LOCUS       sequence-346601-        6337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of zebrafish EGFP-Bin1a. Parton lab clone HYA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6337)
  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 6337)
  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: EGFP-Bin1a
FEATURES             Location/Qualifiers
     source          1..6337
                     /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"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      133..142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(139..141,142..144,145..855)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(139..141,142..144,145..855)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(184..205)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(445..464)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     792..813
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(857..881)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2329..2349)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    2389..2523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2413..2432)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2467..2486
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2636..2656)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2636..2654)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2675..2691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2675..2691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2688..2710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2699..2715
                     /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(2723..2729,2730..2747,2748..2753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2768..2789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2906..2923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3077..3665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3157..3176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3836..4627,4628..4696))
                     /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     4459..4478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4697..4801)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4827..5282)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4964..4985)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5176..5195
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5353..6337
                     /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 ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg ctgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      181 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      241 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      301 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      361 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      421 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      481 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      541 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      601 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      661 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      721 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
      781 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      841 gacgagctgt acaagcaccc agctttcttg tacaaagtgg tggctgaatt accgatgggg
      901 aaaggggtgt ccgctggaaa aatagcaagc aatgtacaga agaaaattac tagagcacag
      961 gagaaggtta tgcaaaagct tggaaaggca gatgagacta aagatctggc gtttgaagag
     1021 ggcgtgatca actttaataa acagttggtg gaaggtacca aactgcagaa agaccttcgg
     1081 gcctatttga ctgctgtgaa agccatgcac gagtcttcta gacgtctgta cgagtgcctg
     1141 gatgacatgt atgagacaga gtggtacggc agaaagaaaa tggacgcagt tgtagaggat
     1201 tcagacctgc tttggacaga cttccatcag aaactcgtgg atcatgcgct tatatccatg
     1261 gacacttacc tgggccagtt tccagacatc aaagctcgca ttgcgaaacg tgacagaaag
     1321 ctagttgact ttgacagcgc acggcacaac tttgctgctg taaacaaagg caagaagaaa
     1381 gaaggggtca agatcaccaa gccagcctct ttgctggaaa gggctgcccc aagctgggct
     1441 caggggatcc tgtccgccca taatgtggct caaaccagcc tctccaggag tcaggcagag
     1501 gaggagctgg agagggcaca gaaagtcttt gaggaaatta acatagattt gcaggaggaa
     1561 ctgccatctc tgtggaacag tcgtgttggc ttttatgtga atacgttcca gagtgtggca
     1621 ggtctggagg aaaagtttca cagagaaatg ggaaaaatta atcagaatct ttacgataca
     1681 ctggacaaac tggaaaatca agatatgacc agtgaaagaa cacaagatgc caatcacact
     1741 ttaagcccag gaggaccacc agccattccc aaatcgccgt ctaagctaag gcctgcagtt
     1801 cctcctcctc ccaaagtaac tccgtcaaag gacctgaagc aggagagcat catcaacctg
     1861 tttgaagagg ccgccactcc aaacatcagt gtcacttcac caacagagta tgatgctcca
     1921 gccactggta atcttctgga tatggatctg gactcacttc aagctgccac cacaccagct
     1981 gcacagacgt gggattcatg ggagactgca gaaacaacag ctgttacaga agaggcagca
     2041 gcagctgaac ctgtaagtga gactgtgcag gagagcgccg agagcccaga gagcgtagag
     2101 atgccagaat ccttcctgtt caaagtgaaa gctatgcacg actacaatgc caccgactct
     2161 gatgagctgg agttaaaggc gggcgacgtg gtgctggtag tgaattatga aaatccagac
     2221 gatcaggatg acggctggtt gatgggagtc aaagaggctg attggatcca gcagaaagat
     2281 cttgggaaaa aaggcgtgtt ccctgaaaac ttcacccaga ggatgtgaca actttattat
     2341 acatagttgc actcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg
     2401 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2461 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2521 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2581 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     2641 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2701 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
     2761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4801 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     4861 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4921 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4981 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5041 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5101 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5161 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5221 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5281 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5341 gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat
     5401 atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat
     5461 atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat
     5521 atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg
     5581 aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg
     5641 gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg
     5701 ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat atagtattcc
     5761 atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat
     5821 atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata
     5881 tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc
     5941 attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac
     6001 cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc
     6061 ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt
     6121 aacttggcaa gtacattact attggaagta cgccagggta cattggcagt actcccattg
     6181 acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg
     6241 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
     6301 atgggaggtc tatataagca atgctcgttt agggaac
//
