LOCUS       sequence-229338-        7227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bimolecular Fluorescence Complimentation (BiFC) assay. Gateway 
            compatible C-terminal vector with BiFC-tag (GFP11-Flag) to screen 
            interaction partners..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7227)
  AUTHORS   Viita T, Kyheroinen S, Prajapati B, Virtanen J, Frilander MJ, 
            Varjosalo M, Vartiainen MK
  TITLE     Nuclear actin interactome analysis links actin to KAT14 histone 
            acetyl transferase and mRNA splicing.
  JOURNAL   J Cell Sci. 2019 Apr 17;132(8). pii: jcs.226852. doi: 
            10.1242/jcs.226852.
  PUBMED    30890647
REFERENCE   2  (bases 1 to 7227)
  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 
            Sci."; date: "2019-04-17"; volume: "132(8). pii: jcs.226852. doi"; 
            pages: " 10.1242/jcs.226852"
COMMENT     Sequence Label: Flag-GFP11-pDEST-C
FEATURES             Location/Qualifiers
     source          1..7227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     473..493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     580..599
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        580..598
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    631..755
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(780..785,786..803,804..810)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             864..1544
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(930..949)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1864..2169
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2048..2067
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2210..2334)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2363..2372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2399..2446
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             2456..2479
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2607..2741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2644..2663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2698..2717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2867..3322
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2954..2973)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3164..3185
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3439..3796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3439..3630
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3462..3482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3647..3782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3847..4641
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3901..3920)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4511..4530
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4705..4753
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4794..4813)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4913..4935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5059..5163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5164..5232,5233..6024)
                     /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     complement(5382..5401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6195..6783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6684..6703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6937..6954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(7028..7132)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      541 atccgctagc gctaccggac tcagatctcg agctgaaatt aatacgactc actataggga
      601 gacccaagct ggctagttaa gcttgatcaa acaagtttgt acaaaaaagc tgaacgagaa
      661 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
      721 taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct
      781 ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat
      841 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
      901 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
      961 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     1021 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     1081 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     1141 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     1201 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     1261 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     1321 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     1381 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     1441 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     1501 atgagtggca ggcggggcgt aatctagagg atccggctta ctaaaagcca gataacagta
     1561 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
     1621 tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc
     1681 tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc
     1741 agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg
     1801 ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt
     1861 gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     1921 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
     1981 cgtctgctgt cagataaagt cccccgtgaa ctttacccgg tggtgcatat cggggatgaa
     2041 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     2101 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     2161 ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc atagtgactg
     2221 gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat
     2281 attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag tggttcgatc
     2341 ttacacggga tccaagccgg tcgccaccat gggcgacggc ggcagcggcg gcggcagccg
     2401 ggaccacatg gtgctgcacg agtacgtgaa cgccgccggc atcacatctg gagtggacta
     2461 caaggacgac gatgacaagg gataagcggc cgcgactcta gatcataatc agccatacca
     2521 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2581 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2641 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2701 gtttgtccaa actcatcaat gtatcttatc atgtctggat cgatcctgca ttaatgaatc
     2761 ggccaacgcg cggggagagg cggtttgcgt attggctggc gtaatagcga agaggcccgc
     2821 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     2881 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2941 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3001 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3061 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3121 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3181 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3241 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3301 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3361 tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg
     3421 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3541 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3601 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3661 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3721 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3781 cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taaggctaga
     3841 gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3901 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3961 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4021 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4081 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4141 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4201 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4261 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4321 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4381 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4441 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4501 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4561 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4621 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4681 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     4741 ggttttttgt gtgaatcgat agcgataagg atccgcgtat ggtgcactct cagtacaatc
     4801 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4861 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4921 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4981 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5041 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5101 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5401 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5701 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6961 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcaga
     7021 gcttgcaatt cgcgcttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7081 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7141 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     7201 ataggcgtat tacgaggccc tttcacg
//
