LOCUS       sequence-445920-        8028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with Dok1 tagged with Clover.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8028)
  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 8028)
  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-Dok1-Clover
FEATURES             Location/Qualifiers
     source          1..8028
                     /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(3244..3246,3247..3249,3250..3960)
                     /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(3289..3310)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3897..3918
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3976..4000)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4026..4067
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4077..4094
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4117..4134)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4123..4347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4393..4821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4480..4499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4690..4711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4830..4850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4835..5164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5015..5150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5077..5096
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5212..5259
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5278..5676
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5834..5955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5871..5890)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5925..5944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6004..6020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6004..6020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6017..6039)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6028..6044
                     /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(6052..6058,6059..6076,6077..6082))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6097..6118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6235..6252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6395..6983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6475..6494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7154..7945,7946..8014))
                     /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     7777..7796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8015..8028,1..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 gcaccatgga cggagcagtg
     1801 atggaagggc cgcttttttt gcagagtcag cgctttggga ccaagaggtg gaggaagacc
     1861 tgggccgtgc tctacccggc cagtccccac ggcgtagcgc ggctcgagtt ctttgaccat
     1921 aaggggtcga gctctggggg tggccgaggg agctcgcgcc gcctggactg caaagtgatc
     1981 cgtctggctg agtgtgtgag tgtggccccc gtcaccgtgg agaccccccc tgagcccggc
     2041 gccactgcct tccgcctgga cactgctcag cgctcgcacc tgctggcggc cgacgcgccg
     2101 tccagtgcag cctgggtgca gacgctgtgc cgaaacgcct ttccgaaagg cagctggact
     2161 ctggcgccta ccgataaccc acctaagctt tctgccctgg agatgctgga gaactccttg
     2221 tacagcccta cctgggaagg atcccaattc tgggtaacgg tgcagaggac tgaggccgcc
     2281 gagcgctgtg gcctgcatgg ctcctacgtg ctgagggtgg aggctgaaag gctgactctc
     2341 ctgaccgtgg gggcccagag tcagatactg gagccactcc tgtcctggcc ctacactctg
     2401 ttgcgtcgct atggccggga caaggtcatg ttctctttcg aggccggccg ccgctgcccc
     2461 tcaggccctg gaaccttcac cttccagacg gcacagggaa atgacatctt ccaggcagtt
     2521 gagactgcca tccaccggca gaaggcccag ggaaaggccg gacaggggca cgatgttctc
     2581 agagctgact cccatgaagg ggaggtggca gaggggaagt tgccttcccc acctggcccc
     2641 caagagctcc tcgacagtcc cccagccctg tatgctgagc ccttagactc cctgcgcatt
     2701 gctccatgcc cttcccagga ctccctatac tcagacccct tggacagcac gtctgctcag
     2761 gcaggagagg gagtacaacg gaagaaacct ctctattggg acttgtatga gcatgcgcag
     2821 cagcagttgc tgaaggccaa gctgacagac cccaaagagg atcccatcta tgatgaacct
     2881 gagggcctgg ccccagtccc tccccagggc ctttatgatc tgcctcggga gcccaaggat
     2941 gcatggtggt gccaagctcg ggtgaaggag gagggctatg agctccccta caaccctgcc
     3001 actgatgact acgctgtgcc accccctcgg agcacaaagc ccctccttgc tcccaagccc
     3061 cagggcccag ccttccctga acctggtact gcaactggca gtggcatcaa aagccacaac
     3121 tcagccctgt acagccaggt ccagaagagc ggggcctcag ggagctggga ctgtgggctc
     3181 tctagagtag ggactgacaa gactggggtc aagtcagagg gctctaccgg cggcggcggc
     3241 cgcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3301 gacggcgacg taaacggcca caagttcagc gtccgcggcg agggcgaggg cgatgccacc
     3361 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3421 accctcgtga ccaccttcgg ctacggcgtg gcctgcttca gccgctaccc cgaccacatg
     3481 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3541 tctttcaagg acgacggtac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3601 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3661 cacaagctgg agtacaactt caacagccac aacgtctata tcacggccga caagcagaag
     3721 aacggcatca aggctaactt caagatccgc cacaacgttg aggacggcag cgtgcagctc
     3781 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3841 cactacctga gccatcagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3901 gtcctgctgg agttcgtgac cgccgccggg attacacatg gcatggacga gctgtacaag
     3961 tagtaagata tctagaccca gctttcttgt acaaagtggt tgatctagag ggcccgcggt
     4021 tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc
     4081 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     4141 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4201 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4261 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4321 atgctgggga tgcggtgggc gctatggctt ctgaggcgga aagaaccagc tggggctcta
     4381 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4441 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4501 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4561 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4621 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4681 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4741 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4801 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4861 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4981 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5041 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5101 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5161 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     5221 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     5281 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     5341 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     5401 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     5461 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     5521 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     5581 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     5641 gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg aggagcagga
     5701 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5761 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5821 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5881 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5941 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6001 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6061 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6121 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6181 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6241 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6301 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttc
//
