LOCUS       sequence-205190-        5253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cavin3 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 5253)
  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 5253)
  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-Cavin3
FEATURES             Location/Qualifiers
     source          1..5253
                     /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             112..363
                     /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     300..321
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1195..1294)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1302..1436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1326..1345)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1380..1399
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1549..1569)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1549..1567)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1588..1604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1588..1604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1601..1623)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1612..1628
                     /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(1636..1642,1643..1660,1661..1666))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1681..1702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1819..1836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1990..2578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2070..2089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2749..3540,3541..3609))
                     /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     3372..3391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3610..3714)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3740..4195)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3877..3898)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4089..4108
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4269..5253
                     /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 ggacaagcag
      121 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
      181 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      241 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
      301 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      361 aagggcggag gaagcggagg cggaagcggc ggcggctctg atatccgaga gagcgccttg
      421 gagagggggc ccgtgccaga ggcacccgcc ggtgggccag ttcacgcagt gacagtggtt
      481 acgcttctgg aaaaactggc cagtatgctc gaaacactta gagagcgcca ggggggcctg
      541 gcccggaggc aaggcggcct ggcaggatct gtaaggagaa tccagagcgg cctgggcgcc
      601 ctgtcaaggt cacacgacac caccagcaac accctggccc agctgttggc caaggctgag
      661 cgagtttctt ctcacgcaaa tgctgcacag gaacgagctg tgagaagggc tgcccaggtc
      721 cagcggctgg aggccaatca cggcctgttg gtcgccagag gcaagttgca cgtgctgctg
      781 ttcaaagagg aaggagaggt gcctgccagc gccttccaga aagctccgga accgttgggt
      841 ccggcagatc agtcagaact ggggcccgag cagttggagg ccgaagtggg cgaatcatca
      901 gatgaggagc cggttgaatc acgcgctcaa aggttgcgga gaaccggcct gcaaaaggtg
      961 cagtcattgc gccgcgcact ttctggaaga aagggcccag cagccccacc ccccactcca
     1021 gttaaaccac cccggctggg acccgggcgg tcagctgagg ctcagcccga ggctcagcct
     1081 gccctggagc caaccctgga acccgagcca ccgcaagata cagaggagga tccaggccgg
     1141 cctggcgccg ccgaagaggc tcttctccag atggagtctg tggcctgaac tagtacccag
     1201 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     1261 aggtcactat cagtcaaaat aaaatcatta tttgatacgt agatccagac atgataagat
     1321 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1381 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1441 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     1501 tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg
     1561 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1621 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1681 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1741 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1801 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1861 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1921 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1981 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2041 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2101 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2161 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2221 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2281 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2341 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2401 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     2461 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2521 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2581 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2641 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2701 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2761 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2821 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2881 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2941 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3001 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3061 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3121 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3181 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3241 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3301 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3361 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3421 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3481 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3541 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3601 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3661 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3721 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3781 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3841 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3901 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3961 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4021 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4081 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4141 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     4201 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4261 cgccagtcga ccatagccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
     4321 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
     4381 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
     4441 cggacttggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
     4501 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
     4561 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
     4621 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
     4681 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
     4741 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
     4801 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
     4861 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
     4921 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
     4981 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
     5041 tggcaagtac attactattg gaaggacgcc agggtacatt ggcagtactc ccattgacgt
     5101 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
     5161 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
     5221 gaggtctata taagcaatgc tcgtttaggg aac
//
