LOCUS       sequence-471958-        7656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with Shp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7656)
  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 7656)
  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
FEATURES             Location/Qualifiers
     source          1..7656
                     /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"
     protein_bind    complement(3604..3628)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3654..3695
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3705..3722
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3745..3762)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3751..3975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4021..4449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4108..4127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4318..4339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4458..4478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4463..4792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4643..4778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4705..4724
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4840..4887
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4906..5304
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5462..5583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5499..5518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5553..5572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5632..5648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5632..5648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5645..5667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5656..5672
                     /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(5680..5686,5687..5704,5705..5710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5725..5746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5863..5880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6023..6611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6103..6122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6782..7573,7574..7642))
                     /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     7405..7424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7643..7656,1..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 cgctgataac tcgagatatc
     3601 tagacccagc tttcttgtac aaagtggttg atctagaggg cccgcggttc gaaggtaagc
     3661 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     3721 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3781 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3841 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3901 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3961 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     4021 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4081 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4141 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4201 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4261 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4321 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4381 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4441 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4501 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4561 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4621 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4681 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4741 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4801 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     4861 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagcctttg
     4921 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     4981 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     5041 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     5101 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     5161 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     5221 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5281 tctggttatg tgtgggaggg ctaagcactt cgtggccgag gagcaggact gacacgtgct
     5341 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5401 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5461 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5521 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5581 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5641 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5701 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5761 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5821 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5881 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5941 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc aggaaccgta
     6001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6841 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6901 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6961 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7021 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7081 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7141 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7201 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7261 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7321 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7381 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7441 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7501 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7561 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7621 cacggaaatg ttgaatactc atactcttcc tttttc
//
