LOCUS       sequence-373219-        9271 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral expression of shRNA targeting Cortactin + reconstitution 
            of Cortactin3YF 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 Apr 14, 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-Flcortmut3YF mCherry
FEATURES             Location/Qualifiers
     source          1..9271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(67..86)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        258..637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     792..812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             855..1035
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1082..1207
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1704..1937
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2122..2166
                     /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    2464..2581
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2636..2948
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2867..2887
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     3005..3021
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3063..3096)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      5312..5321
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5463..5481)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5731..5750
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(6093..6126)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6182..6768
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6235..6255)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6653..6664)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6771..6787)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7297..7477
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7539..8127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7619..7638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8298..9089,9090..9158))
                     /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     8921..8940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9159..9263)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
       61 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
      121 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
      181 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
      241 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
      301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      601 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc
      841 gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
      901 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
      961 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     1021 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     1081 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     1141 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     1201 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     1261 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     1321 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     1381 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     1441 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     1501 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc
     1561 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     1621 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     1681 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     1741 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     1801 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     1861 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     1921 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     1981 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     2041 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     2101 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     2161 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     2221 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     2281 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     2341 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     2401 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
     2461 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     2521 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     2581 tcgggtttat tacagggaca gcagagatcc agtttggtta gtaccgcgct ctagagatcc
     2641 gacgcgccat ctctaggccc gcgccggccc cctcgcacag acttgtggga gaagctcggc
     2701 tactcccctg ccccggttaa tttgcatata atatttccta gtaactatag aggcttaatg
     2761 tgcgataaaa gacagataat ctgttctttt taatactagc tacattttac atgataggct
     2821 tggatttcta taagagatac aaatactaaa ttattatttt aaaaaacagc acaaaaggaa
     2881 actcacccta actgtaaagt aattgtgtgt tttgagacta taaatatccc ttggagaaaa
     2941 gccttgtttg agaagcacga gtcacagatt caagagatct gtgactcgtg cttctctttt
     3001 ttctcgaggt cgacggtatc gataagctcg cttcacgaga ttccagcagg tcgagggacc
     3061 taataacttc gtatagcata cattatacga agttatatta agggttccaa gcttaagcgg
     3121 ccgctgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     3181 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     3241 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3301 aagaacagat ggtccccaga tgcggtcgcc ctcagcagtt tctagagaac catcagatgt
     3361 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     3421 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     3481 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
     3541 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
     3601 cagattgatt gactgcccac ctcgggggtc tttcatcgaa ttcgccacca tgtggaaagc
     3661 ttcagcaggc cacgctgtgt ccatcgccca ggatgacgcg ggggccgatg actgggagac
     3721 tgaccctgat tttgtgaatg atgtgagtga gaaggagcaa agatggggtg ccaagacggt
     3781 gcagggctcc gggcaccagg agcatatcaa catacacaag ctgagggaga atgtctttca
     3841 agagcatcag acccttaagg agaaggaact tgaaacagga ccaaaagctt cccatggcta
     3901 tggagggaaa tttggtgtgg aacaagaccg aatggataag tcagctgtcg gccacgaata
     3961 tcagtcgaaa ctttccaagc actgctcgca ggtggactcg gtccgtggct tcggaggcaa
     4021 gtttggtgtc cagatggaca gagttgatca gtctgctgta ggctttgaat accaggggaa
     4081 gactgagaag catgcctccc agaaagacta ctccagtggt tttggcggca agtatggcgt
     4141 gcaggccgac cgagtagaca agagcgcggt gggcttcgac taccagggca agacggagaa
     4201 acatgaatcc cagagagatt actccaaagg tttcggcggc aaatacggta tcgacaagga
     4261 caaagtggat aagagcgccg ttggctttga gtatcaaggc aaaacggaga agcacgagtc
     4321 ccagaaagac tatgtgaaag ggtttggagg aaaatttggt gtgcagacag acagacaaga
     4381 caaatgtgcc cttggctggg atcaccagga gaaattgcag ctgcatgaat cccaaaaaga
     4441 ttataagact ggttttggag gcaaattcgg tgttcagtcg gagaggcagg actccgctgc
     4501 tgtggggttt gattacaagg agaagctggc caagcacgag tcccagcaag actactccaa
     4561 aggattcggc gggaagtatg gggtgcagaa ggatcggatg gataagaatg cgtcaacctt
     4621 tgaggatgtc acccaggtgt cctctgccta ccagaagaca gtacctgtcg aagctgtgac
     4681 cagcaaaaca agtaacatca gagctaactt tgaaaacctc gctaaggaga aagagcagga
     4741 ggacaggcgg aaggcggagg cggagagagc ccagcggatg gccaaggagc ggcaggagca
     4801 ggaagaggcc aggaggaagc tggaggagca agccagagcc aaaacgcaaa cgccccctgt
     4861 gtcgcccgca cctcagccaa ccgaggagag gctgccctcg agccccgtct ttgaggatgc
     4921 ggcttccttc aaggcagagc tgagctacag aggccctgtg agtgggacgg agccggagcc
     4981 cgtgtacagc atggaggccg ctgactaccg agaggccagc agccagcagg gcctggccta
     5041 tgccacagag gctgtctttg aaagcgcaga ggccccgggc cactatcccg cagaggacag
     5101 cacctttgat gagtacgaga acgatctggg gatcacagcc gtcgccctgt acgactacca
     5161 ggctgcgggc gatgatgaga tctcatttga ccctgatgac atcatcacca acatcgagat
     5221 gattgacgac ggctggtggc gcggggtgtg caagggccgg tacgggctct tcccagccaa
     5281 ctatgtggag ctgcggcagg atccaccggt cgccaccatg gtgagcatgg acgaggagga
     5341 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     5401 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     5461 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     5521 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     5581 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     5641 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     5701 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     5761 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     5821 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     5881 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     5941 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     6001 cggcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     6061 accaccctgc aggcaattcg tcgagggacc taataacttc gtatagcata cattatacga
     6121 agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga
     6181 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6241 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6301 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6361 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6421 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6481 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6541 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     6601 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6661 caatccagcg gaccttcctt cccggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6721 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     6781 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     6841 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     6901 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6961 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     7021 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     7081 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     7141 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     7201 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     7261 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     7321 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7381 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7441 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa
     7501 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7561 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7621 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7681 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7741 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7801 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7861 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7921 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7981 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8041 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8101 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8161 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8221 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8281 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8341 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8401 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8461 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8521 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8581 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8641 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8701 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8761 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8821 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8881 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8941 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9001 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9061 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9121 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9181 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9241 aataaacaaa taggggttcc gcgcacattt c
//
