LOCUS       sequence-223548-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bimolecular Fluorescence Complimentation (BiFC) assay. Gateway 
            compatible N-terminal vector with BiFC-tag (HA-GFP1-10) to screen 
            interaction partners..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  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 7795)
  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: HA-GFP1-10-pDEST-N
FEATURES             Location/Qualifiers
     source          1..7795
                     /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"
     CDS             570..596
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1260..1384
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1409..1414,1415..1432,1433..1439)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1493..2152
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1559..1578)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2494..2799
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2678..2697
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2840..2964)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2996..3015)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2997..3015)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3172..3306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3209..3228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3263..3282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3432..3887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3519..3538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3729..3750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4006..4363
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4006..4197
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4029..4049)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4214..4349
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4414..5208
                     /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(4468..4487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5078..5097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5272..5320
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5361..5380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5480..5502
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5540..5558)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5626..5730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5731..5799,5800..6591)
                     /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(5949..5968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6762..7350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7251..7270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7504..7521
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 gctaccggtc gccaccatgt acccatacga tgttccagat tacgctgttt
      601 cgaaaggcga ggagctgttc acaggcgtgg tgccaatcct ggtggagctg gacggcgacg
      661 tgaacggcca caaattcagc gtgagaggcg agggcgaggg cgacgccaca atcggcaaac
      721 tgacactgaa attcatctgc acaacaggca aactgccagt gccctggcca acactagtga
      781 caacactgac atacggcgtg cagtgcttca gcagatatcc ggaccacatg aaaagacacg
      841 acttcttcaa aagcgccatg ccagagggct acgtgcagga gagaacaatc agcttcaaag
      901 acgacggcaa atacaaaaca agagccgtgg tgaaattcga gggcgacaca ctggtgaaca
      961 gaatcgagct gaaaggcaca gacttcaaag aggacggcaa catcctgggc cacaaactgg
     1021 agtacaactt caacagccac aacgtgtaca tcacagccaa caaacagaaa aacggcatca
     1081 aagccaactt cacagtgaga cacaacgtgg aggacggcag cgtgcagctc gccgaccact
     1141 accagcagaa cacaccaatc ggcgacggcc cagtgctgct gccagacaac cactacctga
     1201 gcacacagac agtgctgagc aaagacccaa acgagaaaag atctcgagct tctagatcaa
     1261 caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa
     1321 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac
     1381 tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt
     1441 gtggattttg agttaggatc cggcgagatt ttcaggagct aaggaagcta aaatggagaa
     1501 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     1561 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1621 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1681 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt
     1741 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     1801 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     1861 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     1921 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1981 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     2041 gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc atgtcggcag
     2101 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaagatctgg
     2161 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat
     2221 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtgtgc tatgaagcag
     2281 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca
     2341 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac
     2401 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg
     2461 gctcttttgc tgacgagaac agggactggt gaaatgcagt ttaaggttta cacctataaa
     2521 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg
     2581 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     2641 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     2701 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     2761 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac
     2821 agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt attatgtagt
     2881 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg
     2941 ttcagctttc ttgtacaaag tggtcgatgg ggtgcatcgg acgtcatagc tctctcccta
     3001 tagtgagtcg tattataagc taggcactgg ccgtcgttga tgggatccac cggatctaga
     3061 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3121 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3181 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3241 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3301 tggatcgatc ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3361 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3421 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3481 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3541 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3601 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3661 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3721 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3781 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3841 acaaatattt aacgcgaatt ttaacaaaat attaacgttt acaatttcgc ctgatgcggt
     3901 attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc
     3961 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     4021 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4081 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4141 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4201 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4261 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4321 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga
     4381 cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg attgcacgca
     4441 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4501 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4561 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4621 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4681 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4741 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4801 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4861 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4921 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4981 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5041 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5101 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5161 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5221 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata
     5281 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc
     5341 cgcgtatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5401 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5461 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5521 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5581 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5641 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5701 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5761 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5821 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5881 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5941 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6001 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6061 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6121 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6181 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6241 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6301 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6361 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6421 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6481 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6541 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6601 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6901 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7141 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7381 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7441 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7501 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7561 gcgcgttggc cgattcatta atgcagagct tgcaattcgc gcgtttttca atattattga
     7621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7681 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7741 attattatca tgacattaac ctataaaaat aggcgtagta cgaggccctt tcact
//
