LOCUS       sequence-205191-        5463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cavin3 fused to the N-terminus of mVenus for 
            bimolecular fluorescence complementation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5463)
  AUTHORS   Ariotti N, Rae J, Giles N, Martel N, Sierecki E, Gambin Y, Hall TE, 
            Parton RG
  TITLE     Ultrastructural localisation of protein interactions using 
            conditionally stable nanobodies.
  JOURNAL   PLoS Biol. 2018 Apr 5;16(4):e2005473. doi: 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr.
  PUBMED    29621251
REFERENCE   2  (bases 1 to 5463)
  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: "PLoS 
            Biol."; date: "2018-04-5"; volume: "16(4):e2005473. doi"; pages: " 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr"
COMMENT     Sequence Label: mVenusN-Cavin3
FEATURES             Location/Qualifiers
     source          1..5463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      103..112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(109..111,112..114,115..573)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(154..175)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(415..434)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1405..1504)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1512..1646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1536..1555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1590..1609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1759..1779)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1759..1777)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1798..1814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1798..1814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1811..1833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1822..1838
                     /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(1846..1852,1853..1870,1871..1876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1891..1912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2029..2046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2200..2788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2280..2299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2959..3750,3751..3819))
                     /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     3582..3601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3820..3924)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3950..4405)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4087..4108)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4299..4318
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4479..5463
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg ccgccaccat ggtgagcaag
      121 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      181 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      241 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      301 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      361 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      421 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      481 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      541 aactacaaca gccacaacgt ctatatcacc gccggcggag gaagcggagg cggaagcggc
      601 ggcggctctg atatccgaga gagcgccttg gagagggggc ccgtgccaga ggcacccgcc
      661 ggtgggccag ttcacgcagt gacagtggtt acgcttctgg aaaaactggc cagtatgctc
      721 gaaacactta gagagcgcca ggggggcctg gcccggaggc aaggcggcct ggcaggatct
      781 gtaaggagaa tccagagcgg cctgggcgcc ctgtcaaggt cacacgacac caccagcaac
      841 accctggccc agctgttggc caaggctgag cgagtttctt ctcacgcaaa tgctgcacag
      901 gaacgagctg tgagaagggc tgcccaggtc cagcggctgg aggccaatca cggcctgttg
      961 gtcgccagag gcaagttgca cgtgctgctg ttcaaagagg aaggagaggt gcctgccagc
     1021 gccttccaga aagctccgga accgttgggt ccggcagatc agtcagaact ggggcccgag
     1081 cagttggagg ccgaagtggg cgaatcatca gatgaggagc cggttgaatc acgcgctcaa
     1141 aggttgcgga gaaccggcct gcaaaaggtg cagtcattgc gccgcgcact ttctggaaga
     1201 aagggcccag cagccccacc ccccactcca gttaaaccac cccggctggg acccgggcgg
     1261 tcagctgagg ctcagcccga ggctcagcct gccctggagc caaccctgga acccgagcca
     1321 ccgcaagata cagaggagga tccaggccgg cctggcgccg ccgaagaggc tcttctccag
     1381 atggagtctg tggcctgaac tagtacccag ctttcttgta caaagttggc attataagaa
     1441 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     1501 tttgatacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta
     1561 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1621 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1681 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc
     1741 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     1801 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1921 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2641 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     3961 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4021 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4081 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4141 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4201 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4261 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4321 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4381 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     4441 gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccatagccaa ttcaatatgg
     4501 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
     4561 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
     4621 cggacttggc accatgccaa ttcaatatgg cggacttggc actgtgccaa ctggggaggg
     4681 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
     4741 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
     4801 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
     4861 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
     4921 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
     4981 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
     5041 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
     5101 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
     5161 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
     5221 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaaggacgcc
     5281 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     5341 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     5401 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     5461 aac
//
