LOCUS       sequence-446288-        5363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with a clover-tagged phosphatase-inactive 
            PTP-PEST mutant (D199A,C231S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5363)
  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 5363)
  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-PTPN12(D199A,C231S)-Clover
FEATURES             Location/Qualifiers
     source          1..5363
                     /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(2860..2862,2863..2865,2866..3576)
                     /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(2905..2926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3513..3534
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3592..3691)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3803..3822)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3814..4623
                     /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(3881..3900)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4713..5301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5202..5221
                     /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 tggagcaagt ggagatcctg
      541 aggaaattca tccagagggt ccaggccatg aagagtcctg accacaatgg ggaggacaac
      601 ttcgcccggg acttcatgcg gttaagaaga ttgtctacca aatatagaac agaaaagata
      661 tatcccacag ccactggaga aaaagaagaa aatgttaaaa agaacagata caaggacata
      721 ctgccatttg atcacagccg agttaaattg acattaaaga ctccttcaca agattcagac
      781 tatatcaatg caaattttat aaagggcgtc tatgggccaa aagcatatgt agcaactcaa
      841 ggacctttag caaatacagt aatagatttt tggaggatga tatgggagta taatgttgtg
      901 atcattgtaa tggcctgccg agaatttgag atgggaagga aaaaatgtga gcgctattgg
      961 cctttgtatg gagaagaccc cataacgttt gcaccattta aaatttcttg tgaggatgaa
     1021 caagcaagaa cagactactt catcaggaca ctcttacttg aatttcaaaa tgaatctcgt
     1081 aggctgtatc agtttcatta tgtgaactgg ccagcccatg atgttccttc atcatttgat
     1141 tctattctgg acatgataag cttaatgagg aaatatcaag aacatgaaga tgttcctatt
     1201 tgtattcata gcagtgcagg ctgtggaaga acaggtgcca tttgtgccat agattatacg
     1261 tggaatttac taaaagctgg gaaaatacca gaggaattta atgtatttaa tttaatacaa
     1321 gaaatgagaa cacaaaggca ttctgcagta caaacaaagg agcaatatga acttgttcat
     1381 agagctattg cccaactgtt tgaaaaacag ctacaactat atgaaattca tggagctcag
     1441 aaaattgctg atggagtgaa tgaaattaac actgaaaaca tggtcagctc catagagcct
     1501 gaaaaacaag attctcctcc tccaaaacca ccaaggaccc gcagttgcct tgttgaaggg
     1561 gatgctaaag aagaaatact gcagccaccg gaacctcatc cagtgccacc catcttgaca
     1621 ccttctcccc cttcagcttt tccaacagtc actactgtgt ggcaggacaa tgatagatac
     1681 catccaaagc cagtgttgca tatggtttca tcagaacaac attcagcaga cctcaacaga
     1741 aactatagta aatcaacaga acttccaggg aaaaatgaat caacaattga acagatagat
     1801 aaaaaattgg aacgaaattt aagttttgag attaagaagg tccctctcca agagggacca
     1861 aaaagttttg atgggaacac acttttgaat aggggacatg caattaaaat taaatctgct
     1921 tcaccttgta tagctgataa aatctctaag ccacaggaat taagttcaga tctaaatgtc
     1981 ggtgatactt cccagaattc ttgtgtggac tgcagtgtaa cacaatcaaa caaagtttca
     2041 gttactccac cagaagaatc ccagaattca gacacacctc caaggccaga ccgcttgcct
     2101 cttgatgaga aaggacatgt aacgtggtca tttcatggac ctgaaaatgc catacccata
     2161 cctgatttat ctgaaggcaa ttcctcagat atcaactatc aaactaggaa aactgtgagt
     2221 ttaacaccaa gtcctacaac acaagttgaa acacctgatc ttgtggatca tgataacact
     2281 tcaccactct tcagaacacc cctcagtttt actaatccac ttcactctga tgactcagac
     2341 tcagatgaaa gaaactctga tggtgctgtg acccagaata aaactaatat ttcaacagca
     2401 agtgccacag tttctgctgc cactagtact gaaagcattt ctactaggaa agtattgcca
     2461 atgtccattg ctagacataa tatagcagga acaacacatt caggtgctga aaaagatgtt
     2521 gatgttagtg aagattcacc tcctccccta cctgaaagaa ctcctgaatc gtttgtgtta
     2581 gcaagtgaac ataatacacc tgtaagatcg gaatggagtg aacttcaaag tcaggaacga
     2641 tctgaacaaa aaaagtctga aggcttgata acctctgaaa atgagaaatg tgatcatcca
     2701 gcgggaggta ttcactatga aatgtgcata gaatgtccac ctactttcag tgacaagaga
     2761 gaacaaatat cagaaaatcc aacagaagcc acagatattg gttttggtaa tcgatgtgga
     2821 aaacccaaag gaccaagaga tccaccttca tgcggccgca tggtgagcaa gggcgaggag
     2881 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2941 ttcagcgtcc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
     3001 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     3061 ggcgtggcct gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3121 gccatgcccg aaggctacgt ccaggagcgc accatctctt tcaaggacga cggtacctac
     3181 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3241 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
     3301 agccacaacg tctatatcac ggccgacaag cagaagaacg gcatcaaggc taacttcaag
     3361 atccgccaca acgttgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3421 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcca tcagtccgcc
     3481 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3541 gccgggatta cacatggcat ggacgagctg tacaagtagt aagatatcta gacccagctt
     3601 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     3661 tcactatcag tcaaaataaa atcattattt gccatccagc tgcagctctg gcccgtgtct
     3721 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     3781 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     3841 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     3901 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     3961 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     4021 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     4081 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     4141 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     4201 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     4261 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     4321 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     4381 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     4441 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     4501 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     4561 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     4621 taatcagaat tggttaattg gttgtaacat tattcagatt gggccccgtt ccactgagcg
     4681 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4741 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4801 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4861 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4921 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4981 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5041 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5101 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5161 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5221 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5281 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5341 tgctggcctt ttgctcacat gtt
//
