LOCUS       sequence-174248-        8978 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 8978)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8978)
  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 1a, 5a, 40a
FEATURES             Location/Qualifiers
     source          1..8978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..35
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     27..44
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     28..44
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             831..842
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6451..6467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6451..6467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6464..6486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6475..6491
                     /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(6499..6505,6506..6523,6524..6529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6544..6565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6682..6699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6853..7441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6933..6952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7612..8427)
                     /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(7612..8427)
                     /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     8335..8354
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8428..8532)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8600..8618
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8656..8678)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8778..8797
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg acgcgtatgc
       61 tgtgctgcct gctggtgagg gccagcaacc tccccagtgc gaagaaggac cggcgcagcg
      121 accctgtcgc aagcctgact ttccgagggg tgaagaagag aaccaaagtc atcaagaaca
      181 gcgtgaaccc tgtatggaat gagggatttg aatgggacct caagggcatc cccctggacc
      241 agggctctga gcttcatgtg gtggtcaaag accatgagac gatggggagg aacaggttcc
      301 tgggggaagc caaggtccca ctccgagagg tcctcgccac ccctagtctg tccgccagct
      361 tcaatgcccc cctgctggac accaagaagc agcccacagg ggcctcgctg gtcctgcagg
      421 tgtcctacac accgctgcct ggagctgtgc ccctgttccc gccccctact cctctggagc
      481 cctccccgac tctgcctgac ctggatgtag tggcaggcgg gggacagagc cgggccgaga
      541 cttggtccct gctcagtgac agcaccatgg acacgagata ctctggaaag aagtggccgg
      601 cccccacgga cacaggagga gaggaagaca cagaggacca gggactcact ggagatgagg
      661 cggagccatt cctggatcaa agcggaggcc cgggggctcc caccacccca aggaaactac
      721 cttcacgtcc tccgccccac taccccggga tcaaaagaaa gcgaagtgcg cctacatcta
      781 gaaagctgct gtcagacaaa ccgcaggatt tccagatcag ggtccaggtg atcgaggggc
      841 gccagctgcc gggggtgaac atcaagcctg tggtcaaggt taccgctgca gggcagacca
      901 agcggacgcg gatccacaag ggaaacagcc cactcttcaa tgagactctt ttcttcaact
      961 tgtttgactc tcctggggag ctgtttgatg agcccatctt tatcacggtg gtagactctc
     1021 gttctctcag gacagatgct ctcctcgggg agttccggat ggacgtgggc accatttaca
     1081 gagagccccg gcacgcctat ctcaggaagt ggctgctgct ctcagaccct gatgacttct
     1141 ctgctggggc cagaggctac ctgaaaacaa gcctttgtgt gctggggcct ggggacgaag
     1201 cgcctctgga gagaaaagac ccctctgaag acaaggagga cattgaaagc aacctgctcc
     1261 ggcccacagg cgtagccctg cgaggagccc acttctgcct gaaggtcttc cgggccgagg
     1321 acttgccgca gatggacgat gccgtgatgg acaacgtgaa acagatcttt ggcttcgaga
     1381 gtaacaagaa gaacttggtg gacccctttg tggaggtcag ctttgcgggg aaaatgctgt
     1441 gcagcaagat cttggagaag acggccaacc ctcagtggaa ccagaacatc acactgcctg
     1501 ccatgtttcc ctccatgtgc gaaaaaatga ggattcgtat catagactgg gaccgcctga
     1561 ctcacaatga catcgtggct accacctacc tgagtatgtc gaaaatctct gcccctggag
     1621 gagaaataga agtggatgac tacctgggct tcctccccac ttttgggccc tgctacatca
     1681 acctctatgg cagtcccaga gagttcacag gcttcccaga cccctacaca gagctcaaca
     1741 caggcaaggg ggaaggtgtg gcttatcgtg gccggcttct gctctccctg gagaccaagc
     1801 tggtggagca cagtgaacag aaggtggagg accttcctgc ggatgacatc ctccgggtgg
     1861 agaagtacct taggaggcgc aagtactccc tgtttgcggc cttctactca gccaccatgc
     1921 tgcaggatgt ggatgacgcc atccagtttg aggtcagcat cgggaactac gggaacaagt
     1981 tcgacatgac ctgcctgccg ctggcctcca ccactcagta cagccgtgca gtctttgacg
     2041 ggtgccacta ctactaccta ccctggggta acgtgaaacc cgtggtggtg ctgtcatcct
     2101 actgggagga catcagccat agaatcgaga ctcagaacca gctgcttggg attgctgacc
     2161 ggctggaagc tggcctggag caggtccacc tggccctgaa ggcgcagtgc tccacggagg
     2221 acgtggactc gctggtggct cagctgacgg atgagctcat cgcaggctgc agccagcctc
     2281 tgggtgacat ccatgagaca ccctctgcca cccacctgga ccagtacctg taccagctgc
     2341 gcacccatca cctgagccaa atcactgagg ctgccctggc cctgaagctc ggccacagtg
     2401 agctccctgc agctctggag caggcggagg actggctcct gcgtctgcgt gccctggcag
     2461 aggagcccca gaacagcctg ccggacatcg tcatctggat gctgcaggga gacaagcgtg
     2521 tggcatacca gcgggtgcct gcccaccaag tcctcttctc ccggcggggt gccaactact
     2581 gtggcaagaa ttgtgggaag ctacagacaa tctttctgaa atatccgatg gagaaggtgc
     2641 ctggcgcccg gatgccagtg cagatacggg tcaagctgtg gtttgggctc tctgtggatg
     2701 agaaggagtt caaccagttt gctgagggga agctgtctgt ctttgctgaa acctatgaga
     2761 acgagactaa gttggccctt gttgggaact ggggcacaac gggcctcacc taccccaagt
     2821 tttctgacgt cacgggcaag atcaagctac ccaaggacag cttccgcccc tcggccggct
     2881 ggacctgggc tggagattgg ttcgtgtgcc cggagaagac tctgctccat gacatggacg
     2941 ccggtcacct gagcttcgtg gaagaggtgt ttgagaacca gacccggctt cccggaggcc
     3001 agtggatcta catgagtgac aactacaccg atgtgaacgg ggagaaggtg cttcccaagg
     3061 atgacattga gtgcccactg ggctggaagt gggaagatga ggaatggtcc acagacctca
     3121 accgggctgt cgatgagcaa ggctgggagt atagcatcac catccccccg gagcggaagc
     3181 cgaagcactg ggtccctgct gagaagatgt actacacaca ccgacggcgg cgctgggtgc
     3241 gcctgcgcag gagggatctc agccaaatgg aagcactgaa aaggcacagg caggcggagg
     3301 cggagggcga gggctgggag tacgcctctc tttttggctg gaagttccac ctcgagtacc
     3361 gcaagacaga tgccttccgc cgccgccgct ggcgccgtcg catggagcca ctggagaaga
     3421 cggggcctgc agctgtgttt gcccttgagg gggccctggg cggcgtgatg gatgacaaga
     3481 gtgaagattc catgtccgtc tccaccttga gcttcggtgt gaacagaccc acgatttcct
     3541 gcatattcga ctatgggaac cgctaccatc tacgctgcta catgtaccag gcccgggacc
     3601 tggctgcgat ggacaaggac tctttttctg atccctatgc catcgtctcc ttcctgcacc
     3661 agagccagaa gacggtggtg gtgaagaaca cccttaaccc cacctgggac cagacgctca
     3721 tcttctacga gatcgagatc tttggcgagc cggccacagt tgctgagcaa ccgcccagca
     3781 ttgtggtgga gctgtacgac catgacactt atggtgcaga cgagtttatg ggtcgctgca
     3841 tctgtcaacc gagtctggaa cggatgccac ggctggcctg gttcccactg acgaggggca
     3901 gccagccgtc gggggagctg ctggcctctt ttgagctcat ccagagagag aagccggcca
     3961 tccaccatat tcctggtttt gaggtgcagg agacatcaag gatcctggat gagtctgagg
     4021 acacagacct gccctaccca ccaccccaga gggaggccaa catctacatg gttcctcaga
     4081 acatcaagcc agcgctccag cgtaccgcca tcgagatcct ggcatggggc ctgcggaaca
     4141 tgaagagtta ccagctggcc aacatctcct cccccagcct cgtggtagag tgtgggggcc
     4201 agacggtgca gtcctgtgtc atcaggaacc tccggaagaa ccccaacttt gacatctgca
     4261 ccctcttcat ggaagtgatg ctgcccaggg aggagctcta ctgccccccc atcaccgtca
     4321 aggtcatcga taaccgccag tttggccgcc ggcctgtggt gggccagtgt accatccgct
     4381 ccctggagag cttcctgtgt gacccctact cggcggagag tccatcccca cagggtggcc
     4441 cagacgatgt gagcctactc agtcctgggg aagacgtgct catcgacatt gatgacaagg
     4501 agcccctcat ccccatccag cttgcagacg gtctgtcgag cttggccccc actaacacgg
     4561 cttctcctcc atccagtcct catgaggaag agttcatcga ttggtggagc aaattctttg
     4621 cctccatagg ggagagggaa aagtgcggct cctacctgga gaaggatttt gacaccctga
     4681 aggtctatga cacacagctg gagaatgtgg aggcctttga gggcctgtct gacttttgta
     4741 acaccttcaa gctgtaccgg ggcaagacgc aggaggagac agaagatcca tctgtgattg
     4801 gtgaatttaa gggcctcttc aaaatttatc ccctcccaga agacccagcc atccccatgc
     4861 ccccaagaca gttccaccag ctggccgccc agggacccca ggagtgcttg gtccgtatct
     4921 acattgtccg agcatttggc ctgcagccca aggaccccaa tggaaagtgt gatccttaca
     4981 tcaagatctc catagggaag aaatcagtga gtgaccagga taactacatc ccctgcacgc
     5041 tggagcccgt atttggaaag atgttcgagc tgacctgcac tctgcctctg gagaaggacc
     5101 taaagatcac tctctatgac tatgacctcc tctccaagga cgaaaagatc ggtgagacgg
     5161 tcgtcgacct ggagaacagg ctgctgtcca agtttggggc tcgctgtgga ctcccacaga
     5221 cctactgtgt ctctggaccg aaccagtggc gggaccagct ccgcccctcc cagctcctcc
     5281 acctcttctg ccagcagcat agagtcaagg cacctgtgta ccggacagac cgtgtaatgt
     5341 ttcaggataa agaatattcc attgaagaga tagaggctgg caggatccca aacccacacc
     5401 tgggcccagt ggaggagcgt ctggctctgc atgtgcttca gcagcagggc ctggtcccgg
     5461 agcacgtgga gtcacggccc ctctacagcc ccctgcagcc agacatcgag caggggaagc
     5521 tgcagatgtg ggtcgaccta tttccgaagg ccctggggcg gcctggacct cccttcaaca
     5581 tcaccccacg gagagccaga aggtttttcc tgcgttgtat tatctggaat accagagatg
     5641 tgatcctgga tgacctgagc ctcacggggg agaagatgag cgacatttat gtgaaaggtt
     5701 ggatgattgg ctttgaagaa cacaagcaaa agacagacgt gcattatcgt tccctgggag
     5761 gtgaaggcaa cttcaactgg aggttcattt tccccttcga ctacctgcca gctgagcaag
     5821 tctgtaccat tgccaagaag gatgccttct ggaggctgga caagactgag agcaaaatcc
     5881 cagcacgagt ggtgttccag atctgggaca atgacaagtt ctcctttgat gattttctgg
     5941 gctccctgca gctcgatctc aaccgcatgc ccaagccagc caagacagcc aagaagtgct
     6001 ccttggacca gctggatgat gctttccacc cagaatggtt tgtgtccctt tttgagcaga
     6061 aaacagtgaa gggctggtgg ccctgtgtag cagaagaggg tgagaagaaa atactggcgg
     6121 gcaagctgga aatgaccttg gagattgtag cagagagtga gcatgaggag cggcctgctg
     6181 gccagggccg ggatgagccc aacatgaacc ctaagcttga ggacccaagg cgccccgaca
     6241 cctccttcct gtggtttacc tccccataca agaccatgaa gttcatcctg tggcggcgtt
     6301 tccggtgggc catcatcctc ttcatcatcc tcttcatcct gctgctgttc ctggccatct
     6361 tcatctacgc cttcccgaac tatgctgcca tgaagctggt gaagcccttc agctgaatcc
     6421 caatggcgcg ccgagcttgg ctcgagcatg gtcatagctg tttcctgtgt gaaattgtta
     6481 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6541 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6601 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6661 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6721 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6781 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7021 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7261 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7501 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7561 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttagaaaaa
     7621 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     7681 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     7741 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     7801 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     7861 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     7921 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     7981 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     8041 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     8101 tacctggaat gctgttttcc cagggatcgc agtggtgagt aaccatgcat catcaggagt
     8161 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     8221 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     8281 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     8341 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     8401 agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta
     8461 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8521 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8581 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8641 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     8701 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     8761 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     8821 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     8881 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     8941 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaa
//
