LOCUS       sequence-373222-        9271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of shRNA targeting Cortactin + reconstitution 
            of Cortactin3YD mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9271)
  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 9271)
  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 mCherry-hCortKD-Flcortmut3YD
FEATURES             Location/Qualifiers
     source          1..9271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..49)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        221..600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        602..800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     755..775
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             818..998
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1045..1170
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1667..1900
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2085..2129
                     /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    2427..2544
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2599..2911
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2830..2850
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2968..2984
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3026..3059)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      5275..5284
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5426..5444)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5694..5713
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(6056..6089)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6145..6731
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6198..6218)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6616..6627)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6734..6750)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7260..7440
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7502..8090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7582..7601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8261..9052,9053..9121))
                     /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     8884..8903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9122..9226)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
       61 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
      121 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      181 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      241 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      301 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      361 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      421 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      481 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      541 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      601 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      661 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      721 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      781 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
      841 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      901 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
      961 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     1021 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     1081 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     1141 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     1201 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     1261 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     1321 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     1381 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     1441 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     1501 cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1561 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1621 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1681 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1741 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1801 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1861 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1921 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1981 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2041 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2101 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2161 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2221 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2281 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2341 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2401 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2461 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2521 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2581 ttagtaccgc gctctagaga tccgacgcgc catctctagg cccgcgccgg ccccctcgca
     2641 cagacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat ataatatttc
     2701 ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct ttttaatact
     2761 agctacattt tacatgatag gcttggattt ctataagaga tacaaatact aaattattat
     2821 tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg tgttttgaga
     2881 ctataaatat cccttggaga aaagccttgt ttgagaagca cgagtcacag attcaagaga
     2941 tctgtgactc gtgcttctct tttttctcga ggtcgacggt atcgataagc tcgcttcacg
     3001 agattccagc aggtcgaggg acctaataac ttcgtatagc atacattata cgaagttata
     3061 ttaagggttc caagcttaag cggccgctga aagaccccac ctgtaggttt ggcaagctag
     3121 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     3181 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     3241 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc gccctcagca
     3301 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     3361 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     3421 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     3481 cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc
     3541 gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatc
     3601 gaattcgcca ccatgtggaa agcttcagca ggccacgctg tgtccatcgc ccaggatgac
     3661 gcgggggccg atgactggga gactgaccct gattttgtga atgatgtgag tgagaaggag
     3721 caaagatggg gtgccaagac ggtgcagggc tccgggcacc aggagcatat caacatacac
     3781 aagctgaggg agaatgtctt tcaagagcat cagaccctta aggagaagga acttgaaaca
     3841 ggaccaaaag cttcccatgg ctatggaggg aaatttggtg tggaacaaga ccgaatggat
     3901 aagtcagctg tcggccacga atatcagtcg aaactttcca agcactgctc gcaggtggac
     3961 tcggtccgtg gcttcggagg caagtttggt gtccagatgg acagagttga tcagtctgct
     4021 gtaggctttg aataccaggg gaagactgag aagcatgcct cccagaaaga ctactccagt
     4081 ggttttggcg gcaagtatgg cgtgcaggcc gaccgagtag acaagagcgc ggtgggcttc
     4141 gactaccagg gcaagacgga gaaacatgaa tcccagagag attactccaa aggtttcggc
     4201 ggcaaatacg gtatcgacaa ggacaaagtg gataagagcg ccgttggctt tgagtatcaa
     4261 ggcaaaacgg agaagcacga gtcccagaaa gactatgtga aagggtttgg aggaaaattt
     4321 ggtgtgcaga cagacagaca agacaaatgt gcccttggct gggatcacca ggagaaattg
     4381 cagctgcatg aatcccaaaa agattataag actggttttg gaggcaaatt cggtgttcag
     4441 tcggagaggc aggactccgc tgctgtgggg tttgattaca aggagaagct ggccaagcac
     4501 gagtcccagc aagactactc caaaggattc ggcgggaagt atggggtgca gaaggatcgg
     4561 atggataaga atgcgtcaac ctttgaggat gtcacccagg tgtcctctgc ctaccagaag
     4621 acagtacctg tcgaagctgt gaccagcaaa acaagtaaca tcagagctaa ctttgaaaac
     4681 ctcgctaagg agaaagagca ggaggacagg cggaaggcgg aggcggagag agcccagcgg
     4741 atggccaagg agcggcagga gcaggaagag gccaggagga agctggagga gcaagccaga
     4801 gccaaaacgc aaacgccccc tgtgtcgccc gcacctcagc caaccgagga gaggctgccc
     4861 tcgagccccg tcgatgagga tgcggcttcc ttcaaggcag agctgagcta cagaggccct
     4921 gtgagtggga cggagccgga gcccgtgtac agcatggagg ccgctgacta ccgagaggcc
     4981 agcagccagc agggcctggc ctatgccaca gaggctgtcg atgaaagcgc agaggccccg
     5041 ggccactatc ccgcagagga cagcaccgac gatgagtacg agaacgatct ggggatcaca
     5101 gccgtcgccc tgtacgacta ccaggctgcg ggcgatgatg agatctcatt tgaccctgat
     5161 gacatcatca ccaacatcga gatgattgac gacggctggt ggcgcggggt gtgcaagggc
     5221 cggtacgggc tcttcccagc caactatgtg gagctgcggc aggatccacc ggtcgccacc
     5281 atggtgagca tggacgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     5341 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     5401 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     5461 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     5521 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     5581 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     5641 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     5701 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     5761 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     5821 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     5881 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     5941 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta aagcggccgc
     6001 gactctagat cataatcagc cataccaccc tgcaggcaat tcgtcgaggg acctaataac
     6061 ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt ccactaggta
     6121 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     6181 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     6241 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     6301 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     6361 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6421 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6481 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     6541 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6601 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccggcc tgctgccggc
     6661 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     6721 cgcctccccg catcgatacc gtcgacctcg atcgagacct agaaaaacat ggagcaatca
     6781 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg
     6841 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     6901 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6961 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     7021 ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca
     7081 agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct
     7141 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga
     7201 ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg
     7261 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7321 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7381 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7441 gcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7681 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7921 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     8041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9241 aagtgccacc tgacgtcgac ggatcgggag a
//
