LOCUS       sequence-209950-        5637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cavin1 fused to the C-terminus of mVenus for 
            bimolecular fluorescence complementation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5637)
  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 5637)
  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: mVenusC-Cavin1
FEATURES             Location/Qualifiers
     source          1..5637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..55
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     37..54
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      105..114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             114..365
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     302..323
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1584..1683)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1691..1825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1715..1734)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1769..1788
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1938..1958)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1938..1956)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1977..1993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1977..1993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1990..2012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2001..2017
                     /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(2025..2031,2032..2049,2050..2055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2070..2091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2208..2225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2379..2967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2459..2478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3138..3929,3930..3998))
                     /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     3761..3780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3999..4103)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4129..4584)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4266..4287)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4478..4497
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4655..5637,1..2)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca
       61 agctacttgt tctttttgca ggatcccatc gattcgaatt cgccgccacc atggacaagc
      121 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
      181 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      241 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      301 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      361 acaagggcgg aggaagcgga ggcggaagcg gcggaggatc tgatatcgag gatccgacgc
      421 tttatatagt agaacggccg ttgccggggt atccggacgc tgaggctccc gagccaagct
      481 cagcgggagc gcaagcagcc gaagaaccca gtggtgcagg cagcgaggag ctcatcaaga
      541 gcgaccaagt caatggggtt ttggttctga gtctcctcga taagataata ggggccgtcg
      601 atcagattca gttgacacag gcacaactcg aagagcggca ggcggagatg gaaggagcag
      661 tgcaatctat tcaaggcgag ttgtctaagc tgggcaaagc gcatgctaca acttccaaca
      721 ctgtaagtaa gctgcttgaa aaggtacgaa aagtgtcagt taacgtaaaa acggttcgcg
      781 gatctctcga aaggcaggcc ggacagataa agaagctcga agtgaacgaa gctgagctgt
      841 tgcggcggcg caacttcaaa gtgatgatat accaggatga ggtcaagctc cctgcgaaac
      901 tgtccataag caaatccctt aaagagagcg aggcacttcc cgagaaggag ggagaagagc
      961 ttggcgaagg tgagagaccc gaggaggatg cggcagccct cgagctcagt agtgatgaag
     1021 ccgtagaagt cgaggaagtc atcgaagaga gtagggcaga aaggataaaa cgctcaggcc
     1081 tgcgccgcgt tgacgatttc aagaaagcgt tctccaagga aaaaatggaa aagacaaaag
     1141 ttaggactag agagaatctg gaaaagaccc gacttaagac caaagaaaac ctggagaaaa
     1201 ctaggcatac tctggagaaa cgaatgaata agctcggcac gcgattggta ccagccgaac
     1261 gtcgcgaaaa gctgaaaacc agtagggata agctccgcaa gagcttcact ccggaccatg
     1321 ttgtctatgc aagatccaag acagcagtat ataaggtacc ccctttcaca tttcatgtca
     1381 aaaagattag agaggggcaa gtggaagtcc tcaaggccac tgaaatggtc gaggtgggtg
     1441 ccgacgacga cgaagggggt gcggagcgag gagaagctgg tgatcttcga cggggcagct
     1501 cacctgacgt gcatgcgttg ctggaaataa cggaagagag tgacgcggtt ctggtggaca
     1561 agagcgatag cgactgaact agtacccagc tttcttgtac aaagttggca ttataagaaa
     1621 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1681 ttgatacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1741 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1801 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1861 tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg
     1921 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     1981 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg
     2041 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2101 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2161 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2221 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2281 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2341 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2401 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2461 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2521 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2581 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2641 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2701 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2761 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2821 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2881 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2941 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3001 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3061 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3121 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3181 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3241 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3301 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3361 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3421 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3481 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3541 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3601 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3661 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3721 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3781 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3841 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3901 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3961 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4021 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4081 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     4141 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4201 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4261 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4321 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4381 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4441 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4501 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4561 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     4621 ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta
     4681 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
     4741 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
     4801 cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct
     4861 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
     4921 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
     4981 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
     5041 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
     5101 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
     5161 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
     5221 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
     5281 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
     5341 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
     5401 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg
     5461 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
     5521 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
     5581 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttaggga
//
