LOCUS       sequence-485186-        6223 bp    DNA     circular SYN 03-SEP-2025
DEFINITION  Expression vector with integrin beta1 tagged with the BiFC V1 
            fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  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 6223)
  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: pDEST-ITGB1-V1
FEATURES             Location/Qualifiers
     source          1..6223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    942..966
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3406..3430)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3474..3476,3477..3935)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(3516..3537)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3777..3796)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3979..3996)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3985..4209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4297..4314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4468..5056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4548..4567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5227..6018,6019..6087))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5850..5869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6088..6192)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccatattacg gggtactcga gacaagtttg tacaaaaaag
      961 caggctggcg ccgaccaatt cagtcgactg gatccggtac caccgccatg aatttacaac
     1021 caattttctg gattggactg atcagttcag tttgctgtgt gtttgctcaa acagatgaaa
     1081 atagatgttt aaaagcaaat gccaaatcat gtggagaatg tatacaagca gggccaaatt
     1141 gtgggtggtg cacaaattca acatttttac aggaaggaat gcctacttct gcacgatgtg
     1201 atgatttaga agccttaaaa aagaagggtt gccctccaga tgacatagaa aatcccagag
     1261 gctccaaaga tataaagaaa aataaaaatg taaccaaccg tagcaaagga acagcagaga
     1321 agctcaagcc agaggatatt actcagatcc aaccacagca gttggttttg cgattaagat
     1381 caggggagcc acagacattt acattaaaat tcaagagagc tgaagactat cccattgacc
     1441 tctactacct tatggacctg tcttactcaa tgaaagacga tttggagaat gtaaaaagtc
     1501 ttggaacaga tctgatgaat gaaatgagga ggattacttc ggacttcaga attggatttg
     1561 gctcatttgt ggaaaagact gtgatgcctt acattagcac aacaccagct aagctcagga
     1621 acccttgcac aagtgaacag aactgcacca gcccatttag ctacaaaaat gtgctcagtc
     1681 ttactaataa aggagaagta tttaatgaac ttgttggaaa acagcgcata tctggaaatt
     1741 tggattctcc agaaggtggt ttcgatgcca tcatgcaagt tgcagtttgt ggatcactga
     1801 ttggctggag gaatgttaca cggctgctgg tgttttccac agatgccggg tttcactttg
     1861 ctggagatgg gaaacttggt ggcattgttt taccaaatga tggacaatgt cacctggaaa
     1921 ataatatgta cacaatgagc cattattatg attatccttc tattgctcac cttgtccaga
     1981 aactgagtga aaataatatt cagacaattt ttgcagttac tgaagaattt cagcctgttt
     2041 acaaggagct gaaaaacttg atccctaagt cagcagtagg aacattatct gcaaattcta
     2101 gcaatgtaat tcagttgatc attgatgcat acaattccct ttcctcagaa gtcattttgg
     2161 aaaacggcaa attgtcagaa ggagtaacaa taagttacaa atcttactgc aagaacgggg
     2221 tgaatggaac aggggaaaat ggaagaaaat gttccaatat ttccattgga gatgaggttc
     2281 aatttgaaat tagcataact tcaaataagt gtccaaaaaa ggattctgac agctttaaaa
     2341 ttaggcctct gggctttacg gaggaagtag aggttattct tcagtacatc tgtgaatgtg
     2401 aatgccaaag cgaaggcatc cctgaaagtc ccaagtgtca tgaaggaaat gggacatttg
     2461 agtgtggcgc gtgcaggtgc aatgaagggc gtgttggtag acattgtgaa tgcagcacag
     2521 atgaagttaa cagtgaagac atggatgctt actgcaggaa agaaaacagt tcagaaatct
     2581 gcagtaacaa tggagagtgc gtctgcggac agtgtgtttg taggaagagg gataatacaa
     2641 atgaaattta ttctggcaaa ttctgcgagt gtgataattt caactgtgat agatccaatg
     2701 gcttaatttg tggaggaaat ggtgtttgca agtgtcgtgt gtgtgagtgc aaccccaact
     2761 acactggcag tgcatgtgac tgttctttgg atactagtac ttgtgaagcc agcaacgcac
     2821 agatctgcaa tggccggggc atctgcgagt gtggtgtctg taagtgtaca gatccgaagt
     2881 ttcaagggca aacgtgtgag atgtgtcaga cctgccttgg tgtctgtgct gagcataaag
     2941 aatgtgttca gtgcagagcc ttcaataaag gagaaaagaa agacacatgc acacaggaat
     3001 gttcctattt taacattacc aaggtagaaa gtcgggacaa attaccccag ccggtccaac
     3061 ctgatcctgt gtcccattgt aaggagaagg atgttgacga ctgttggttc tattttacgt
     3121 attcagtgaa tgggaacaac gaggtcatgg ttcatgttgt ggagaatcca gagtgtccca
     3181 ctggtccaga catcattcca attgtagctg gtgtggttgc tggaattgtt cttattggcc
     3241 ttgcgttact cctcatttgg aagcttttaa tgataattca tgacagaagg gagtttgcta
     3301 aatttgaaaa ggagaaaatg aatgccaaat gggacacggg tgaaaatcct atttataaga
     3361 gtgccgtaac aactgtggtc aatccgaagt atgagggaaa acgctaccca gctttcttgt
     3421 acaaagtggt catcgatggt ggcggtggct ctggaggtgg tgggtcctcc ggagtgagca
     3481 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3541 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     3601 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3661 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     3721 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3781 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3841 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3901 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagtaatct agagggcccg
     3961 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4021 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4081 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4141 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4201 gctctatggc ttctgaggcg gaaagaacca gctgcattaa tgaatcggcc aacgcgcggg
     4261 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5641 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5881 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6061 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6121 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6181 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
