LOCUS       sequence-346600-        6253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human EGFP-Bin1 (transcriptional variant 8, 
            muscle specific). Parton lab clone HBH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6253)
  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 6253)
  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-HsBIN1_tv8
FEATURES             Location/Qualifiers
     source          1..6253
                     /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"
     CDS             join(1193..1195,1196..1198,1199..1909)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1193..1195,1196..1198,1199..1909)
                     /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(1238..1259)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1499..1518)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1846..1867
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1911..1935)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3299..3319)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    3359..3493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3383..3402)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3437..3456
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3606..3626)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3606..3624)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3645..3661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3645..3661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3658..3680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3669..3685
                     /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(3693..3699,3700..3717,3718..3723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3738..3759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3876..3893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4047..4635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4127..4146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4806..5597,5598..5666))
                     /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     5429..5448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5667..5771)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5797..6252)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5934..5955)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6146..6165
                     /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 ccatggtgag
     1201 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1261 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1321 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1381 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1441 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1501 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1561 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1621 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1681 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1741 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1801 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1861 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc acccagcttt
     1921 cttgtacaaa gtggctgcag agatgggcag taaaggggtg acggcgggaa agatcgccag
     1981 caacgtgcag aagaagctca cccgcgcgca ggagaaggtt ctccagaagc tggggaaggc
     2041 agatgagacc aaggatgagc agtttgagca gtgcgtccag aatttcaaca agcagctgac
     2101 ggagggcacc cggctgcaga aggatctccg gacctacctg gcctccgtca aagccatgca
     2161 cgaggcttcc aagaagctga atgagtgtct gcaggaggtg tatgagcccg attggcccgg
     2221 cagggatgag gcaaacaaga tcgcagagaa caacgacctg ctgtggatgg attaccacca
     2281 gaagctggtg gaccaggcgc tgctgaccat ggacacgtac ctgggccagt tccccgacat
     2341 caagtcacgc attgccaagc gggggcgcaa gctggtggac tacgacagtg cccggcacca
     2401 ctacgagtcc cttcaaactg ccaaaaagaa ggatgaagcc aaaattgcca aggccgagga
     2461 ggagctcatc aaagcccaga aggtgtttga ggagatgaat gtggatctgc aggaggagct
     2521 gccgtccctg tggaacagcc gcgtaggttt ctacgtcaac acgttccaga gcatcgcggg
     2581 cctggaggaa aacttccaca aggagatgag caagctcaac cagaacctca atgatgtgct
     2641 ggtcggcctg gagaagcaac acgggagcaa caccttcacg gtcaaggccc agcccagaaa
     2701 gaaaagtaaa ctgttttcgc ggctgcgcag aaagaagaac agtgacaacg cgcctgcaaa
     2761 agggaacaag agcccttcgc ctccagatgg ctcccctgcc gccacccccg agatcagagt
     2821 caaccacgag ccagagccgg ccggcggggc cacgcccggg gccaccctcc ccaagtcccc
     2881 atctcagcca gcagaggcct cggaggtggc gggtgggacc caacctgcgg ctggagccca
     2941 ggagccaggg gagacggcgg caagtgaagc agcctccagc tctcttcctg ctgtcgtggt
     3001 ggagaccttc ccagcaactg tgaatggcac cgtggagggc ggcagtgggg ccgggcgctt
     3061 ggacctgccc ccaggtttca tgttcaaggt acaggcccag cacgactaca cggccactga
     3121 cacagacgag ctgcagctca aggctggtga tgtggtgctg gtgatcccct tccagaaccc
     3181 tgaagagcag gatgaaggct ggctcatggg cgtgaaggag agcgactgga accagcacaa
     3241 ggagctggag aagtgccgtg gcgtcttccc cgagaacttc actgagaggg tcccatgaca
     3301 actttattat acatagttgc actcgagcct ctagaactat agtgagtcgt attacgtaga
     3361 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3421 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3481 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3541 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     3601 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     3661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     3721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5761 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     5821 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5881 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     5941 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     6001 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     6061 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6121 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6181 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6241 tacagggcgc gtc
//
