cgagggacct aataacttcg
     3061 tatagcatac attatacgaa gttatattaa gggttccaag cttaagcggc cgctgaaaga
     3121 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
     3181 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
     3241 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     3301 gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     3361 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     3421 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     3481 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
     3541 ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc tcagattgat
     3601 tgactgccca cctcgggggt ctttcatcga attctgcagt cgacggtacc gcggatgcca
     3661 gagccagcga agtctgctcc cgccccgaaa aagggctcca agaaggcggt gactaaggcg
     3721 cagaagaaag acggcaagaa gcgcaagcgc agccgcaagg agagctattc catctatgtg
     3781 tacaaggttc tgaagcaggt ccaccctgac accggcattt cgtccaaggc catgggcatc
     3841 atgaattcgt ttgtgaacga cattttcgag cgcatcgcag gtgaggcttc ccgcctggcg
     3901 cattacaaca agcgctcgac catcacctcc agggagatcc agacggccgt gcgcctgctg
     3961 ctgcctgggg agttggccaa gcacgccgtg tccgagggta ctaaggccgt caccaagtac
     4021 accagcgcta agggggatcc accggtcgcc accatggtga gcatggacga ggaggataac
     4081 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     4141 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     4201 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     4261 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     4321 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     4381 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     4441 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     4501 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     4561 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     4621 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     4681 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     4741 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     4801 ccctgcaggc aattcgtcga gggacctaat aacttcgtat agcatacatt atacgaagtt
     4861 atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct tatcgataat
     4921 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4981 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5041 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5101 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5161 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5221 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5281 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     5341 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5401 ccagcggacc ttccttcccg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     5461 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     5521 tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
     5581 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
     5641 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5701 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
     5761 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
     5821 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
     5881 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
     5941 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
     6001 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
     6061 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6121 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6181 gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc cagcaaaagg
     6241 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6301 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6481 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6601 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6661 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6721 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6781 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6841 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6901 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6961 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7021 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7081 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7141 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7201 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7261 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7321 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7381 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7441 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7501 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7561 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7621 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7681 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7741 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7801 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7861 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7921 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7981 aacaaatagg ggttccgcgc acatttcccc gaaaagtg
//
