LOCUS       sequence-160931-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p63 (619 a.a.) tagged with mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   van Unen J, Yin T, Wu YI, Mastop M, Gadella TW Jr, Goedhart J
  TITLE     Kinetics of recruitment and allosteric activation of ARHGEF25 
            isoforms by the heterotrimeric G-protein Galphaq.
  JOURNAL   Sci Rep. 2016 Nov 11;6:36825. doi: 10.1038/srep36825.
  PUBMED    27833100
REFERENCE   2  (bases 1 to 6580)
  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: "Sci Rep."; 
            date: "2016-11-11"; volume: "6:36825. doi"; pages: " 
            10.1038/srep36825"
COMMENT     Sequence Label: p63RhoGEF619-mVenus-N1
FEATURES             Location/Qualifiers
     source          1..6580
                     /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      1332..1341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2520..2529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2526..2528,2529..2531,2532..3245)
                     /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(2571..2592)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2832..2851)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3179..3200
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3368..3489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3405..3424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3459..3478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3496..3951)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3633..3654)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3845..3864
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3978..4082
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4084..4441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4084..4275
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4107..4127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4292..4427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4476..5270
                     /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             4476..5270
                     /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(4530..4549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5140..5159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5458..5477)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5502..5549
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5878..6466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6367..6386
                     /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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc accatgaagc
      661 ccccggaccg ccccgcccct ggccgcactg accggatact gggggtcatg gggggcatgc
      721 tgcgcgcatg cgccctccct gggcaggagg ggcccccaag gagaagccct ctagggttgg
      781 tgggtaccga gccagagtct gaacgtacgg agggagatca cagaagggat cgcgaacatg
      841 aggtcctcgc cggggctctg cagcccgaat cctattccat tgcgggcagt gaggggagta
      901 tatcggcttc tgctgcctcc ggtctggctg ccccctctgg ccccagctct ggcctcagct
      961 ctggcccctg ttccccaggc cccccagggc ccgtcagtgg cctgaggaga tggttggatc
     1021 attccaaaca ttgtctcagt gtggaaactg aggcagacag tggtcaggca ggaccatatg
     1081 agaactggat gttggagcca gctctagcca caggagagga gctgccggaa ctgaccttgc
     1141 tgaccacact gttggagggc cctggagata agacgcagcc acctgaagag gagactttgt
     1201 cccaagcccc tgagagtgag gaggaacaga agaagaaggc tctggaaagg agtatgtatg
     1261 tcctgagtga actggtagaa acagagaaaa tgtacgtgga cgacttgggg cagattgtgg
     1321 agggttatat ggccaccatg gctgctcagg gggtccccga gagtcttcga ggccgtgaca
     1381 ggattgtgtt tgggaatatc cagcaaatct atgagtggca ccgagactat ttcttgcaag
     1441 agctacaacg gtgtctgaaa gatcctgatt ggctggctca gctattcatc aaacacgagc
     1501 gccggctgca tatgtatgtg gtgtactgtc agaataagcc caagtcagag catgtggtgt
     1561 cagagtttgg ggacagctac tttgaggagc tccggcagca gctggggcac cgcctgcagc
     1621 tgaacgacct cctcatcaaa cctgtgcagc ggatcatgaa ataccagctg ctgctcaagg
     1681 attttctcaa gtattacaat agagctggga tggatactgc agacctagag caagctgtgg
     1741 aggtcatgtg ctttgtgccc aagcgctgca acgatatgat gacgctgggg agattgcggg
     1801 gatttgaggg caaactgact gctcagggga agctcttggg ccaggacact ttctgggtca
     1861 ccgagcctga ggctggaggg ctgctgtctt cccgaggtcg agagaggcgc gtcttcctct
     1921 ttgagcaaat catcatcttc agtgaagccc tgggaggagg agtgagaggt ggaacacagc
     1981 ctggatatgt atacaagaac agcattaagg tgagctgcct gggactggag gggaacctcc
     2041 aaggtgaccc ttgccgcttt gcactgacct ccagagggcc agagggtggg atccagcgct
     2101 atgtcctgca ggctgcagac cctgctatca gtcaggcctg gatcaagcat gtggctcaga
     2161 tcttggagag ccaacgggac ttcctcaacg cattgcagtc acccattgag taccagagac
     2221 gggagagcca gaccaacagc ctggggcggc caagagggcc tggagtgggg agccctggaa
     2281 gaattcggct tggagatcag gcccagggca gcacacacac acccatcaat ggctctctcc
     2341 cctctctgct gctgtcaccc aaaggggagg tggccagagc cctcttgcca ctggataaac
     2401 aggcccttgg tgacatcccc caggctcccc atgactctcc tccagtctct ccaactccaa
     2461 aaacccctcc ctgccaagcc agacttgcca agctggatga agatgagctg tcaccggtcg
     2521 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2581 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2641 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     2701 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     2761 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2821 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2881 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2941 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     3001 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     3061 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3121 accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     3181 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3241 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3301 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3361 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3421 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3481 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3541 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3601 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3661 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3721 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3781 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3841 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3901 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3961 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4021 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4081 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4141 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4261 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4321 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4381 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4441 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4501 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4561 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4621 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4681 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4741 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4801 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4861 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4921 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4981 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5041 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5101 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5161 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5221 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5281 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5341 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5401 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5461 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5521 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5581 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5641 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5701 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5761 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5821 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5881 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5941 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6001 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     6061 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6121 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6181 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6241 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6301 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6361 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6421 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6481 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6541 atcccctgat tctgtggata accgtattac cgccatgcat
//
