LOCUS       sequence-485189-        6235 bp    DNA     circular SYN 03-SEP-2025
DEFINITION  Gateway entry vector with Arg kinase-dead (K281M).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  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 6235)
  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: pENTR223-ABL2(K281M)
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1516..1527
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4114..4213)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4230..4249)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4231..4249)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4254..4270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4254..4270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4701..5492
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5585..6173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6074..6093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atggtccttg ggacagttct ccttccacct aatagttatg gcagagatca
      721 ggacacttca ctttgctgcc tgtgcactga ggcctcagaa tctgctctac ccgacttaac
      781 agaagctttg catcgtccct atggttgtga tgttgaaccc caggcactaa atgaggctat
      841 caggtggagc tccaaggaga acttgctcgg agccactgag agtgacccta atctcttcgt
      901 tgcactttat gattttgtag caagtggtga taacacactc agcatcacta aaggtgaaaa
      961 gctacgagtc cttggttaca accagaatgg tgagtggagt gaagttcgct ctaagaatgg
     1021 gcagggctgg gtgccaagca actacatcac cccagtgaac agcctggaaa aacactcctg
     1081 gtaccatgga cctgtgtcac gcagtgcagc tgagtatctg ctcagcagtc taatcaatgg
     1141 cagcttcctg gtgcgagaaa gtgagagtag ccctgggcag ctgtccatct cgctcaggta
     1201 cgagggacgt gtgtatcact acaggatcaa taccactgca gatggcaagg tgtatgtgac
     1261 tgctgagagc cgcttcagca ccttggcaga gcttgtacac catcactcca cagtggctga
     1321 tgggctggtg acaacattac actacccagc acccaagtgt aataagccta cagtctatgg
     1381 tgtgtccccc atccacgaca aatgggaaat ggagcgaaca gatattacca tgaagcacaa
     1441 acttgggggc ggtcagtatg gagaggttta cgttggcgtc tggaagaaat acagccttac
     1501 agttgctgtg atgacattga aggaagatac catggaggta gaagaattcc tgaaagaagc
     1561 tgcagtaatg aaggaaatca agcatcctaa tctggtacaa cttttaggtg tgtgtacttt
     1621 ggagccacca ttttacattg tgactgaata catgccatac gggaatttgc tggattacct
     1681 ccgagaatgc aaccgagaag aggtgactgc agttgtgctg ctctacatgg ccactcagat
     1741 ttcttctgca atggagtact tagagaagaa gaatttcatc catagagatc ttgcagctcg
     1801 taactgccta gtgggagaaa accatgtggt aaaagtggct gactttggct taagtagatt
     1861 gatgactgga gacacttata ctgctcatgc tggagccaaa tttcctatta agtggacagc
     1921 accagagagt cttgcctaca ataccttctc aattaaatct gacgtctggg cttttggggt
     1981 attgttgtgg gaaattgcta cctatggaat gtcaccatat ccaggtattg acctgtctca
     2041 ggtctatgac ctactagaaa aaggatatcg aatggaacag cctgagggat gcccccctaa
     2101 ggtttatgaa cttatgagag catgctggaa gtggagccct gccgataggc cctcttttgc
     2161 tgaaacacac caagcttttg aaaccatgtt ccatgactcc agcatttctg aagaggtagc
     2221 tgaggagctt gggagagccg cctcctcgtc atctgttgtt ccatacctgc cccggctacc
     2281 tatacttcct tccaagactc ggacactgaa gaaacaggtg gagaacaagg agaacattga
     2341 aggggcacaa gatgccacag aaaattctgc ttccagttta gcaccagggt tcatcagagg
     2401 tgcacaggcc tctagtggat ccccagcact gcctcgaaag caaagagaca agtcacccag
     2461 cagcctcttg gaagatgcca aagagacatg cttcaccagg gataggaagg ggggcttctt
     2521 cagctccttc atgaagaaga gaaatgctcc cacacccccc aaacgcagca gctccttccg
     2581 agaaatggag aatcagcccc ataagaaata cgaactcacg ggtaacttct catctgttgc
     2641 ttctctacag catgctgatg ggttctcttt cactcctgcc cagcaagagg cgaatctggt
     2701 gccacccaag tgctatgggg ggagctttgc acagaggaac ctctgtaatg acgacggtgg
     2761 tgggggtggg ggcagtggca ctgctggggg tgggtggtct ggcatcacag gcttctttac
     2821 accacgctta atcaaaaaga cactgggctt acgagcaggt aaacccacag ccagtgatga
     2881 cacttccaag ccttttccaa ggtcaaactc tacatcttcc atgtcctcag ggcttccaga
     2941 gcaggatagg atggcaatga cccttcccag gaactgccag aggtccaaac tccagctgga
     3001 aaggacagtg tccacctctt ctcagccaga agagaatgtg gacagggcca atgacatgct
     3061 tccaaaaaaa tcagaggaaa gtgctgctcc aagcagggag agaccaaaag ccaagttatt
     3121 gcccagagga gccacagctc ttcctctcag aacaccctct ggggatctag ccattacaga
     3181 gaaggaccct ccaggggtgg gagtggctgg agtggcagct gcccccaagg gtaaagagaa
     3241 gaatggtggg gcacgacttg ggatggctgg agttccagag gatggagagc agccgggctg
     3301 gccttctcca gccaaggctg cccccgtcct cccaaccact cacaaccaca aagtgccagt
     3361 ccttatctca cccactctga aacacactcc agctgacgtg cagctcattg gcacagactc
     3421 tcaggggaat aaattcaagc tcttatctga gcatcaggtc acatcctctg gagacaagga
     3481 ccgaccccga cgggtaaaac caaagtgtgc cccaccccca ccaccagtga tgagactact
     3541 gcagcatccg tccatctgct cagaccctac agaagagcca actgccctaa ctgcaggaca
     3601 gtccacatca gaaacacagg aaggaggaaa gaaggcagct ctgggcgcag tgcccatcag
     3661 tgggaaagct gggaggccag tgatgcctcc acctcaagtg cctctgccca catcttccat
     3721 ctcgccagcc aaaatggcca atggcacagc aggtactaaa gtggctctga gaaaaaccaa
     3781 acaggccgct gagaaaatct cagcagacaa aatcagcaaa gaggccctgc tggaatgtgc
     3841 tgacctactg tccagtgcac tcacggaacc tgtgcccaac agccagctgg tagacactgg
     3901 acaccagctg cttgactact gctcaggcta tgtggactgc atccctcaaa ctcgcaacaa
     3961 atttgccttc cgagaggctg tgagcaaact ggaactcagc ctgcaggagc tacaggtttc
     4021 ttcagcagct gctggtgtgc ccgggacaaa ccctgtcctt aataacttat tgtcatgtgt
     4081 acaggaaatc agtgatgtgg tgcagaggta gacacccagc tttcttgtac aaagttggca
     4141 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     4201 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     4261 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     4321 aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     4381 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     4441 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     4501 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     4561 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     4621 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     4681 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     4741 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     4801 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     4861 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     4921 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     4981 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     5041 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     5101 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     5161 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     5221 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     5281 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     5341 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     5401 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     5461 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     5521 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5581 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5641 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5701 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5761 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5821 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5881 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5941 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6001 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6061 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6121 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6181 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
