LOCUS       sequence-346605-        5846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of zebrafish Cavin4b-EGFP. Parton lab clone 
            HXM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5846)
  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 5846)
  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: Cavin4b-EGFP
FEATURES             Location/Qualifiers
     source          1..5846
                     /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(2148..2172)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2174..2176,2177..2890)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2174..2176,2177..2887)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2216..2237)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2477..2496)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2824..2845
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2892..2912)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    2952..3086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2976..2995)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3030..3049
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3199..3219)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3199..3217)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3238..3254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3238..3254)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3251..3273)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3262..3278
                     /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(3286..3292,3293..3310,3311..3316))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3331..3352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3469..3486)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3640..4228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3720..3739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4399..5190,5191..5259))
                     /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     5022..5041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5260..5364)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5390..5845)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5527..5548)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5739..5758
                     /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 ccatggctga
     1201 taaactggga ctggcaggtg tggcggacga gcccagcgcc atgaacatct tctctctgct
     1261 ggagagggtg tccgggatca tagacaatgt tcaggcctgc cagcagcgga tggaggagag
     1321 gcagctggag ctggagaaca ccgtcaagac catccaagca gatgtagtca agctgacgaa
     1381 ggagcacacg gcgaccagtg gcaccgtgga gaaacttctg gagaaaacac gcaaggtcag
     1441 ctgccacatc aaggacgtcc gcgtgcgggt ggagaagcag aacgttcgag tcaagaaggt
     1501 ggaggaaacc caagtcgagc tgctgaataa gaacaagttc cgtgtcgtca tctaccaggg
     1561 agataatgag gtcccagctg tggctggtcc taaagcctct ccgaagggag caatgggtgg
     1621 agacgctgct gcagacccag atgcaccaga ggctccgctc cctgactcgg atgaggaata
     1681 catggttgtt gaagaagccg attcttccac tgctgccaaa ctaaagaaga ccggcctgaa
     1741 gcgcatcgaa agcctgaagc agaccttctc acgcgagaac atgactaaga ccaaagaaaa
     1801 cctgggcacc aaggtcaaca agctgggcga gcgcatagta acagccgaac ggcgtgagaa
     1861 aatccgccag tcaggagaac gactgaaaca gtctggcgag cgcttcaagg aaaccatcac
     1921 aaaaaccgta ccagcaaagc tgaatctgaa gaaggaaagg acggtggctg agggtcagga
     1981 gggggcagaa gggaccacag aaggaacagc acccgtccca ccaccgaagg gtcgtaaacc
     2041 cagtcctgag gtggcctaca cagagccaac gcagaaacaa gaaggtccag atgagggcaa
     2101 gggtgaggaa tcagaagtac cgatgtatga tatgaagcag ctttctgacc cagctttctt
     2161 gtacaaagtg gctgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2221 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2281 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2341 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2401 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2461 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2521 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2581 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2641 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2701 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2761 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2821 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2881 gtacaagtaa ccaactttat tatacatagt tgcactcgag cctctagaac tatagtgagt
     2941 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta
     3001 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3061 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3121 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc
     3181 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3241 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3301 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3481 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3541 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4081 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4201 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4261 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4321 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4381 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4441 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4501 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4561 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4621 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4681 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4741 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4801 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4861 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4921 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4981 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5041 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5101 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5161 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5221 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5281 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5341 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     5401 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5461 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5521 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5581 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5641 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5701 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5761 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5821 cttaatgcgc cgctacaggg cgcgtc
//
