LOCUS       sequence-229456-        6863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bimolecular Fluorescence Complimentation (BiFC) assay. Mammalian 
            expression vector to generate stable, inducible cell line expressing
            N-terminally tagged HA-GFP1-10-actin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6863)
  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 6863)
  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-actin-pcDNA4-TO
FEATURES             Location/Qualifiers
     source          1..6863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1007..1033
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2871..2888)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2877..3101
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3147..3575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3234..3253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3444..3465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3584..3604)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3589..3918
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3769..3904
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3831..3850
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3966..4013
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4032..4406
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4536..4657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4573..4592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4627..4646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4706..4722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4706..4722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4719..4741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4730..4746
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4754..4760,4761..4778,4779..4784))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4799..4820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4937..4954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5108..5696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5188..5207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5867..6658,6659..6727))
                     /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     6490..6509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6728..6832)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcgggcc accatgtacc catacgatgt
     1021 tccagattac gctgtttcga aaggcgagga gctgttcaca ggcgtggtgc caatcctggt
     1081 ggagctggac ggcgacgtga acggccacaa attcagcgtg agaggcgagg gcgagggcga
     1141 cgccacaatc ggcaaactga cactgaaatt catctgcaca acaggcaaac tgccagtgcc
     1201 ctggccaaca ctagtgacaa cactgacata cggcgtgcag tgcttcagca gatatccgga
     1261 ccacatgaaa agacacgact tcttcaaaag cgccatgcca gagggctacg tgcaggagag
     1321 aacaatcagc ttcaaagacg acggcaaata caaaacaaga gccgtggtga aattcgaggg
     1381 cgacacactg gtgaacagaa tcgagctgaa aggcacagac ttcaaagagg acggcaacat
     1441 cctgggccac aaactggagt acaacttcaa cagccacaac gtgtacatca cagccaacaa
     1501 acagaaaaac ggcatcaaag ccaacttcac agtgagacac aacgtggagg acggcagcgt
     1561 gcagctcgcc gaccactacc agcagaacac accaatcggc gacggcccag tgctgctgcc
     1621 agacaaccac tacctgagca cacagacagt gctgagcaaa gacccaaacg agaaatccgg
     1681 actcagatct cgagctcaag cttctatgga tgatgatatc gccgcgctcg tcgtcgacaa
     1741 cggctccggc atgtgcaagg ccggcttcgc gggcgacgat gccccccggg ccgtcttccc
     1801 ctccatcgtg gggcgcccca ggcaccaggg cgtgatggtg ggcatgggtc agaaggattc
     1861 ctatgtgggc gacgaggccc agagcaagag aggcatcctc accctgaagt accccatcga
     1921 gcacggcatc gtcaccaact gggacgacat ggagaaaatc tggcaccaca ccttctacaa
     1981 tgagctgcgt gtggctcccg aggagcaccc cgtgctgctg accgaggccc ccctgaaccc
     2041 caaggccaac cgcgagaaga tgacccagat catgtttgag accttcaaca ccccagccat
     2101 gtacgttgct atccaggctg tgctatccct gtacgcctct ggccgtacca ctggcatcgt
     2161 gatggactcc ggtgacgggg tcacccacac tgtgcccatc tacgaggggt atgccctccc
     2221 ccatgccatc ctgcgtctgg acctggctgg ccgggacctg actgactacc tcatgaagat
     2281 cctcaccgag cgcggctaca gcttcaccac cacggccgag cgggaaatcg tgcgtgacat
     2341 taaggagaag ctgtgctacg tcgccctgga cttcgagcaa gagatggcca cggctgcttc
     2401 cagctcctcc ctggagaaga gctacgagct gcctgacggc caggtcatca ccattggcaa
     2461 tgagcggttc cgctgccctg aggcactctt ccagccttcc ttcctgggca tggagtcctg
     2521 tggcatccac gaaactacct tcaactccat catgaagtgt gacgtggaca tccgcaaaga
     2581 cctgtacgcc aacacagtgc tgtctggcgg caccaccatg taccctggca ttgccgacag
     2641 gatgcagaag gagatcactg ccctggcacc cagcacaatg aagatcaaga tcattgctcc
     2701 tcctgagcgc aagtactccg tgtggatcgg cggctccatc ctggcctcgc tgtccacctt
     2761 ccagcagatg tggatcagca agcaggagta tgacgagtcc ggcccctcca tcgtccaccg
     2821 caaatgcttc taactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     2881 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2941 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3001 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3061 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3121 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3301 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3361 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3481 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3541 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3601 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3721 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3781 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3841 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3901 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     3961 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     4021 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     4081 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     4141 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     4201 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     4261 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     4321 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     4381 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     4441 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4501 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4561 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4621 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4681 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4741 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4801 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4861 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5521 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5761 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5821 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5881 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5941 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6001 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6061 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6121 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6181 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6241 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6301 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6361 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6421 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6481 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6541 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6601 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6661 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6721 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6781 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6841 cccgaaaagt gccacctgac gtc
//
