LOCUS       sequence-233678-        7771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bimolecular Fluorescence Complimentation (BiFC) assay. Gateway 
            compatible C-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 7771)
  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 7771)
  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-C
FEATURES             Location/Qualifiers
     source          1..7771
                     /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"
     CDS             2357..2383
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3151..3285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3188..3207)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3242..3261
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3411..3866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3498..3517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3708..3729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3983..4340
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3983..4174
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4006..4026)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4191..4326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4391..5185
                     /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(4445..4464)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5055..5074
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5249..5297
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5338..5357)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5457..5479
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5603..5707
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5708..5776,5777..6568)
                     /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(5926..5945)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6739..7327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7228..7247
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7481..7498
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(7572..7676)
                     /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 tccatgtacc catacgatgt tccagattac gctgtttcga aaggcgagga
     2401 gctgttcaca ggcgtggtgc caatcctggt ggagctggac ggcgacgtga acggccacaa
     2461 attcagcgtg agaggcgagg gcgagggcga cgccacaatc ggcaaactga cactgaaatt
     2521 catctgcaca acaggcaaac tgccagtgcc ctggccaaca ctagtgacaa cactgacata
     2581 cggcgtgcag tgcttcagca gatatccgga ccacatgaaa agacacgact tcttcaaaag
     2641 cgccatgcca gagggctacg tgcaggagag aacaatcagc ttcaaagacg acggcaaata
     2701 caaaacaaga gccgtggtga aattcgaggg cgacacactg gtgaacagaa tcgagctgaa
     2761 aggcacagac ttcaaagagg acggcaacat cctgggccac aaactggagt acaacttcaa
     2821 cagccacaac gtgtacatca cagccaacaa acagaaaaac ggcatcaaag ccaacttcac
     2881 agtgagacac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2941 accaatcggc gacggcccag tgctgctgcc agacaaccac tacctgagca cacagacagt
     3001 gctgagcaaa gacccaaacg agaaataaag cggccgcgac tctagatcat aatcagccat
     3061 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3121 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3181 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3241 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgatcc tgcattaatg
     3301 aatcggccaa cgcgcgggga gaggcggttt gcgtattggc tggcgtaata gcgaagaggc
     3361 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     3421 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3481 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3541 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3601 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3661 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3721 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3781 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3841 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     3901 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     3961 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4021 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4201 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4261 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4321 gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaaggc
     4381 tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4441 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4501 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4561 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4621 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4681 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4741 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4801 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4861 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4921 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4981 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5041 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5101 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5161 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5221 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5281 tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca ctctcagtac
     5341 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5401 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5461 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5521 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5581 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5641 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5701 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5761 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5821 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5881 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5941 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6001 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6061 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6121 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6181 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6241 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6301 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6361 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6421 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6481 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6541 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6601 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6661 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6721 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6781 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6841 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6901 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6961 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7021 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7081 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7141 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7201 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7261 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7321 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7381 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     7441 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7501 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     7561 cagagcttgc aattcgcgct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7681 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7741 aaaaataggc gtattacgag gccctttcac g
//
