LOCUS       sequence-369579-        8163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mVenus tagged Focal Adhesion Kinase with FRB inserted at Y397.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8163)
  AUTHORS   Fauser J, Huyot V, Matsche J, Szynal BN, Alexeev Y, Kota P, Karginov
            AV
  TITLE     Dissecting protein tyrosine phosphatase signaling by engineered 
            chemogenetic control of its activity.
  JOURNAL   J Cell Biol. 2022 Aug 1;221(8). pii: 213352. doi: 
            10.1083/jcb.202111066. Epub 2022 Jul 13.
  PUBMED    35829702
REFERENCE   2  (bases 1 to 8163)
  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: "J Cell 
            Biol."; date: "2022-08-1"; volume: "221(8). pii: 213352. doi"; 
            pages: " 10.1083/jcb.202111066. Epub 2022 Jul 13"
COMMENT     Sequence Label: mVenus-FAK(397)FRB
FEATURES             Location/Qualifiers
     source          1..8163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4951..5072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4988..5007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5042..5061
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5079..5534)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5216..5237)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5428..5447
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5561..5665
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5667..6024
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5667..5858
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5690..5710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5875..6010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6059..6853
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6059..6853
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6113..6132)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6723..6742
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7041..7060)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7085..7132
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7461..8049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7950..7969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttgatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctcggc tacggcctgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggcaaacttc aagatccgcc acaacatcga ggacggcggc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctggaatg gcagctgctt atcttgaccc aaacttgaat
     1381 cacacaccaa gttcgagtac taagactcac ctgggtactg gcatggaacg gtcccctggt
     1441 gcaatggaac gagtattaaa ggtctttcat cattttgaaa gcagtagtga gccaaccacc
     1501 tgggccagta ttatcaggca tggagatgct actgatgtca ggggcatcat tcagaagata
     1561 gtggacagtc acaaagtaaa gcacgtggcc tgctatggat ttcgcctcag tcacctgcga
     1621 tcggaggagg tgcactggct gcacgtggac atgggtgtct ccagtgtgag ggagaaatat
     1681 gaactcgccc acccaccaga ggagtggaaa tatgagttga gaattcgtta cttgccaaaa
     1741 ggatttctaa accagtttac tgaagataag ccaacattga atttcttcta tcaacaggtg
     1801 aagagtgact acatgcaaga aatagctgat caagtagacc aagaaatagc tttgaagttg
     1861 ggttgtttgg aaattaggcg atcctattgg gagatgaggg gtaatgccct agagaagaag
     1921 tccaactatg aagtattaga aaaagatgtt ggtttaaagc gattttttcc taagagttta
     1981 ctggattctg tcaaggccaa aacactaaga aagctgatcc agcaaacatt caggcagttt
     2041 gccaacctta atagagaaga aagcatcctg aaattctttg agattctttc tcctgtgtac
     2101 agatttgaca aagagtgctt caagtgtgcc cttgggtcaa gttggatcat ttctgtggaa
     2161 ttggcaatcg gcccagaaga agggatcagt tacctgacag acaaaggctg caatcccaca
     2221 cacctagcag actttaacca agtgcagacc atccagtact caaacagtga agacaaagac
     2281 aggaaaggaa tgctacaact caaaatcgct ggtgctcctg agcctctgac cgtgacggca
     2341 ccatccctaa ccattgccga gaacatggcc gacctgattg atggatactg ccggctggtg
     2401 aatggagcca cacaatcttt catcatcaga ccccagaaag aaggtgaacg ggcgttgcca
     2461 tcaataccaa agttggccaa cagtgaaaag caaggcatgc ggacacatgc agtctctgtg
     2521 tcagagacag atgacggccc cggatggcat gagatgtggc atgaaggcct ggaagaggca
     2581 tctcgtttgt actttgggga aaggaacgtg aaaggcatgt ttgaggtgct ggagcccttg
     2641 catgctatga tggaacgggg cccccagact ctgaaggaaa catcctttaa tcaggcctat
     2701 ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt acatgaaatc agggaatgtc
     2761 aaggacctcc tccaagcctg ggacctctat tatcatgtgt tccgacgaat ctcaaagact
     2821 agtggacccg gtgcagagat catcgatgag gaagacacat acaccatgcc ctcgaccagg
     2881 gattatgaga ttcagagaga aagaatagaa cttggacgct gtattggaga aggtcagttt
     2941 ggagatgtac atcaaggcgt gtacctgagc ccagagaatc cagctttggc tgttgcaatc
     3001 aaaacatgta aaaactgtac ttcggacagc gtgagagaga agttccttca agaagcctta
     3061 acaatgcgtc agtttgacca tcctcacatc gtgaagctga ttggagtcat tacagagaac
     3121 cctgtctgga taatcatgga gttgtgcaca cttggagagc tgaggtcatt tttgcaagta
     3181 agaaaataca gcttggacct ggcatctttg atattatatg cctatcagct tagtacagca
     3241 ctcgcgtatc tggagagcaa aagatttgtt cacagggaca ttgctgctcg gaatgttctg
     3301 gtgtcttcaa atgattgtgt aaaattggga gactttggat tatctcgata tatggaagac
     3361 agtacttact ataaagcttc caaaggaaaa ttacctatta aatggatggc tccagagtca
     3421 atcaattttc gacgttttac ctcagctagt gacgtgtgga tgtttggtgt gtgtatgtgg
     3481 gagatactga tgcatggcgt gaagcctttt caaggagtga agaacaatga tgtgatcggt
     3541 cgaattgaaa atggggaaag attaccaatg cctccaaatt gtcctcccac cctctacagc
     3601 cttatgacga aatgttgggc ctatgacccc agcaggcggc ccaggtttac tgaactaaaa
     3661 gctcagctca gcacaatcct ggaggaggag aaggtgcagc aagaagaacg gatgaggatg
     3721 gaatccagaa gacaggctac cgtgtcctgg gactctggag ggtctgatga agcaccaccc
     3781 aagcccagca gacctggtta tcctagcccg agatccagtg aaggatttta tcccagccca
     3841 cagcacatgg tccagaccaa tcactaccag gtctctggct acccgggttc ccatggaatc
     3901 ccagccatgg ctggcagcat ttaccaaggt caggcatctc ttctggacca gacagaattg
     3961 tggaatcaca gacctcagga gatgtccatg tggcagccca gtgtggagga ttcggcagct
     4021 ttggaccttc gagggatggg acaggtgctt ccccctcacc tgatggaaga gcggctaatc
     4081 cgacagcagc aggaaatgga agaagaccag cgctggctgg agaaggaaga gcgattcctg
     4141 aaacctgatg tgaggctctc tcgaggcagc atcgacaggg aagacgggag ttttcagggt
     4201 ccgactggaa accaacacat ctatcagcct gtggggaagc cagatcctgc agctccacca
     4261 aagaaacctc ctcgccctgg agcacctggc cacctaagca acctgtccag catcagcagc
     4321 cctgcagaca gctacaatga gggtgtcaag cttcagcccc aggaaatcag cccccctccc
     4381 actgccaacc ttgaccggtc caatgacaag gtatatgaaa atgtgacagg cctagtgaag
     4441 gctgtcatcg agatgtccag caaaatccag ccagctcctc cagaagagta cgtccctatg
     4501 gtgaaggaag ttggcctggc cctgcggacc ttactagcca cggtggatga gaccattcct
     4561 gctcttccag ccagcactca tcgagagatc gagatggcac agaagctgct gaactccgac
     4621 ttaggcgagc tcatcagcaa gatgaagctg gcgcagcagt acgtcatgac cagcctgcag
     4681 caggagtata agaagcagat gctgacagct gctcacgccc tggctgtgga tgccaagaac
     4741 ctacttgatg ttattgatca agcaagactg aaaatgctag ggcagacacg gccacactga
     4801 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     4861 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4921 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4981 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5041 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     5101 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5161 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5221 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5281 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5341 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5401 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5461 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5521 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5581 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5641 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5701 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5761 ctcaattagt cagcaaccag gtgtggaaag tccccatgct ccccagcagg cagaagtatg
     5821 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5881 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5941 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6001 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     6061 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     6121 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     6181 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     6241 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     6301 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     6361 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     6421 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     6481 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     6541 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     6601 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6661 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6721 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6781 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6841 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6901 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6961 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7021 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     7081 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     7141 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     7201 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     7261 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     7321 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7381 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7441 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7501 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7561 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7621 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7681 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7741 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7801 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7861 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7921 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7981 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8041 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8101 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     8161 cat
//
