LOCUS       sequence-373223-        7615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of shRNA targeting Cortactin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7615)
  AUTHORS   Liang X, Budnar S, Gupta S, Verma S, Han SP, Hill MM, Daly RJ, 
            Parton RG, Hamilton NA, Gomez GA, Yap AS
  TITLE     Tyrosine dephosphorylated cortactin downregulates contractility at 
            the epithelial zonula adherens through SRGAP1.
  JOURNAL   Nat Commun. 2017 Oct 5;8(1):790. doi: 10.1038/s41467-017-00797-w.
  PUBMED    28983097
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"; journalName: "Nat 
            Commun."; date: "2017-10-5"; volume: "8(1):790. doi"; pages: " 
            10.1038/s41467-017-00797-w"
COMMENT     Sequence Label: pLL5.0 hCortKD EGFP
FEATURES             Location/Qualifiers
     source          1..7615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(79..98)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        270..649
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        651..849
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     804..824
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             867..1047
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1094..1219
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1716..1949
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2134..2178
                     /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    2476..2593
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2648..2960
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2879..2899
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     3017..3033
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3075..3108)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3696..3705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3702..3704,3705..3707,3708..4421)
                     /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(3702..3704,3705..3707,3708..4418)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3747..3768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4008..4027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4355..4376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4449..4482)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4538..5124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4591..4611)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5009..5020)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5127..5143)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5653..5833
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5895..6483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5975..5994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6654..7445,7446..7514))
                     /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     7277..7296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7515..7615,1..4))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
       61 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
      121 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
      181 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
      241 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
      301 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      361 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      421 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      481 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      541 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      601 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      661 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      721 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      781 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      841 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
      901 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      961 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     1021 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     1081 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     1141 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     1201 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     1261 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1321 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1381 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1441 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1501 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1561 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     1621 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     1681 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     1741 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     1801 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     1861 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     1921 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     1981 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     2041 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     2101 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     2161 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     2221 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     2281 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     2341 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     2401 agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc
     2461 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     2521 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     2581 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt tagtaccgcg
     2641 ctctagagat ccgacgcgcc atctctaggc ccgcgccggc cccctcgcac agacttgtgg
     2701 gagaagctcg gctactcccc tgccccggtt aatttgcata taatatttcc tagtaactat
     2761 agaggcttaa tgtgcgataa aagacagata atctgttctt tttaatacta gctacatttt
     2821 acatgatagg cttggatttc tataagagat acaaatacta aattattatt ttaaaaaaca
     2881 gcacaaaagg aaactcaccc taactgtaaa gtaattgtgt gttttgagac tataaatatc
     2941 ccttggagaa aagccttgtt tgagaagcac gagtcacaga ttcaagagat ctgtgactcg
     3001 tgcttctctt ttttctcgag gtcgacggta tcgataagct cgcttcacga gattccagca
     3061 ggtcgaggga cctaataact tcgtatagca tacattatac gaagttatat taagggttcc
     3121 aagcttaagc ggccgctgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     3181 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     3241 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3301 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     3361 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3421 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3481 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     3541 acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct
     3601 tgggagggtc tcctcagatt gattgactgc ccacctcggg ggtctttcat cgaattctgc
     3661 agtcgacggt accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg
     3721 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3781 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3841 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3901 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3961 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     4021 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     4081 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4141 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4201 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4261 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4321 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4381 ccgccgggat cactctcggc atggacgagc tgtacaagta acctgcaggc aattcgtcga
     4441 gggacctaat aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg
     4501 gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     4561 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4621 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4681 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4741 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4801 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4861 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4921 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     4981 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5041 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     5101 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa
     5161 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     5221 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
     5281 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     5341 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     5401 tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt
     5461 ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga
     5521 gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     5581 gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat
     5641 ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     5701 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     5761 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     5821 aaaatctcta gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5881 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5941 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6001 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6061 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6121 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6181 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6241 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6301 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6361 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6421 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6481 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6541 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6601 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6661 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6721 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6781 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6841 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6901 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6961 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7021 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7081 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7141 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7201 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7261 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7321 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7381 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7441 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7501 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7561 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttc
//
