LOCUS       sequence-229405-        7801 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 7801)
  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 7801)
  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..7801
                     /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    1269..1393
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1418..1423,1424..1441,1442..1448)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1502..2161
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1568..1587)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2503..2808
                     /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     2687..2706
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2849..2973)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3005..3024)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3006..3024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3181..3315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3272..3291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3441..3896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3528..3547)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3738..3759
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4013..4370
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4013..4204
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4036..4056)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4221..4356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4421..5215
                     /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(4475..4494)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5085..5104
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5279..5327
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5368..5387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5487..5509
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5633..5737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5738..5806,5807..6598)
                     /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(5956..5975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6769..7357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7258..7277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7511..7528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(7602..7706)
                     /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 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 acgagaaatc cggactcaga tctcgagctt
     1261 ctagatcaac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
     1321 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     1381 tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc
     1441 gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa
     1501 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1561 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1621 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1681 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     1741 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     1801 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     1861 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     1921 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     1981 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2041 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2101 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2161 aagatctgga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     2221 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct
     2281 atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat
     2341 atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc
     2401 gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg
     2461 aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac
     2521 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     2581 acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     2641 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     2701 gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc
     2761 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     2821 cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagta
     2881 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     2941 cgtttctcgt tcagctttct tgtacaaagt ggttgatcgc gtgcatgcga cgtcatagct
     3001 ctctccctat agtgagtcgt attataagct aggcactggc cgtcgttgat gggatccacc
     3061 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3121 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3181 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3241 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3301 tatcatgtct ggatcgatcc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3361 gcgtattggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3421 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3481 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3541 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3601 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3661 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3721 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3781 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3841 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     3901 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg
     3961 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     4021 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4141 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4201 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4261 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4321 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt
     4381 cttctgacac aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt
     4441 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4501 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4561 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4621 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4681 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4741 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4801 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4861 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4921 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4981 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5041 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5101 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5161 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5221 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa
     5281 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat
     5341 aaggatccgc gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5401 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5461 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5521 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5581 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5641 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5701 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5761 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5821 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5881 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5941 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6001 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6061 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6121 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6181 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6241 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6301 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6361 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6421 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6481 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6541 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6601 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6661 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6721 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6781 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6841 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6901 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6961 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7021 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7081 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7141 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7201 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7261 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7321 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7381 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7441 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7501 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7561 tctccccgcg cgttggccga ttcattaatg cagagcttgc aattcgcgct ttttcaatat
     7621 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7681 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7741 gaaaccatta ttatcatgac attaacctat aaaaataggc gtattacgag gccctttcac
     7801 g
//
