LOCUS       sequence-445939-        3891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with Src kinase-dead mutant (K295R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3891)
  AUTHORS   Conway JRW, Joshi O, Kaivola J, Follain G, Gounis M, Kuhl D, Ivaska 
            J
  TITLE     Dynamic regulation of integrin beta1 phosphorylation supports 
            invasion of breast cancer cells.
  JOURNAL   Nat Cell Biol. 2025 May 26. doi: 10.1038/s41556-025-01663-4.
  PUBMED    40419795
REFERENCE   2  (bases 1 to 3891)
  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 Cell 
            Biol. 2025 May 26. doi: 10.1038/s41556-025-01663-4."
COMMENT     Sequence Label: pENTR2b-Src(K295R)
FEATURES             Location/Qualifiers
     source          1..3891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      495..504
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             501..542
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     protein_bind    complement(2120..2219)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2331..2350)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2342..3151
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2409..2428)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3241..3829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3730..3749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga
      481 ctggatccgg taccgccacc atggggagca gcaagagcaa gcccaaggac cccagccagc
      541 gccggcgcag cctggagcca cccgacagca cccaccacgg gggattccca gcctcgcaga
      601 cccccaacaa gacagcagcc cccgacacgc accgcacccc cagccgctcc tttgggaccg
      661 tggccaccga gcccaagctc ttcgggggct tcaacacttc tgacaccgtc acgtcgccgc
      721 agcgtgccgg ggcactggct ggcggcgtca ccactttcgt ggctctctat gactacgagt
      781 cccggactga aacggacttg tccttcaaga aaggagaacg cctgcagatt gtcaacaaca
      841 cggaaggtga ctggtggctg gctcattccc tcactacagg acagacgggc tacatcccca
      901 gtaactatgt cgcgccctca gactccatcc aggctgaaga gtggtacttt gggaagatca
      961 ctcgtcggga gtccgagcgg ctgctgctca accccgaaaa cccccgggga accttcttgg
     1021 tccgggagag cgagacgaca aaaggtgcct attgcctctc cgtttctgac tttgacaacg
     1081 ccaaggggct caatgtgaag cactacaaga tccgcaagct ggacagcggc ggcttctaca
     1141 tcacctcacg cacacagttc agcagcctgc agcagctggt ggcctactac tccaaacatg
     1201 ctgatggctt gtgccaccgc ctgaccaacg tctgccccac gtccaagccc cagacccagg
     1261 gactcgccaa ggacgcgtgg gaaatccccc gggagtcgct gcggctggag gtgaagctgg
     1321 ggcagggctg ctttggagag gtctggatgg ggacctggaa cggcaccacc agagtggcca
     1381 taaggactct gaagcccggc accatgtccc cggaggcctt cctgcaggaa gcccaagtga
     1441 tgaagaagct ccggcatgag aagctggttc agctgtacgc agtggtgtcg gaagagccca
     1501 tctacatcgt cactgagtac atgagcaagg ggagcctcct ggatttcctg aagggagaga
     1561 tgggcaagta cctgcggctg ccacagctcg tcgatatggc tgctcagatt gcatccggca
     1621 tggcctatgt ggagaggatg aactacgtgc accgagacct gcgggcggcc aacatcctgg
     1681 tgggggagaa cctggtgtgc aaggtggctg actttgggct ggcacgcctc atcgaggaca
     1741 acgagtacac agcacggcaa ggtgccaagt tccccatcaa gtggacagcc cccgaggcag
     1801 ccctctatgg ccggttcacc atcaagtcgg atgtctggtc cttcggcatc ctgctgactg
     1861 agctgaccac caagggccgg gtgccatacc cagggatggt caacagggag gtgctggacc
     1921 aggtggagag gggctaccgc atgccctgcc cgcccgagtg ccccgagtcg ctgcatgacc
     1981 tcatgtgcca gtgctggcgg aaggaccctg aggagcggcc cacttttgag tacctgcagg
     2041 ccttcctgga ggactacttc acctcgacag agccccagta ccagcctgga gagaacctat
     2101 agtgactcga gatatctaga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     2161 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     2221 catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2281 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2341 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2401 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2461 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2521 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2581 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2641 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2701 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2761 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2821 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2881 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2941 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3001 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3061 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3121 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta
     3181 ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3241 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3301 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3361 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3421 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3481 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3541 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3601 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3661 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3721 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3781 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3841 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
