LOCUS       sequence-176844-        9025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  research of alternative isoforms of the protein dysferlin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9025)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 9025)
  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"
COMMENT     Sequence Label: pUC57 DYSF, alt exon 1, 5a, 17, 40a
FEATURES             Location/Qualifiers
     source          1..9025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..29
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13..29
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             821..832
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6483..6499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6483..6499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6496..6518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6507..6523
                     /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(6531..6537,6538..6555,6556..6561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6576..6597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6714..6731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6885..7473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6965..6984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7644..8459)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7644..8459)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     8367..8386
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8460..8564)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8632..8650
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8688..8710)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8810..8829
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(9023..9025,1..20)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agtcacgacg ttgtaaaacg acggccagtg aattgacgcg tattgggata tgctgagggt
       61 cttcatcctc tatgccgaga acgtccacac acccgacacc gacatcagcg atgcctactg
      121 ctccgcggtg tttgcagggg tgaagaagag aaccaaagtc atcaagaaca gcgtgaaccc
      181 tgtatggaat gagggatttg aatgggacct caagggcatc cccctggacc agggctctga
      241 gcttcatgtg gtggtcaaag accatgagac gatggggagg aacaggttcc tgggggaagc
      301 caaggtccca ctccgagagg tcctcgccac ccctagtctg tccgccagct tcaatgcccc
      361 cctgctggac accaagaagc agcccacagg ggcctcgctg gtcctgcagg tgtcctacac
      421 accgctgcct ggagctgtgc ccctgttccc gccccctact cctctggagc cctccccgac
      481 tctgcctgac ctggatgtag tggcaggcgg gggacagagc cgggccgaga cttggtccct
      541 gctcagtgac agcaccatgg acacgagata ctctggaaag aagtggccgg cccccacgga
      601 cacaggagga gaggaagaca cagaggacca gggactcact ggagatgagg cggagccatt
      661 cctggatcaa agcggaggcc cgggggctcc caccacccca aggaaactac cttcacgtcc
      721 tccgccccac taccccggga tcaaaagaaa gcgaagtgcg cctacatcta gaaagctgct
      781 gtcagacaaa ccgcaggatt tccagatcag ggtccaggtg atcgaggggc gccagctgcc
      841 gggggtgaac atcaagcctg tggtcaaggt taccgctgca gggcagacca agcggacgcg
      901 gatccacaag ggaaacagcc cactcttcaa tgagactctt ttcttcaact tgtttgactc
      961 tcctggggag ctgtttgatg agcccatctt tatcacggtg gtagactctc gttctctcag
     1021 gacagatgct ctcctcgggg agttccggat ggacgtgggc accatttaca gagagccccg
     1081 gcacgcctat ctcaggaagt ggctgctgct ctcagaccct gatgacttct ctgctggggc
     1141 cagaggctac ctgaaaacaa gcctttgtgt gctggggcct ggggacgaag cgcctctgga
     1201 gagaaaagac ccctctgaag acaaggagga cattgaaagc aacctgctcc ggcccacagg
     1261 cgtagccctg cgaggagccc acttctgcct gaaggtcttc cgggccgagg acttgccgca
     1321 gatggacgat gccgtgatgg acaacgtgaa acagatcttt ggcttcgaga gtaacaagaa
     1381 gaacttggtg gacccctttg tggaggtcag ctttgcgggg aaaatgctgt gcagcaagat
     1441 cttggagaag acggccaacc ctcagtggaa ccagaacatc acactgcctg ccatgtttcc
     1501 ctccatgtgc gaaaaaatga ggattcgtat catagactgg gaccgcctga ctcacaatga
     1561 catcgtggct accacctacc tgagtatgtc gaaaatctct gcccctggag gagaaataga
     1621 agaggagcct gcaggtgctg tcaagccttc gaaagcctca gacttggatg actacctggg
     1681 cttcctcccc acttttgggc cctgctacat caacctctat ggcagtccca gagagttcac
     1741 aggcttccca gacccctaca cagagctcaa cacaggcaag ggggaaggtg tggcttatcg
     1801 tggccggctt ctgctctccc tggagaccaa gctggtggag cacagtgaac agaaggtgga
     1861 ggaccttcct gcggatgaca tcctccgggt ggagaagtac cttaggaggc gcaagtactc
     1921 cctgtttgcg gccttctact cagccaccat gctgcaggat gtggatgacg ccatccagtt
     1981 tgaggtcagc atcgggaact acgggaacaa gttcgacatg acctgcctgc cgctggcctc
     2041 caccactcag tacagccgtg cagtctttga cgggtgccac tactactacc taccctgggg
     2101 taacgtgaaa cccgtggtgg tgctgtcatc ctactgggag gacatcagcc atagaatcga
     2161 gactcagaac cagctgcttg ggattgctga ccggctggaa gctggcctgg agcaggtcca
     2221 cctggccctg aaggcgcagt gctccacgga ggacgtggac tcgctggtgg ctcagctgac
     2281 ggatgagctc atcgcaggct gcagccagcc tctgggtgac atccatgaga caccctctgc
     2341 cacccacctg gaccagtacc tgtaccagct gcgcacccat cacctgagcc aaatcactga
     2401 ggctgccctg gccctgaagc tcggccacag tgagctccct gcagctctgg agcaggcgga
     2461 ggactggctc ctgcgtctgc gtgccctggc agaggagccc cagaacagcc tgccggacat
     2521 cgtcatctgg atgctgcagg gagacaagcg tgtggcatac cagcgggtgc ctgcccacca
     2581 agtcctcttc tcccggcggg gtgccaacta ctgtggcaag aattgtggga agctacagac
     2641 aatctttctg aaatatccga tggagaaggt gcctggcgcc cggatgccag tgcagatacg
     2701 ggtcaagctg tggtttgggc tctctgtgga tgagaaggag ttcaaccagt ttgctgaggg
     2761 gaagctgtct gtctttgctg aaacctatga gaacgagact aagttggccc ttgttgggaa
     2821 ctggggcaca acgggcctca cctaccccaa gttttctgac gtcacgggca agatcaagct
     2881 acccaaggac agcttccgcc cctcggccgg ctggacctgg gctggagatt ggttcgtgtg
     2941 cccggagaag actctgctcc atgacatgga cgccggtcac ctgagcttcg tggaagaggt
     3001 gtttgagaac cagacccggc ttcccggagg ccagtggatc tacatgagtg acaactacac
     3061 cgatgtgaac ggggagaagg tgcttcccaa ggatgacatt gagtgcccac tgggctggaa
     3121 gtgggaagat gaggaatggt ccacagacct caaccgggct gtcgatgagc aaggctggga
     3181 gtatagcatc accatccccc cggagcggaa gccgaagcac tgggtccctg ctgagaagat
     3241 gtactacaca caccgacggc ggcgctgggt gcgcctgcgc aggagggatc tcagccaaat
     3301 ggaagcactg aaaaggcaca ggcaggcgga ggcggagggc gagggctggg agtacgcctc
     3361 tctttttggc tggaagttcc acctcgagta ccgcaagaca gatgccttcc gccgccgccg
     3421 ctggcgccgt cgcatggagc cactggagaa gacggggcct gcagctgtgt ttgcccttga
     3481 gggggccctg ggcggcgtga tggatgacaa gagtgaagat tccatgtccg tctccacctt
     3541 gagcttcggt gtgaacagac ccacgatttc ctgcatattc gactatggga accgctacca
     3601 tctacgctgc tacatgtacc aggcccggga cctggctgcg atggacaagg actctttttc
     3661 tgatccctat gccatcgtct ccttcctgca ccagagccag aagacggtgg tggtgaagaa
     3721 cacccttaac cccacctggg accagacgct catcttctac gagatcgaga tctttggcga
     3781 gccggccaca gttgctgagc aaccgcccag cattgtggtg gagctgtacg accatgacac
     3841 ttatggtgca gacgagttta tgggtcgctg catctgtcaa ccgagtctgg aacggatgcc
     3901 acggctggcc tggttcccac tgacgagggg cagccagccg tcgggggagc tgctggcctc
     3961 ttttgagctc atccagagag agaagccggc catccaccat attcctggtt ttgaggtgca
     4021 ggagacatca aggatcctgg atgagtctga ggacacagac ctgccctacc caccacccca
     4081 gagggaggcc aacatctaca tggttcctca gaacatcaag ccagcgctcc agcgtaccgc
     4141 catcgagatc ctggcatggg gcctgcggaa catgaagagt taccagctgg ccaacatctc
     4201 ctcccccagc ctcgtggtag agtgtggggg ccagacggtg cagtcctgtg tcatcaggaa
     4261 cctccggaag aaccccaact ttgacatctg caccctcttc atggaagtga tgctgcccag
     4321 ggaggagctc tactgccccc ccatcaccgt caaggtcatc gataaccgcc agtttggccg
     4381 ccggcctgtg gtgggccagt gtaccatccg ctccctggag agcttcctgt gtgaccccta
     4441 ctcggcggag agtccatccc cacagggtgg cccagacgat gtgagcctac tcagtcctgg
     4501 ggaagacgtg ctcatcgaca ttgatgacaa ggagcccctc atccccatcc agcttgcaga
     4561 cggtctgtcg agcttggccc ccactaacac ggcttctcct ccatccagtc ctcatgagga
     4621 agagttcatc gattggtgga gcaaattctt tgcctccata ggggagaggg aaaagtgcgg
     4681 ctcctacctg gagaaggatt ttgacaccct gaaggtctat gacacacagc tggagaatgt
     4741 ggaggccttt gagggcctgt ctgacttttg taacaccttc aagctgtacc ggggcaagac
     4801 gcaggaggag acagaagatc catctgtgat tggtgaattt aagggcctct tcaaaattta
     4861 tcccctccca gaagacccag ccatccccat gcccccaaga cagttccacc agctggccgc
     4921 ccagggaccc caggagtgct tggtccgtat ctacattgtc cgagcatttg gcctgcagcc
     4981 caaggacccc aatggaaagt gtgatcctta catcaagatc tccataggga agaaatcagt
     5041 gagtgaccag gataactaca tcccctgcac gctggagccc gtatttggaa agatgttcga
     5101 gctgacctgc actctgcctc tggagaagga cctaaagatc actctctatg actatgacct
     5161 cctctccaag gacgaaaaga tcggtgagac ggtcgtcgac ctggagaaca ggctgctgtc
     5221 caagtttggg gctcgctgtg gactcccaca gacctactgt gtctctggac cgaaccagtg
     5281 gcgggaccag ctccgcccct cccagctcct ccacctcttc tgccagcagc atagagtcaa
     5341 ggcacctgtg taccggacag accgtgtaat gtttcaggat aaagaatatt ccattgaaga
     5401 gatagaggct ggcaggatcc caaacccaca cctgggccca gtggaggagc gtctggctct
     5461 gcatgtgctt cagcagcagg gcctggtccc ggagcacgtg gagtcacggc ccctctacag
     5521 ccccctgcag ccagacatcg agcaggggaa gctgcagatg tgggtcgacc tatttccgaa
     5581 ggccctgggg cggcctggac ctcccttcaa catcacccca cggagagcca gaaggttttt
     5641 cctgcgttgt attatctgga ataccagaga tgtgatcctg gatgacctga gcctcacggg
     5701 ggagaagatg agcgacattt atgtgaaagg ttggatgatt ggctttgaag aacacaagca
     5761 aaagacagac gtgcattatc gttccctggg aggtgaaggc aacttcaact ggaggttcat
     5821 tttccccttc gactacctgc cagctgagca agtctgtacc attgccaaga aggatgcctt
     5881 ctggaggctg gacaagactg agagcaaaat cccagcacga gtggtgttcc agatctggga
     5941 caatgacaag ttctcctttg atgattttct gggctccctg cagctcgatc tcaaccgcat
     6001 gcccaagcca gccaagacag ccaagaagtg ctccttggac cagctggatg atgctttcca
     6061 cccagaatgg tttgtgtccc tttttgagca gaaaacagtg aagggctggt ggccctgtgt
     6121 agcagaagag ggtgagaaga aaatactggc gggcaagctg gaaatgacct tggagattgt
     6181 agcagagagt gagcatgagg agcggcctgc tggccagggc cgggatgagc ccaacatgaa
     6241 ccctaagctt gaggacccaa ggcgccccga cacctccttc ctgtggttta cctccccata
     6301 caagaccatg aagttcatcc tgtggcggcg tttccggtgg gccatcatcc tcttcatcat
     6361 cctcttcatc ctgctgctgt tcctggccat cttcatctac gccttcccga actatgctgc
     6421 catgaagctg gtgaagccct tcagctgaat cccaatggcg cgccgagctt ggctcgagca
     6481 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6541 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6601 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7621 tatgagtaaa cttggtctga cagttagaaa aactcatcga gcatcaaatg aaactgcaat
     7681 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     7741 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     7801 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     7861 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     7921 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     7981 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     8041 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     8101 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccagggatc
     8161 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     8221 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     8281 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     8341 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     8401 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     8461 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8521 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8581 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     8641 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     8701 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     8761 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     8821 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     8881 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     8941 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     9001 agttgggtaa cgccagggtt ttccc
//
