LOCUS       sequence-446286-        8368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with clover-tagged Shp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8368)
  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 8368)
  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: pEF.DEST51-PTPN11-Clover
FEATURES             Location/Qualifiers
     source          1..8368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(166..185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        473..1651
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          704..1642
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1599..1619
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1668..1687
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1668..1686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1718..1742
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3584..3586,3587..3589,3590..4300)
                     /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(3629..3650)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4237..4258
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4316..4340)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4366..4407
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4417..4434
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4457..4474)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4463..4687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4733..5161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4820..4839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5030..5051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5170..5190)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5175..5504
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5355..5490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5417..5436
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5552..5599
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5618..6016
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6174..6295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6211..6230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6265..6284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6344..6360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6344..6360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6357..6379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6368..6384
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6392..6398,6399..6416,6417..6422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6437..6458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6575..6592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6735..7323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6815..6834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7494..8285,8286..8354))
                     /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     8117..8136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8355..8368,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
      481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
      721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
      781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
      841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
      901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
      961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     1681 tatagggaga cccaagctgg ctaggtaagc ttgatcaaca agtttgtaca aaaaagcagg
     1741 ctggcgccgg aaccaattca gtcgactgga tccggtaccg aggagatctg ccgccgcgat
     1801 cgccatgaca tcgcggagat ggtttcaccc aaatatcact ggtgtggagg cagaaaacct
     1861 actgttgaca agaggagttg atggcagttt tttggcaagg cctagtaaaa gtaaccctgg
     1921 agacttcaca ctttccgtta gaagaaatgg agctgtcacc cacatcaaga ttcagaacac
     1981 tggtgattac tatgacctgt atggagggga gaaatttgcc actttggctg agttggtcca
     2041 gtattacatg gaacatcacg ggcaattaaa agagaagaat ggagatgtca ttgagcttaa
     2101 atatcctctg aactgtgcag atcctacctc tgaaaggtgg tttcatggac atctctctgg
     2161 gaaagaagca gagaaattat taactgaaaa aggaaaacat ggtagttttc ttgtacgaga
     2221 gagccagagc caccctggag attttgttct ttctgtgcgc actggtgatg acaaagggga
     2281 gagcaatgac ggcaagtcta aagtgaccca tgttatgatt cgctgtcagg aactgaaata
     2341 cgacgttggt ggaggagaac ggtttgattc tttgacagat cttgtggaac attataagaa
     2401 gaatcctatg gtggaaacat tgggtacagt actacaactc aagcagcccc ttaacacgac
     2461 tcgtataaat gctgctgaaa tagaaagcag agttcgagaa ctaagcaaat tagctgagac
     2521 cacagataaa gtcaaacaag gcttttggga agaatttgag acactacaac aacaggagtg
     2581 caaacttctc tacagccgaa aagagggtca aaggcaagaa aacaaaaaca aaaatagata
     2641 taaaaacatc ctgccctttg atcataccag ggttgtccta cacgatggtg atcccaatga
     2701 gcctgtttca gattacatca atgcaaatat catcatgcct gaatttgaaa ccaagtgcaa
     2761 caattcaaag cccaaaaaga gttacattgc cacacaaggc tgcctgcaaa acacggtgaa
     2821 tgacttttgg cggatggtgt tccaagaaaa ctcccgagtg attgtcatga caacgaaaga
     2881 agtggagaga ggaaagagta aatgtgtcaa atactggcct gatgagtatg ctctaaaaga
     2941 atatggcgtc atgcgtgtta ggaacgtcaa agaaagcgcc gctcatgact atacgctaag
     3001 agaacttaaa ctttcaaagg ttggacaagg gaatacggag agaacggtct ggcaatacca
     3061 ctttcggacc tggccggacc acggcgtgcc cagcgaccct gggggcgtgc tggacttcct
     3121 ggaggaggtg caccataagc aggagagcat catggatgca gggccggtcg tggtgcactg
     3181 cagtgctgga attggccgga cagggacgtt cattgtgatt gatattctta ttgacatcat
     3241 cagagagaaa ggtgttgact gcgatattga cgttcccaaa accatccaga tggtgcggtc
     3301 tcagaggtca gggatggtcc agacagaagc acagtaccga tttatctata tggcggtcca
     3361 gcattatatt gaaacactac agcgcaggat tgaagaagag cagaaaagca agaggaaagg
     3421 gcacgaatat acaaatatta agtattctct agcggaccag acgagtggag atcagagccc
     3481 tctcccgcct tgtactccaa cgccaccctg tgcagaaatg agagaagaca gtgctagagt
     3541 ctatgaaaac gtgggcctga tgcaacagca gaaaagcggc cgcatggtga gcaagggcga
     3601 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3661 caagttcagc gtccgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     3721 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg
     3781 ctacggcgtg gcctgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3841 gtccgccatg cccgaaggct acgtccagga gcgcaccatc tctttcaagg acgacggtac
     3901 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3961 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     4021 caacagccac aacgtctata tcacggccga caagcagaag aacggcatca aggctaactt
     4081 caagatccgc cacaacgttg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4141 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gccatcagtc
     4201 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     4261 cgccgccggg attacacatg gcatggacga gctgtacaag tagtaagata tctagaccca
     4321 gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa gcctatccct
     4381 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt
     4441 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4501 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4561 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4621 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4681 gctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4741 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4801 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4861 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4921 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4981 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5041 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5101 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5161 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5221 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5281 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5341 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5401 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5461 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5521 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     5581 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga
     5641 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     5701 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     5761 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     5821 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     5881 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     5941 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     6001 tgtgtgggag ggctaagcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     6061 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6121 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6181 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6241 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6301 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6361 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6421 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6481 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6541 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6601 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6661 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccaggaaccg taaaaaggcc
     6721 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6781 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6841 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6901 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6961 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7021 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7081 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7141 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7501 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7561 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7621 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7681 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7741 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7801 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7861 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7921 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7981 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8041 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8101 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8161 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8221 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8281 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8341 tgttgaatac tcatactctt cctttttc
//
