LOCUS       sequence-205192-        5640 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 5640)
  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 5640)
  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..5640
                     /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(1582..1681)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1689..1823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1713..1732)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1767..1786
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1936..1956)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1936..1954)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1975..1991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1975..1991)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1988..2010)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1999..2015
                     /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(2023..2029,2030..2047,2048..2053))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2068..2089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2206..2223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2377..2965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2457..2476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3136..3927,3928..3996))
                     /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     3759..3778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3997..4101)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4127..4582)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4264..4285)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4476..4495
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4656..5640
                     /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 atatcgagga tccgacgctt
      421 tatatagtag aacggccgtt gccggggtat ccggacgctg aggctcccga gccaagctca
      481 gcgggagcgc aagcagccga agaacccagt ggtgcaggca gcgaggagct catcaagagc
      541 gaccaagtca atggggtttt ggttctgagt ctcctcgata agataatagg ggccgtcgat
      601 cagattcagt tgacacaggc acaactcgaa gagcggcagg cggagatgga aggagcagtg
      661 caatctattc aaggcgagtt gtctaagctg ggcaaagcgc atgctacaac ttccaacact
      721 gtaagtaagc tgcttgaaaa ggtacgaaaa gtgtcagtta acgtaaaaac ggttcgcgga
      781 tctctcgaaa ggcaggccgg acagataaag aagctcgaag tgaacgaagc tgagctgttg
      841 cggcggcgca acttcaaagt gatgatatac caggatgagg tcaagctccc tgcgaaactg
      901 tccataagca aatcccttaa agagagcgag gcacttcccg agaaggaggg agaagagctt
      961 ggcgaaggtg agagacccga ggaggatgcg gcagccctcg agctcagtag tgatgaagcc
     1021 gtagaagtcg aggaagtcat cgaagagagt agggcagaaa ggataaaacg ctcaggcctg
     1081 cgccgcgttg acgatttcaa gaaagcgttc tccaaggaaa aaatggaaaa gacaaaagtt
     1141 aggactagag agaatctgga aaagacccga cttaagacca aagaaaacct ggagaaaact
     1201 aggcatactc tggagaaacg aatgaataag ctcggcacgc gattggtacc agccgaacgt
     1261 cgcgaaaagc tgaaaaccag tagggataag ctccgcaaga gcttcactcc ggaccatgtt
     1321 gtctatgcaa gatccaagac agcagtatat aaggtacccc ctttcacatt tcatgtcaaa
     1381 aagattagag aggggcaagt ggaagtcctc aaggccactg aaatggtcga ggtgggtgcc
     1441 gacgacgacg aagggggtgc ggagcgagga gaagctggtg atcttcgacg gggcagctca
     1501 cctgacgtgc atgcgttgct ggaaataacg gaagagagtg acgcggttct ggtggacaag
     1561 agcgatagcg actgaactag tacccagctt tcttgtacaa agttggcatt ataagaaagc
     1621 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1681 gatacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1741 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1801 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1861 agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt
     1921 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     1981 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2041 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2101 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2161 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     4141 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4201 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4261 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4321 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4381 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4441 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4501 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4561 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4621 cgatcggtgc gggcctcttc gctattacgc cagtcgacca tagccaattc aatatggcgt
     4681 atatggactc atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt
     4741 atatggactc gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg
     4801 acttggcacc atgccaattc aatatggcgg acttggcact gtgccaactg gggaggggtc
     4861 tacttggcac ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt
     4921 gaattggcat ggtgccaata atggcggcca tattggctat atgccaggat caatatatag
     4981 gcaatatcca atatggccct atgccaatat ggctattggc caggttcaat actatgtatt
     5041 ggccctatgc catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc
     5101 caatgggcgg tcccatatac catatatggg gcttcctaat accgcccata gccactcccc
     5161 cattgacgtc aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta
     5221 ttgacgtata tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca
     5281 atgcccattg acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc
     5341 attcatatcc gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca
     5401 gtacatcaat atctattaat agtaacttgg caagtacatt actattggaa ggacgccagg
     5461 gtacattggc agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa
     5521 gtacatcccc attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg
     5581 taaatgggcg gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
