LOCUS       sequence-445868-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with integrin beta1 NPxY mutant (YYFF) tagged 
            with mRuby2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  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 5412)
  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-ITGB1(YYFF)-mRuby2
FEATURES             Location/Qualifiers
     source          1..5412
                     /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             2910..3620
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     protein_bind    complement(3641..3740)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3852..3871)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3863..4672
                     /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(3930..3949)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4762..5350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5251..5270
                     /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 taccaccgcc atgaatttac aaccaatttt ctggattgga ctgatcagtt
      541 cagtttgctg tgtgtttgct caaacagatg aaaatagatg tttaaaagca aatgccaaat
      601 catgtggaga atgtatacaa gcagggccaa attgtgggtg gtgcacaaat tcaacatttt
      661 tacaggaagg aatgcctact tctgcacgat gtgatgattt agaagcctta aaaaagaagg
      721 gttgccctcc agatgacata gaaaatccca gaggctccaa agatataaag aaaaataaaa
      781 atgtaaccaa ccgtagcaaa ggaacagcag agaagctcaa gccagaggat attactcaga
      841 tccaaccaca gcagttggtt ttgcgattaa gatcagggga gccacagaca tttacattaa
      901 aattcaagag agctgaagac tatcccattg acctctacta ccttatggac ctgtcttact
      961 caatgaaaga cgatttggag aatgtaaaaa gtcttggaac agatctgatg aatgaaatga
     1021 ggaggattac ttcggacttc agaattggat ttggctcatt tgtggaaaag actgtgatgc
     1081 cttacattag cacaacacca gctaagctca ggaacccttg cacaagtgaa cagaactgca
     1141 ccagcccatt tagctacaaa aatgtgctca gtcttactaa taaaggagaa gtatttaatg
     1201 aacttgttgg aaaacagcgc atatctggaa atttggattc tccagaaggt ggtttcgatg
     1261 ccatcatgca agttgcagtt tgtggatcac tgattggctg gaggaatgtt acacggctgc
     1321 tggtgttttc cacagatgcc gggtttcact ttgctggaga tgggaaactt ggtggcattg
     1381 ttttaccaaa tgatggacaa tgtcacctgg aaaataatat gtacacaatg agccattatt
     1441 atgattatcc ttctattgct caccttgtcc agaaactgag tgaaaataat attcagacaa
     1501 tttttgcagt tactgaagaa tttcagcctg tttacaagga gctgaaaaac ttgatcccta
     1561 agtcagcagt aggaacatta tctgcaaatt ctagcaatgt aattcagttg atcattgatg
     1621 catacaattc cctttcctca gaagtcattt tggaaaacgg caaattgtca gaaggagtaa
     1681 caataagtta caaatcttac tgcaagaacg gggtgaatgg aacaggggaa aatggaagaa
     1741 aatgttccaa tatttccatt ggagatgagg ttcaatttga aattagcata acttcaaata
     1801 agtgtccaaa aaaggattct gacagcttta aaattaggcc tctgggcttt acggaggaag
     1861 tagaggttat tcttcagtac atctgtgaat gtgaatgcca aagcgaaggc atccctgaaa
     1921 gtcccaagtg tcatgaagga aatgggacat ttgagtgtgg cgcgtgcagg tgcaatgaag
     1981 ggcgtgttgg tagacattgt gaatgcagca cagatgaagt taacagtgaa gacatggatg
     2041 cttactgcag gaaagaaaac agttcagaaa tctgcagtaa caatggagag tgcgtctgcg
     2101 gacagtgtgt ttgtaggaag agggataata caaatgaaat ttattctggc aaattctgcg
     2161 agtgtgataa tttcaactgt gatagatcca atggcttaat ttgtggagga aatggtgttt
     2221 gcaagtgtcg tgtgtgtgag tgcaacccca actacactgg cagtgcatgt gactgttctt
     2281 tggatactag tacttgtgaa gccagcaacg gacagatctg caatggccgg ggcatctgcg
     2341 agtgtggtgt ctgtaagtgt acagatccga agtttcaagg gcaaacgtgt gagatgtgtc
     2401 agacctgcct tggtgtctgt gctgagcata aagaatgtgt tcagtgcaga gccttcaata
     2461 aaggagaaaa gaaagacaca tgcacacagg aatgttccta ttttaacatt accaaggtag
     2521 aaagtcggga caaattaccc cagccggtcc aacctgatcc tgtgtcccat tgtaaggaga
     2581 aggatgttga cgactgttgg ttctatttta cgtattcagt gaatgggaac aacgaggtca
     2641 tggttcatgt tgtggagaat ccagagtgtc ccactggtcc agacatcatt ccaattgtag
     2701 ctggtgtggt tgctggaatt gttcttattg gccttgcgtt actcctcatt tggaagcttt
     2761 taatgataat tcatgacaga agggagtttg ctaaatttga aaaggagaaa atgaatgcca
     2821 aatgggacac gggtgaaaat cctattttta agagtgccgt aacaactgtg gtcaatccga
     2881 agtttgaggg aaaaggcggc ggcggccgca tggtgtctaa gggcgaagag ctgatcaagg
     2941 aaaatatgcg tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca
     3001 caggtgaagg agaaggcaat ccgtacatgg gaactcaaac catgaggatc aaagtcatcg
     3061 agggaggacc cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc
     3121 gtacttttat caagtacccg aaaggcattc ctgatttctt taaacagtcc tttcctgagg
     3181 gttttacttg ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcatgcagg
     3241 acaccagcct tgaggatggc tgtctcgttt accacgtcca agtcagaggg gtaaactttc
     3301 cctccaatgg tcccgtgatg cagaagaaga ccaagggttg ggagcctaat acagagatga
     3361 tgtatccagc agatggtggt ctgaggggat acactcatat ggcactgaaa gttgatggtg
     3421 gtggccatct gtcttgctct ttcgtaacaa cttacaggtc aaaaaagacc gtcgggaaca
     3481 tcaagatgcc cggtatccat gccgttgatc accgcctgga aaggttagag gaaagtgaca
     3541 atgaaatgtt cgtagtacaa cgcgaacacg cagttgccaa gttcgccggg cttggtggtg
     3601 ggatggacga gctgtacaag tagtaactcg agatatctag acccagcttt cttgtacaaa
     3661 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     3721 caaaataaaa tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg
     3781 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     3841 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     3901 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     3961 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     4021 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     4081 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     4141 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     4201 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     4261 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     4321 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     4381 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     4441 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     4501 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     4561 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     4621 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     4681 ggttaattgg ttgtaacatt attcagattg ggccccgttc cactgagcgt cagaccccgt
     4741 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4801 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4861 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4921 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4981 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5041 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5101 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5161 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5221 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5281 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5341 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5401 tgctcacatg tt
//
