LOCUS       sequence-496365-        6175 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Mammalian expression vector of YAP fused with mScarlet3-H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6175)
  AUTHORS   Nishimura R, Barnett SFH, Jain K, Huang Z, Goult BT, Kanchanawong P
  TITLE     Optogenetic control of mechanotransduction based on light-induced 
            homodimerization of talin.
  JOURNAL   J Cell Sci. 2025 Nov 24:jcs.264110. doi: 10.1242/jcs.264110.
  PUBMED    41277405
REFERENCE   2  (bases 1 to 6175)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell Sci.
            2025 Nov 24:jcs.264110. doi: 10.1242/jcs.264110."
COMMENT     Sequence Label: pEGFP-N1/YAP-mScarlet3-H
FEATURES             Location/Qualifiers
     source          1..6175
                     /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      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2151..2837
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     2549..2568
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2963..3084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3000..3019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3054..3073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3091..3546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3228..3249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3440..3459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3573..3677
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3679..4036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3679..3870
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3702..3722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3887..4022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4071..4865
                     /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             4071..4865
                     /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(4125..4144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4735..4754
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5053..5072)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5097..5144
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5473..6061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5962..5981
                     /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 gctagcgcca
      601 ccatggagcc cgcgcaacag ccgccgcccc agccggcccc gcaaggcccc gcgccgccgt
      661 ccgtgtctcc ggccgggacc cccgcggccc cgcccgcacc cccggccggc caccaggtcg
      721 tgcacgtccg cggggactcg gagaccgact tggaggcgct cttcaatgcc gtcatgaacc
      781 ccaagacggc caacgtgcct cagaccgtgc ccatgcggct tcgcaagctg cccgactcct
      841 tcttcaagcc gcctgagccc aagtcccact cgcgacaggc cagtactgat gcaggtactg
      901 cgggagctct gactccacag catgttcgag ctcactcctc tccagcctcc ctgcagctgg
      961 gtgccgtttc tcctgggaca ctcacagcca gtggcgttgt ctctggccct gccgctgccc
     1021 ctgcagctca gcatctccgg cagtcctcct ttgagatccc tgatgatgta ccactgccag
     1081 caggctggga gatggccaag acatcttctg gtcaaagata cttcttaaat cacaacgatc
     1141 agacaacaac atggcaggac ccccggaagg ccatgctttc gcaactgaac gttcctgcgc
     1201 ctgccagccc agcggtgccc cagacgctga tgaattctgc ctcaggacct cttcctgatg
     1261 gatgggagca agccatgact caggatggag aagtttacta cataaaccat aagaacaaga
     1321 ccacatcctg gctggaccca aggctggacc ctcgttttgc catgaaccag aggatcactc
     1381 agagtgctcc agtgaagcag cccccaccct tggctcccca gagcccacag ggaggcgtcc
     1441 tgggtggagg cagttccaac cagcagcagc aaatacagct gcagcagtta cagatggaga
     1501 aggagagact gcggttgaaa caacaggaat tatttcggca ggcaatacgg aatatcaatc
     1561 ccagcacagc aaatgctcca aaatgtcagg aattagctct gcgcagccag ttgcctacac
     1621 tggagcagga tggagggact ccgaatgcag tgtcttctcc tgggatgtct caggaattga
     1681 gaacaatgac aaccaatagt tccgatccct ttcttaacag tggcacctat cactctcgag
     1741 atgagagcac agacagcggc ctcagcatga gcagctacag catccctcgg accccagacg
     1801 acttcctcaa cagtgtggat gaaatggata caggagacac catcagccaa agcaccctgc
     1861 cgtcacagca gagccgcttc cccgactacc tggaagccct ccctgggaca aatgtggacc
     1921 ttggcacact ggaaggagat gcaatgaaca tagaagggga ggagctgatg cccagtctgc
     1981 aggaagcgct gagttccgaa atcttggacg tggagtctgt gttggctgcc accaagctag
     2041 ataaagaaag ctttctcacg tggttaaagc ttcgaattct gcagtcgacg gtaccgcggg
     2101 atccatctgg tggcggaggg agtggcggag gtgggtcagg aggtggaggc atggatagca
     2161 ccgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc tccatgaacg
     2221 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2281 ccaagctgag ggtgaccaag ggtggccccc tgcccttctc ctgggacatc ctgtcccctc
     2341 agttcatgta cggctccagg gccttcacga agcaccccgc cgacatcccc gactactgga
     2401 agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     2461 ccgtgtccgt ggcccaggac acctccctgg aggacggcac cctgatctac aaggtgaagc
     2521 tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca atgggctggg
     2581 aagcatccac cgagcggttg taccccgagg acgtcgtgct gaagggcgac attaagcatg
     2641 ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc tacagggcca
     2701 agaagcccgt gcagatgccc ggcgccttca acatcgaccg caagttggac atcacatccc
     2761 acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgtggcccgc cactccaccg
     2821 gcggctccgg tggctcctaa agcggccgcg actctagatc ataatcagcc ataccacatt
     2881 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2941 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3061 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3121 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3181 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3241 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3301 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3361 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3421 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3481 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3541 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3601 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3661 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3781 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3841 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3901 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3961 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4021 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4081 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4141 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4201 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4261 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4321 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4381 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4441 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4501 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4561 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4621 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4681 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4741 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4801 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4861 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4921 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4981 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5041 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5101 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5161 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5221 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5281 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5341 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5401 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5461 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5521 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5581 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5641 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5701 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5761 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5821 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5881 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5941 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6001 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6061 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6121 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
