LOCUS       sequence-205193-        5850 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 5850)
  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 5850)
  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..5850
                     /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(1792..1891)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1899..2033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1923..1942)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1977..1996
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2146..2166)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2146..2164)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2185..2201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2185..2201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2198..2220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2209..2225
                     /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(2233..2239,2240..2257,2258..2263))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2278..2299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2416..2433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2587..3175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2667..2686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3346..4137,4138..4206))
                     /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     3969..3988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4207..4311)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4337..4792)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4474..4495)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4686..4705
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4866..5850
                     /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 atatcgagga tccgacgctt tatatagtag aacggccgtt gccggggtat
      661 ccggacgctg aggctcccga gccaagctca gcgggagcgc aagcagccga agaacccagt
      721 ggtgcaggca gcgaggagct catcaagagc gaccaagtca atggggtttt ggttctgagt
      781 ctcctcgata agataatagg ggccgtcgat cagattcagt tgacacaggc acaactcgaa
      841 gagcggcagg cggagatgga aggagcagtg caatctattc aaggcgagtt gtctaagctg
      901 ggcaaagcgc atgctacaac ttccaacact gtaagtaagc tgcttgaaaa ggtacgaaaa
      961 gtgtcagtta acgtaaaaac ggttcgcgga tctctcgaaa ggcaggccgg acagataaag
     1021 aagctcgaag tgaacgaagc tgagctgttg cggcggcgca acttcaaagt gatgatatac
     1081 caggatgagg tcaagctccc tgcgaaactg tccataagca aatcccttaa agagagcgag
     1141 gcacttcccg agaaggaggg agaagagctt ggcgaaggtg agagacccga ggaggatgcg
     1201 gcagccctcg agctcagtag tgatgaagcc gtagaagtcg aggaagtcat cgaagagagt
     1261 agggcagaaa ggataaaacg ctcaggcctg cgccgcgttg acgatttcaa gaaagcgttc
     1321 tccaaggaaa aaatggaaaa gacaaaagtt aggactagag agaatctgga aaagacccga
     1381 cttaagacca aagaaaacct ggagaaaact aggcatactc tggagaaacg aatgaataag
     1441 ctcggcacgc gattggtacc agccgaacgt cgcgaaaagc tgaaaaccag tagggataag
     1501 ctccgcaaga gcttcactcc ggaccatgtt gtctatgcaa gatccaagac agcagtatat
     1561 aaggtacccc ctttcacatt tcatgtcaaa aagattagag aggggcaagt ggaagtcctc
     1621 aaggccactg aaatggtcga ggtgggtgcc gacgacgacg aagggggtgc ggagcgagga
     1681 gaagctggtg atcttcgacg gggcagctca cctgacgtgc atgcgttgct ggaaataacg
     1741 gaagagagtg acgcggttct ggtggacaag agcgatagcg actgaactag tacccagctt
     1801 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     1861 tcactatcag tcaaaataaa atcattattt gatacgtaga tccagacatg ataagataca
     1921 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1981 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2041 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     2101 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     2161 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2221 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2281 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2341 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2761 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3001 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3241 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3301 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3361 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3421 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3481 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3541 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3601 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3661 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3721 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3781 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3841 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3901 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3961 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4021 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4081 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4141 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4201 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4261 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4321 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4381 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4441 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4501 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4561 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4621 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4681 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4741 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     4801 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4861 cagtcgacca tagccaattc aatatggcgt atatggactc atgccaattc aatatggtgg
     4921 atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg
     4981 atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg
     5041 acttggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg
     5101 gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca
     5161 tattggctat atgccaggat caatatatag gcaatatcca atatggccct atgccaatat
     5221 ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg
     5281 ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg
     5341 gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc
     5401 tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg
     5461 tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac
     5521 cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga
     5581 cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg
     5641 caagtacatt actattggaa ggacgccagg gtacattggc agtactccca ttgacgtcaa
     5701 tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg
     5761 gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag
     5821 gtctatataa gcaatgctcg tttagggaac
//
