LOCUS       sequence-346603-        5969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Mouse Cavin4-mCherry. Parton lab clone JSG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5969)
  AUTHORS   Lo HP, Lim YW, Xiong Z, Martel N, Ferguson C, Ariotti N, Giacomotto 
            J, Rae J, Floetenmeyer M, Moradi SV, Gao Y, Tillu VA, Xia D, Wang H,
            Rahnama S, Nixon SJ, Bastiani M, Day RD, Smith KA, Palpant NJ, 
            Johnston WA, Alexandrov K, Collins BM, Hall TE, Parton RG
  TITLE     Cavin4 interacts with Bin1 to promote T-tubule formation and 
            stability in developing skeletal muscle.
  JOURNAL   J Cell Biol. 2021 Dec 6;220(12). pii: 212693. doi: 
            10.1083/jcb.201905065. Epub 2021 Oct 11.
  PUBMED    34633413
REFERENCE   2  (bases 1 to 5969)
  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: "J Cell 
            Biol."; date: "2021-12-6"; volume: "220(12). pii: 212693. doi"; 
            pages: " 10.1083/jcb.201905065. Epub 2021 Oct 11"
COMMENT     Sequence Label: Cavin4-mCherry
FEATURES             Location/Qualifiers
     source          1..5969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..1054
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1089..1107
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1089..1106
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1162..1186
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1187..1196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2280..2304)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2306..3013
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2448..2466)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2716..2735
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2910..2933
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(3015..3035)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    3075..3209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3099..3118)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3153..3172
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3322..3342)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3322..3340)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3361..3377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3361..3377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3374..3396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3385..3401
                     /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(3409..3415,3416..3433,3434..3439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3454..3475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3592..3609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3763..4351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3843..3862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4522..5313,5314..5382))
                     /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     5145..5164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5383..5487)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5513..5968)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5650..5671)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5862..5881
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg
      121 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
      181 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
      241 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
      301 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
      361 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
      421 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
      481 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
      541 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
      601 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
      661 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
      721 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
      781 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
      841 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
      901 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
      961 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     1021 ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag
     1081 caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca
     1141 tcgattcgaa ttcaaggatc aacaagtttg tacaaaaaag caggctgcca ccatggaaca
     1201 caacggatca gcttcaaatg ctggtaaaat ccaccagaat cggttgtcaa gtgtgacgga
     1261 agatgaagac caagacgcag ccctcaccat tgtgaccgtg ctggacagag tggccagtgt
     1321 cgtggacagt gtgcaggcaa gccagaagag aattgaggag agacacagag aaatgggtaa
     1381 cgcgataaag tctgtccaaa tagacctgct gaagctctca cagtcacaca gcaatacggg
     1441 ctacgttgtc aacaagctgt ttgagaagac ccggaaagtc agcgctcaca ttaaggatgt
     1501 caaggcccgg gtagagaagc aacaggttcg tgtaaccaaa gtcgaaacca agcaagaaga
     1561 aataatgaag aaaaacaaat tccgagtggt aatcttccag gaggacattc cctgccccgc
     1621 atccctgtct gttgttaaag acagaagcct gcccgagaac caggaggagg ccgaagaagt
     1681 cttcgatccc ccaatagagc tctcctcgga tgaggagtat tatgttgaag aaagcagatc
     1741 tgccaggctt aggaagtcag gcaaagagca cattgaccac atcaagaaag ccttttccag
     1801 agaaaacatg cagaagacac ggcagactct tgacaagaaa gtgagtggga ttagaactag
     1861 gatagtcact cctgagagga gagagaggct gaggcagtcg ggagagaggc tgaggcagtc
     1921 aggagagagg ctgaggcagt cgggggaaag atttaagaaa tcgatttcca gtgccgctcc
     1981 ctcaaaggaa gcttttaaga ttcggagcct taggaaagcg aaggatccca aggccgaggg
     2041 ccaggaggta gacagaggga tgggcgtgga catcatctca ggtagcctgg ctctggggcc
     2101 catccatgag ttccactctg atgagttcag tgaaacagaa aaggaggtga ccaaaggagg
     2161 gtacagtccc caagaaggag gggacccccc aacgcctgag cctttgaagg tgacttttaa
     2221 accccaggtg agggtagagg atgacgagtc actcctgttg gagctaaagc agtcctcata
     2281 cccagctttc ttgtacaaag tggctgtgag caagggcgag gaggacaaca tggccatcat
     2341 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     2401 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2461 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     2521 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     2581 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     2641 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     2701 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     2761 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     2821 ggacggcggc cactacgacg ccgaggtcaa gaccacctac aaggccaaga agcccgtgca
     2881 gctgcccggc gcctacaacg tcaacatcaa gctggacatc acctcccaca acgaggacta
     2941 caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg gcatggacga
     3001 gctgtacaag taaccaactt tattatacat agttgcactc gagcctctag aactatagtg
     3061 agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     3121 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3181 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3241 aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg
     3301 cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     3361 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3421 cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4201 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4321 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4441 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4501 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4621 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4681 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4801 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5461 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     5521 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5581 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5641 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5701 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5761 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5821 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5881 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5941 gcgcttaatg cgccgctaca gggcgcgtc
//
