LOCUS       sequence-346597-        5983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of zebrafish EGFP-Bin1b. Parton lab clone HYG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5983)
  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 5983)
  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-Bin1b
FEATURES             Location/Qualifiers
     source          1..5983
                     /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(1975..1995)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    2035..2169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2059..2078)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2113..2132
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2282..2302)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2282..2300)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2321..2337)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2321..2337)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2334..2356)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2345..2361
                     /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(2369..2375,2376..2393,2394..2399))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2414..2435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2552..2569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2723..3311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2803..2822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3482..4273,4274..4342))
                     /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     4105..4124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4343..4447)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4473..4928)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4610..4631)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4822..4841
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4999..5983
                     /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 tggcagaggt tgggaaagga
      901 gtcaacgccg ggaaactggc cagtaatgta cagaagaggt taacccgagc acaagagaag
      961 gtgatgcaga aactgggaaa ggcggatgaa accaaagaca cagcctttga ggagcaagta
     1021 gccaacttta acaaacagat gaatgagggc accaaactgc agaaagactt gaaagcgtat
     1081 ttgctggcag tcaaaacaat gcacgagtgt tccaagcgtc tgcacgactg tctggcagaa
     1141 atgtatgacc ctgagtggtt tggcaaagag gaggtggact ccatcgcaga ggacactgac
     1201 ctcctgtggc aagatttcca cgagaagctt ttggacaatg ctgtaatttc catggacgca
     1261 tacttggctc agtttccaga tataaaggct cgcattgcaa agcgtgaaag gaagctggtg
     1321 gactttgaca gtgccagaca tcattttgcc tctgtacaaa aaggcaagaa aaaggacgag
     1381 gccaaaattg ccaaggctga ggaacacttg ggcagagctc agaaagtgtt tgaggagatc
     1441 aattatgatc tccaggagga acttcccacc ctgtggaaca gtcgtattgg cctctatgtt
     1501 aacacattcc ggagtgtgtc tggcctggag gaaacatttc acagggacat gggcaaactg
     1561 aaccagaatc tgagtgatat tatgaccaaa ctggaagagc agcgtgttgc caaaaaaggt
     1621 gtcacgacag caagcactgg gaagagtgac gatgcagcaa accacaatca ggagacgtca
     1681 gatgcaccta aatctccatc aaagaatgaa gccccaacgg tgacaaatgg ctcctcagat
     1741 acagagcttc ctccatctgt tctctttaag gtgaaaacaa tgcacgacta cagtgcaaca
     1801 gacagtgatg aacttgacct gaaggctggt gacattgtgc ttgtgcttcc ctttgccaac
     1861 ccagatgaag cggatgatgg ctggttaatg ggtgtaaaag agtcccactg gcttcagaac
     1921 aaaaacctcc tagcaaaagg agtcttccct gaaaacttca cccagaaact gtgacaactt
     1981 tattatacat agttgcactc gagcctctag aactatagtg agtcgtatta cgtagatcca
     2041 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2101 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2161 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2221 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     2281 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2341 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     2401 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2461 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2521 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2581 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2641 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2701 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2761 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2821 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2881 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2941 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3001 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3061 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3121 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3181 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3241 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3301 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3361 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3421 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3481 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3541 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3601 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3661 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3721 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3781 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3841 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3901 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3961 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4021 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4081 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4141 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4201 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4261 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4321 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4381 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4441 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     4501 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4561 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4621 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4681 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4741 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4801 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4861 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4921 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4981 ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa
     5041 ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa
     5101 ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa
     5161 ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca
     5221 agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca
     5281 ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc
     5341 cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     5401 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     5461 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     5521 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     5581 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     5641 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccgttctca
     5701 cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt
     5761 aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc
     5821 ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg
     5881 tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg
     5941 tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
