LOCUS       sequence-485200-        4801 bp    DNA     circular SYN 03-SEP-2025
DEFINITION  Gateway entry vector with a clover-tagged phosphatase-inactive Shp2 
            mutant (D425A,C459S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4801)
  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 4801)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 3, 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-PTPN11(D425A,C459S)-Clover
FEATURES             Location/Qualifiers
     source          1..4801
                     /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"
     CDS             join(2299..2301,2302..2304,2305..3015)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2344..2365)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2952..2973
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3030..3129)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3241..3260)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3252..4061
                     /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(3319..3338)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4151..4739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4640..4659
                     /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 taccgaggag atctgccgcc gcgatcgcca tgacatcgcg gagatggttt
      541 cacccaaata tcactggtgt ggaggcagaa aacctactgt tgacaagagg agttgatggc
      601 agttttttgg caaggcctag taaaagtaac cctggagact tcacactttc cgttagaaga
      661 aatggagctg tcacccacat caagattcag aacactggtg attactatga cctgtatgga
      721 ggggagaaat ttgccacttt ggctgagttg gtccagtatt acatggaaca tcacgggcaa
      781 ttaaaagaga agaatggaga tgtcattgag cttaaatatc ctctgaactg tgcagatcct
      841 acctctgaaa ggtggtttca tggacatctc tctgggaaag aagcagagaa attattaact
      901 gaaaaaggaa aacatggtag ttttcttgta cgagagagcc agagccaccc tggagatttt
      961 gttctttctg tgcgcactgg tgatgacaaa ggggagagca atgacggcaa gtctaaagtg
     1021 acccatgtta tgattcgctg tcaggaactg aaatacgacg ttggtggagg agaacggttt
     1081 gattctttga cagatcttgt ggaacattat aagaagaatc ctatggtgga aacattgggt
     1141 acagtactac aactcaagca gccccttaac acgactcgta taaatgctgc tgaaatagaa
     1201 agcagagttc gagaactaag caaattagct gagaccacag ataaagtcaa acaaggcttt
     1261 tgggaagaat ttgagacact acaacaacag gagtgcaaac ttctctacag ccgaaaagag
     1321 ggtcaaaggc aagaaaacaa aaacaaaaat agatataaaa acatcctgcc ctttgatcat
     1381 accagggttg tcctacacga tggtgatccc aatgagcctg tttcagatta catcaatgca
     1441 aatatcatca tgcctgaatt tgaaaccaag tgcaacaatt caaagcccaa aaagagttac
     1501 attgccacac aaggctgcct gcaaaacacg gtgaatgact tttggcggat ggtgttccaa
     1561 gaaaactccc gagtgattgt catgacaacg aaagaagtgg agagaggaaa gagtaaatgt
     1621 gtcaaatact ggcctgatga gtatgctcta aaagaatatg gcgtcatgcg tgttaggaac
     1681 gtcaaagaaa gcgccgctca tgactatacg ctaagagaac ttaaactttc aaaggttgga
     1741 caagggaata cggagagaac ggtctggcaa taccactttc ggacctggcc ggcccacggc
     1801 gtgcccagcg accctggggg cgtgctggac ttcctggagg aggtgcacca taagcaggag
     1861 agcatcatgg atgcagggcc ggtcgtggtg cacagcagtg ctggaattgg ccggacaggg
     1921 acgttcattg tgattgatat tcttattgac atcatcagag agaaaggtgt tgactgcgat
     1981 attgacgttc ccaaaaccat ccagatggtg cggtctcaga ggtcagggat ggtccagaca
     2041 gaagcacagt accgatttat ctatatggcg gtccagcatt atattgaaac actacagcgc
     2101 aggattgaag aagagcagaa aagcaagagg aaagggcacg aatatacaaa tattaagtat
     2161 tctctagcgg accagacgag tggagatcag agccctctcc cgccttgtac tccaacgcca
     2221 ccctgtgcag aaatgagaga agacagtgct agagtctatg aaaacgtggg cctgatgcaa
     2281 cagcagaaaa gcggccgcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2341 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtccg cggcgagggc
     2401 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2461 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcgtggcctg cttcagccgc
     2521 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2581 caggagcgca ccatctcttt caaggacgac ggtacctaca agacccgcgc cgaggtgaag
     2641 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2701 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacg
     2761 gccgacaagc agaagaacgg catcaaggct aacttcaaga tccgccacaa cgttgaggac
     2821 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2881 ctgctgcccg acaaccacta cctgagccat cagtccgccc tgagcaaaga ccccaacgag
     2941 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggattac acatggcatg
     3001 gacgagctgt acaagtagta agaatctaga cccagctttc ttgtacaaag ttggcattat
     3061 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     3121 cattatttgc catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg
     3181 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     3241 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     3301 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     3361 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     3421 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     3481 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     3541 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     3601 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     3661 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     3721 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     3781 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     3841 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3901 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3961 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     4021 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     4081 tgtaacatta ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca
     4141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4741 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4801 t
//
