LOCUS       sequence-210166-        5460 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 5460)
  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 5460)
  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..5460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..49
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     31..48
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(105..107,108..110,111..569)
                     /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="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDVTYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(150..171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(411..430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1401..1500)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1508..1642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1532..1551)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1586..1605
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1755..1775)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1755..1773)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1794..1810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1794..1810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1807..1829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1818..1834
                     /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(1842..1848,1849..1866,1867..1872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1887..1908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2025..2042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2196..2784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2276..2295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2955..3746,3747..3815))
                     /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     3578..3597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3816..3920)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3946..4401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4083..4104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4295..4314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4472..5456
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 attctgcctg gggacgtcgg agcaagcttg atttaggtga cactatagaa tacaagctac
       61 ttgttctttt tgcaggatcc catcgattcg aattcgccgc caccatggtg agcaagggcg
      121 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      181 acaagttcag cgtgtccggc gagggcgagg gcgatgtcac ctacggcaag ctgaccctga
      241 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
      301 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      361 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      421 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      481 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      541 acaacagcca caacgtctat atcaccgccg gcggaggaag cggaggcgga agcggcggag
      601 gatctgatat ccgagagagc gccttggaga gggggcccgt gccagaggca cccgccggtg
      661 ggccagttca cgcagtgaca gtggttacgc ttctggaaaa actggccagt atgctcgaaa
      721 cacttagaga gcgccagggg ggcctggccc ggaggcaagg cggcctggca ggatctgtaa
      781 ggagaatcca gagcggcctg ggcgccctgt caaggtcaca cgacaccacc agcaacaccc
      841 tggcccagct gttggccaag gctgagcgag tttcttctca cgcaaatgct gcacaggaac
      901 gagctgtgag aagggctgcc caggtccagc ggctggaggc caatcacggc ctgttggtcg
      961 ccagaggcaa gttgcacgtg ctgctgttca aagaggaagg agaggtgcct gccagcgcct
     1021 tccagaaagc tccggaaccg ttgggtccgg cagatcagtc agaactgggg cccgagcagt
     1081 tggaggccga agtgggcgaa tcatcagatg aggagccggt tgaatcacgc gctcaaaggt
     1141 tgcggagaac cggcctgcaa aaggtgcagt cattgcgccg cgcactttct ggaagaaagg
     1201 gcccagcagc cccacccccc actccagtta aaccaccccg gctgggaccc gggcggtcag
     1261 ctgaggctca gcccgaggct cagcctgccc tggagccaac cctggaaccc gagccaccgc
     1321 aagatacaga ggaggatcca ggccggcctg gcgccgccga agaggctctt ctccagatgg
     1381 agtctgtggc ctgaactagt acccagcttt cttgtacaaa gttggcatta taagaaagca
     1441 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1501 atacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1561 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1621 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1681 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     1741 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     1801 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc
     1861 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1921 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1981 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2041 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2101 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2281 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2341 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2401 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2461 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2521 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2581 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2641 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2701 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2761 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2821 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2881 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2941 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3001 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3061 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3121 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3181 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3241 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3301 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3361 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3421 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3481 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3541 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3601 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3661 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3721 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3781 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3841 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3901 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     3961 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4021 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4081 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4141 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4201 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4261 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4321 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4381 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     4441 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat
     4501 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
     4561 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
     4621 ggcaccatgc caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact
     4681 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
     4741 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
     4801 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
     4861 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
     4921 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
     4981 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
     5041 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
     5101 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
     5161 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
     5221 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac
     5281 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac
     5341 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa
     5401 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaaccgcc
//
