LOCUS       sequence-213324-        5847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cavin1 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 5847)
  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 5847)
  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-Cavin1
FEATURES             Location/Qualifiers
     source          1..5847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..1018
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1053..1071
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1053..1070
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1121..1130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1127..1129,1130..1132,1133..1591)
                     /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(1172..1193)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1433..1452)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2810..2909)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2917..3051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2941..2960)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2995..3014
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3164..3184)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3164..3182)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3203..3219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3203..3219)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3216..3238)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3227..3243
                     /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(3251..3257,3258..3275,3276..3281))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3296..3317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3434..3451)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3605..4193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3685..3704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4364..5155,5156..5224))
                     /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     4987..5006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5225..5329)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5355..5810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5492..5513)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5704..5723
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgatcggtg cgggcctctt cgctattacg ccagtcgacc gccaattcaa tatggcgtat
       61 atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat
      121 atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac
      181 ttggcaccat gccaattcaa tatggcggac ctggcactgt gccaactggg gaggggtcta
      241 cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga
      301 attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca atatataggc
      361 aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg
      421 ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca
      481 atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca
      541 ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt
      601 gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat
      661 gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat
      721 tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt
      781 acatcaatat ctattaatag taacttggca agtacattac tattggaagt acgccagggt
      841 acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt
      901 acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta
      961 aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaaccg
     1021 ccattctgcc tggggacgtc ggagcaagct tgatttaggt gacactatag aatacaagct
     1081 acttgttctt tttgcaggat cccatcgatt cgaattcgcc gccaccatgg tgagcaaggg
     1141 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1201 ccacaagttc agcgtgtccg gcgagggcga gggcgatgtc acctacggca agctgaccct
     1261 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1321 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1381 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1441 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1501 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1561 ctacaacagc cacaacgtct atatcaccgc cggcggagga agcggaggcg gaagcggcgg
     1621 cggctctgat atcgaggatc cgacgcttta tatagtagaa cggccgttgc cggggtatcc
     1681 ggacgctgag gctcccgagc caagctcagc gggagcgcaa gcagccgaag aacccagtgg
     1741 tgcaggcagc gaggagctca tcaagagcga ccaagtcaat ggggttttgg ttctgagtct
     1801 cctcgataag ataatagggg ccgtcgatca gattcagttg acacaggcac aactcgaaga
     1861 gcggcaggcg gagatggaag gagcagtgca atctattcaa ggcgagttgt ctaagctggg
     1921 caaagcgcat gctacaactt ccaacactgt aagtaagctg cttgaaaagg tacgaaaagt
     1981 gtcagttaac gtaaaaacgg ttcgcggatc tctcgaaagg caggccggac agataaagaa
     2041 gctcgaagtg aacgaagctg agctgttgcg gcggcgcaac ttcaaagtga tgatatacca
     2101 ggatgaggtc aagctccctg cgaaactgtc cataagcaaa tcccttaaag agagcgaggc
     2161 acttcccgag aaggagggag aagagcttgg cgaaggtgag agacccgagg aggatgcggc
     2221 agccctcgag ctcagtagtg atgaagccgt agaagtcgag gaagtcatcg aagagagtag
     2281 ggcagaaagg ataaaacgct caggcctgcg ccgcgttgac gatttcaaga aagcgttctc
     2341 caaggaaaaa atggaaaaga caaaagttag gactagagag aatctggaaa agacccgact
     2401 taagaccaaa gaaaacctgg agaaaactag gcatactctg gagaaacgaa tgaataagct
     2461 cggcacgcga ttggtaccag ccgaacgtcg cgaaaagctg aaaaccagta gggataagct
     2521 ccgcaagagc ttcactccgg accatgttgt ctatgcaaga tccaagacag cagtatataa
     2581 ggtaccccct ttcacatttc atgtcaaaaa gattagagag gggcaagtgg aagtcctcaa
     2641 ggccactgaa atggtcgagg tgggtgccga cgacgacgaa gggggtgcgg agcgaggaga
     2701 agctggtgat cttcgacggg gcagctcacc tgacgtgcat gcgttgctgg aaataacgga
     2761 agagagtgac gcggttctgg tggacaagag cgatagcgac tgaactagta cccagctttc
     2821 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     2881 actatcagtc aaaataaaat cattatttga tacgtagatc cagacatgat aagatacatt
     2941 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3001 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3061 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg
     3121 gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     3181 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3241 caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag
     3301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3421 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3481 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3541 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3781 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4021 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4141 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4201 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4261 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4321 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4381 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4441 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4501 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4561 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4621 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4681 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4741 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4801 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4861 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4921 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4981 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5041 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5101 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5161 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5221 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5281 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5341 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5401 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5461 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5521 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5581 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5641 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5701 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5761 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     5821 attcaggctg cgcaactgtt gggaagg
//
