LOCUS       sequence-446281-        4742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with EGFP-tagged paxillin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4742)
  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 4742)
  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-Paxillin-EGFP
FEATURES             Location/Qualifiers
     source          1..4742
                     /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      509..518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2231..2240
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2237..2239,2240..2242,2243..2956)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2237..2239,2240..2242,2243..2953)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2282..2303)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2543..2562)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2890..2911
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2971..3070)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3182..3201)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3193..4002
                     /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(3260..3279)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4092..4680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4581..4600
                     /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 taccgaattc gcggccgcgc caccatggac gacctcgacg ccctgctggc
      541 ggacttggag tctaccacct cccacatctc caaacggcct gtgttcttgt cggaggagac
      601 cccctactca tacccaactg gaaaccacac ataccaggag attgccgtgc caccccccgt
      661 ccccccaccc ccgtccagcg aggccctcaa tggcacaatc cttgacccct tagaccagtg
      721 gcagcccagc ggctcccgat tcatccacca gcagcctcag tcctcatcac ctgtgtacgg
      781 ctccagtgcc aaaacttcca gtgtctccaa ccctcaggac agtgttggct ctccgtgctc
      841 ccgagtgggt gaggaggagc acgtctacag cttccccaac aagcagaaat cagctgagcc
      901 ttcacccacc gtaatgagca cgtccctggg cagcaacctt tctgaactcg accgcctgct
      961 gctggaactg aacgctgtac agcataaccc gccaggcttc cctgcagatg aggccaactc
     1021 aagccccccg cttcctgggg ccctgagccc cctctatggt gtcccagaga ctaacagccc
     1081 cttgggaggc aaagctgggc ccctgacgaa agagaagcct aagcggaatg ggggccgggg
     1141 cctggaggac gtgcggccca gtgtggagag tctcttggat gaactggaga gctccgtgcc
     1201 cagccccgtc cctgccatca ctgtgaacca gggcgagatg agcagcccgc agcgcgtcac
     1261 ctccacccaa cagcagacac gcatctcggc ctcctctgcc accagggagc tggacgagct
     1321 gatggcttcg ctgtcggatt tcaagttcat ggcccagggg aagacaggga gcagctcacc
     1381 ccctgggggg cccccgaagc ccgggagcca gctggacagc atgctgggga gcctgcagtc
     1441 tgacctgaac aagctggggg tcgccacagt cgccaaagga gtctgcgggg cctgcaagaa
     1501 gcccatcgcc gggcaggttg tgaccgccat ggggaagacg tggcaccccg agcacttcgt
     1561 ctgcacccac tgccaggagg agatcggatc ccggaacttc ttcgagcggg atggacagcc
     1621 ctactgtgaa aaggactacc acaacctctt ctccccgcgc tgctactact gcaacggccc
     1681 catcctggat aaagtggtga cagcccttga ccggacgtgg caccctgaac acttcttctg
     1741 tgcacagtgt ggagccttct ttggtcccga agggttccac gagaaggacg gcaaggccta
     1801 ctgtcgcaag gactacttcg acatgttcgc acccaagtgt ggcggctgcg cccgggccat
     1861 cctggagaac tatatctcag ccctcaacac gctgtggcat cctgagtgct ttgtgtgccg
     1921 ggaatgcttc acgccattcg tgaacggcag cttcttcgag cacgacgggc agccctactg
     1981 tgaggtgcac taccacgagc ggcgcggctc gctgtgttct ggctgccaga agcccatcac
     2041 cggccgctgc atcaccgcca tggccaagaa gttccacccc gagcacttcg tctgtgcctt
     2101 ctgcctcaag cagctcaaca agggcacctt caaggagcag aacgacaagc cttactgtca
     2161 gaactgcttc ctcaagctct tctgccgaat tctgcagtcg acggtaccgc gggcccggga
     2221 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2281 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2341 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2401 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2461 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2521 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2581 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2641 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2701 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2761 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2821 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2881 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2941 cgagctgtac aagtaactcg agatatctag acccagcttt cttgtacaaa gttggcatta
     3001 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     3061 tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     3121 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     3181 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     3241 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     3301 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     3361 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     3421 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     3481 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     3541 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     3601 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     3661 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3721 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3781 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3841 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3901 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3961 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     4021 ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt agaaaagatc
     4081 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4141 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4201 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4261 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4321 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4381 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4441 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4501 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4561 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4621 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4681 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4741 tt
//
