LOCUS       sequence-442159-        8885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This retroviral plasmid expresses a mutated cDNA (dominant negative:
            Y861F) of human PTK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8885)
  AUTHORS   Pozzato C, Outeiro-Pinho G, Galie M, Ramadori G, Konstantinidou G
  TITLE     ERK5 suppression overcomes FAK inhibitor resistance in mutant 
            KRAS-driven non-small cell lung cancer.
  JOURNAL   EMBO Mol Med. 2024 Sep 13. doi: 10.1038/s44321-024-00138-7.
  PUBMED    39271958
REFERENCE   2  (bases 1 to 8885)
  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: "EMBO Mol 
            Med. 2024 Sep 13. doi: 10.1038/s44321-024-00138-7."
COMMENT     Sequence Label: pWZL-hygro-FAKY861F
FEATURES             Location/Qualifiers
     source          1..8885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             21..490
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    555..754
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             955..1371
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1301..1323
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1339..1355
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4712..5173
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4768..4785)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4995..5014
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5178..6203
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5178..6203
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             6392..6861
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6991..7008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7162..7750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7242..7261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7910..8701,8702..8770))
                     /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     8533..8552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8771..8875)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgaaaagtgc cacctgcagc ctgaatatgg gccaaacagg atatctgtgg taagcagttc
       61 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
      121 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
      181 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
      241 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
      301 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
      361 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
      421 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
      481 gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc
      541 caccaccggg aggcaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct
      601 atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg tatctggcgg
      661 acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg
      721 gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc
      781 ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct
      841 gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc
      901 atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag
      961 actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct
     1021 cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg
     1081 ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag
     1141 gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc
     1201 gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct
     1261 aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg
     1321 accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggccggatc
     1381 ccagtgtggt ggtacgtagg aattcgccag cacagtggtc gaagcggatc catgatctcg
     1441 gctgactgca acctctgcct cccagaatat gacagatacc tagcatctag caaaataatg
     1501 gcagctgctt accttgaccc caacttgaat cacacaccaa attcgagtac taagactcac
     1561 ctgggtactg gtatggaacg ttctcctggt gcaatggagc gagtattaaa ggtctttcat
     1621 tattttgaaa gcaatagtga gccaaccacc tgggccagta ttatcaggca tggagatgct
     1681 actgatgtca ggggcatcat tcagaagata gtggacagtc acaaagtaaa gcatgtggcc
     1741 tgctatggat tccgcctcag tcacctgcgg tcagaggagg ttcactggct tcacgtggat
     1801 atgggcgtct ccagtgtgag ggagaagtat gagcttgctc acccaccaga ggagtggaaa
     1861 tatgaattga gaattcgtta tttgccaaaa ggatttctaa accagtttac tgaagataag
     1921 ccaactttga atttcttcta tcaacaggtg aagagcgatt atatgttaga gatagctgat
     1981 caagtggacc aggaaattgc tttgaagttg ggttgtctag aaatacggcg atcatactgg
     2041 gagatgcggg gcaatgcact agaaaagaag tctaactatg aagtattaga aaaagatgtt
     2101 ggtttaaagc gattttttcc taagagttta ctggattctg tcaaggccaa aacactaaga
     2161 aaactgatcc aacaaacatt tagacaattt gccaacctta atagagaaga aagtattctg
     2221 aaattctttg agatcctgtc tccagtctac agatttgata aggaatgctt caagtgtgct
     2281 cttggttcaa gctggattat ttcagtggaa ctggcaatcg gcccagaaga aggaatcagt
     2341 tacctaacgg acaagggctg caatcccaca catcttgctg acttcactca agtgcaaacc
     2401 attcagtatt caaacagtga agacaaggac agaaaaggaa tgctacaact aaaaatagca
     2461 ggtgcacccg agcctctgac agtgacggca ccatccctaa ccattgcgga gaatatggct
     2521 gacctaatag atgggtactg ccggctggtg aatggaacct cgcagtcatt tatcatcaga
     2581 cctcagaaag aaggtgaacg ggctttgcca tcaataccaa agttggccaa cagcgaaaag
     2641 caaggcatgc ggacacacgc cgtctctgtg tcagaaacag atgattatgc tgagattata
     2701 gatgaagaag atacttacac catgccctca accagggatt atgagattca aagagaaaga
     2761 atagaacttg gacgatgtat tggagaaggc caatttggag atgtacatca aggcatttat
     2821 atgagtccag agaatccagc tttggcggtt gcaattaaaa catgtaaaaa ctgtacttcg
     2881 gacagcgtga gagagaaatt tcttcaagaa gccttaacaa tgcgtcagtt tgaccatcct
     2941 catattgtga agctgattgg agtcatcaca gagaatcctg tctggataat catggagctg
     3001 tgcacacttg gagagctgag gtcatttttg caagtaagga aatacagttt ggatctagca
     3061 tctttgatcc tgtatgccta tcagcttagt acagctcttg catatctaga gagcaaaaga
     3121 tttgtacaca gggacattgc tgctcggaat gttctggtgt cctcaaatga ttgtgtaaaa
     3181 ttaggagact ttggattatc ccgatatatg gaagatagta cttactacaa agcttccaaa
     3241 ggaaaattgc ctattaaatg gatggctcca gagtcaatca attttcgacg ttttacctca
     3301 gctagtgacg tatggatgtt tggtgtgtgt atgtgggaga tactgatgca tggtgtgaag
     3361 ccttttcaag gagtgaagaa caatgatgta atcggtcgaa ttgaaaatgg ggaaagatta
     3421 ccaatgcctc caaattgtcc tcctaccctc tacagcctta tgacgaaatg ctgggcctat
     3481 gaccccagca ggcggcccag gtttactgaa cttaaagctc agctcagcac aatcctggag
     3541 gaagagaagg ctcagcaaga agagcgcatg aggatggagt ccagaagaca ggccacagtg
     3601 tcctgggact ccggagggtc tgatgaagca ccgcccaagc ccagcagacc gggttatccc
     3661 agtccgaggt ccagcgaagg attttatccc agcccacagc acatggtaca aaccaatcat
     3721 taccaggttt ctggctaccc tggttcacat ggaatcacag ccatggctgg cagcatctat
     3781 ccaggtcagg catctctttt ggaccaaaca gattcatgga atcatagacc tcaggagata
     3841 gcaatgtggc agcccaatgt ggaggactct acagtattgg acctgcgagg gattgggcaa
     3901 gtgttgccaa cccatctgat ggaagagcgt ctaatccgac agcaacagga aatggaagaa
     3961 gatcagcgct ggctggaaaa agaggaaaga tttctgaaac ctgatgtgag actctctcga
     4021 ggcagtattg acagggagga tggaagtctt cagggtccga ttggaaacca acatatattt
     4081 cagcctgtgg gtaaaccaga tcctgcagct ccaccaaaga aaccgcctcg ccctggagct
     4141 cccggtcatc tgggaagcct tgccagcctc agcagccctg ctgacagcta caacgagggt
     4201 gtcaagcttc agccccagga aatcagcccc cctcctactg ccaacctgga ccggtcgaat
     4261 gataaggtgt acgagaatgt gacgggcctg gtgaaagctg tcatcgagat gtccagtaaa
     4321 atccagccag ccccaccaga ggagtatgtc cctatggtga aggaagtcgg cttggccctg
     4381 aggacattat tggccactgt ggatgagacc attcccctcc taccagccag cacccaccga
     4441 gagattgaga tggcacagaa gctattgaac tctgacctgg gtgagctcat caacaagatg
     4501 aaactggccc agcagtatgt catgaccagc ctccagcaag agtacaaaaa gcaaatgctg
     4561 actgctgctc acgccctggc tgtggatgcc aaaaacttac tcgatgtcat tgaccaagca
     4621 agactgaaaa tgcttgggca gacgagacca cactgagtcg actcgaccaa ttccggttat
     4681 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     4741 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     4801 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     4861 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     4921 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     4981 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     5041 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     5101 agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     5161 aacacgatga taataccatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     5221 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     5281 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     5341 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     5401 cggaagtgct tgacattggg gaatttagcg agagcctgac ctattgcatc tcccgccgtg
     5461 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     5521 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     5581 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     5641 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     5701 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     5761 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     5821 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     5881 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     5941 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     6001 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     6061 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     6121 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     6181 ctcgtccgag ggcaaaggaa tagagtagat gccgaccggg atctatcgat aaaataaaag
     6241 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     6301 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     6361 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     6421 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     6481 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     6541 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     6601 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     6661 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     6721 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     6781 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     6841 cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt
     6901 aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg
     6961 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7021 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7081 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7141 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7201 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7261 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7321 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7381 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7441 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7501 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7561 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7621 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7681 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7741 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7801 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7861 gcggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7921 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7981 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8041 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8101 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8161 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8221 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8281 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8341 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8401 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8461 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8521 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8581 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8641 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8701 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8761 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8821 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8881 ttccc
//
