LOCUS       sequence-174216-        8983 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 8983)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8983)
  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, 40a
FEATURES             Location/Qualifiers
     source          1..8983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..40
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     32..49
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     33..49
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             841..852
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6461..6477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6461..6477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6474..6496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6485..6501
                     /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(6509..6515,6516..6533,6534..6539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6554..6575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6692..6709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6863..7451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6943..6962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7622..8437)
                     /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(7622..8437)
                     /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     8345..8364
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8438..8542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8610..8628
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8666..8688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8788..8807
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgacgcg
       61 tattgggata tgctgagggt cttcatcctc tatgccgaga acgtccacac acccgacacc
      121 gacatcagcg atgcctactg ctccgcggtg tttgcagggg tgaagaagag aaccaaagtc
      181 atcaagaaca gcgtgaaccc tgtatggaat gagggatttg aatgggacct caagggcatc
      241 cccctggacc agggctctga gcttcatgtg gtggtcaaag accatgagac gatggggagg
      301 aacaggttcc tgggggaagc caaggtccca ctccgagagg tcctcgccac ccctagtctg
      361 tccgccagct tcaatgcccc cctgctggac accaagaagc agcccacagg ggcctcgctg
      421 gtcctgcagg tgtcctacac accgctgcct ggagctgtgc ccctgttccc gccccctact
      481 cctctggagc cctccccgac tctgcctgac ctggatgtag tggcaggcgg gggacagagc
      541 cgggccgaga cttggtccct gctcagtgac agcaccatgg acacgagata ctctggaaag
      601 aagtggccgg cccccacgga cacaggagga gaggaagaca cagaggacca gggactcact
      661 ggagatgagg cggagccatt cctggatcaa agcggaggcc cgggggctcc caccacccca
      721 aggaaactac cttcacgtcc tccgccccac taccccggga tcaaaagaaa gcgaagtgcg
      781 cctacatcta gaaagctgct gtcagacaaa ccgcaggatt tccagatcag ggtccaggtg
      841 atcgaggggc gccagctgcc gggggtgaac atcaagcctg tggtcaaggt taccgctgca
      901 gggcagacca agcggacgcg gatccacaag ggaaacagcc cactcttcaa tgagactctt
      961 ttcttcaact tgtttgactc tcctggggag ctgtttgatg agcccatctt tatcacggtg
     1021 gtagactctc gttctctcag gacagatgct ctcctcgggg agttccggat ggacgtgggc
     1081 accatttaca gagagccccg gcacgcctat ctcaggaagt ggctgctgct ctcagaccct
     1141 gatgacttct ctgctggggc cagaggctac ctgaaaacaa gcctttgtgt gctggggcct
     1201 ggggacgaag cgcctctgga gagaaaagac ccctctgaag acaaggagga cattgaaagc
     1261 aacctgctcc ggcccacagg cgtagccctg cgaggagccc acttctgcct gaaggtcttc
     1321 cgggccgagg acttgccgca gatggacgat gccgtgatgg acaacgtgaa acagatcttt
     1381 ggcttcgaga gtaacaagaa gaacttggtg gacccctttg tggaggtcag ctttgcgggg
     1441 aaaatgctgt gcagcaagat cttggagaag acggccaacc ctcagtggaa ccagaacatc
     1501 acactgcctg ccatgtttcc ctccatgtgc gaaaaaatga ggattcgtat catagactgg
     1561 gaccgcctga ctcacaatga catcgtggct accacctacc tgagtatgtc gaaaatctct
     1621 gcccctggag gagaaataga agtggatgac tacctgggct tcctccccac ttttgggccc
     1681 tgctacatca acctctatgg cagtcccaga gagttcacag gcttcccaga cccctacaca
     1741 gagctcaaca caggcaaggg ggaaggtgtg gcttatcgtg gccggcttct gctctccctg
     1801 gagaccaagc tggtggagca cagtgaacag aaggtggagg accttcctgc ggatgacatc
     1861 ctccgggtgg agaagtacct taggaggcgc aagtactccc tgtttgcggc cttctactca
     1921 gccaccatgc tgcaggatgt ggatgacgcc atccagtttg aggtcagcat cgggaactac
     1981 gggaacaagt tcgacatgac ctgcctgccg ctggcctcca ccactcagta cagccgtgca
     2041 gtctttgacg ggtgccacta ctactaccta ccctggggta acgtgaaacc cgtggtggtg
     2101 ctgtcatcct actgggagga catcagccat agaatcgaga ctcagaacca gctgcttggg
     2161 attgctgacc ggctggaagc tggcctggag caggtccacc tggccctgaa ggcgcagtgc
     2221 tccacggagg acgtggactc gctggtggct cagctgacgg atgagctcat cgcaggctgc
     2281 agccagcctc tgggtgacat ccatgagaca ccctctgcca cccacctgga ccagtacctg
     2341 taccagctgc gcacccatca cctgagccaa atcactgagg ctgccctggc cctgaagctc
     2401 ggccacagtg agctccctgc agctctggag caggcggagg actggctcct gcgtctgcgt
     2461 gccctggcag aggagcccca gaacagcctg ccggacatcg tcatctggat gctgcaggga
     2521 gacaagcgtg tggcatacca gcgggtgcct gcccaccaag tcctcttctc ccggcggggt
     2581 gccaactact gtggcaagaa ttgtgggaag ctacagacaa tctttctgaa atatccgatg
     2641 gagaaggtgc ctggcgcccg gatgccagtg cagatacggg tcaagctgtg gtttgggctc
     2701 tctgtggatg agaaggagtt caaccagttt gctgagggga agctgtctgt ctttgctgaa
     2761 acctatgaga acgagactaa gttggccctt gttgggaact ggggcacaac gggcctcacc
     2821 taccccaagt tttctgacgt cacgggcaag atcaagctac ccaaggacag cttccgcccc
     2881 tcggccggct ggacctgggc tggagattgg ttcgtgtgcc cggagaagac tctgctccat
     2941 gacatggacg ccggtcacct gagcttcgtg gaagaggtgt ttgagaacca gacccggctt
     3001 cccggaggcc agtggatcta catgagtgac aactacaccg atgtgaacgg ggagaaggtg
     3061 cttcccaagg atgacattga gtgcccactg ggctggaagt gggaagatga ggaatggtcc
     3121 acagacctca accgggctgt cgatgagcaa ggctgggagt atagcatcac catccccccg
     3181 gagcggaagc cgaagcactg ggtccctgct gagaagatgt actacacaca ccgacggcgg
     3241 cgctgggtgc gcctgcgcag gagggatctc agccaaatgg aagcactgaa aaggcacagg
     3301 caggcggagg cggagggcga gggctgggag tacgcctctc tttttggctg gaagttccac
     3361 ctcgagtacc gcaagacaga tgccttccgc cgccgccgct ggcgccgtcg catggagcca
     3421 ctggagaaga cggggcctgc agctgtgttt gcccttgagg gggccctggg cggcgtgatg
     3481 gatgacaaga gtgaagattc catgtccgtc tccaccttga gcttcggtgt gaacagaccc
     3541 acgatttcct gcatattcga ctatgggaac cgctaccatc tacgctgcta catgtaccag
     3601 gcccgggacc tggctgcgat ggacaaggac tctttttctg atccctatgc catcgtctcc
     3661 ttcctgcacc agagccagaa gacggtggtg gtgaagaaca cccttaaccc cacctgggac
     3721 cagacgctca tcttctacga gatcgagatc tttggcgagc cggccacagt tgctgagcaa
     3781 ccgcccagca ttgtggtgga gctgtacgac catgacactt atggtgcaga cgagtttatg
     3841 ggtcgctgca tctgtcaacc gagtctggaa cggatgccac ggctggcctg gttcccactg
     3901 acgaggggca gccagccgtc gggggagctg ctggcctctt ttgagctcat ccagagagag
     3961 aagccggcca tccaccatat tcctggtttt gaggtgcagg agacatcaag gatcctggat
     4021 gagtctgagg acacagacct gccctaccca ccaccccaga gggaggccaa catctacatg
     4081 gttcctcaga acatcaagcc agcgctccag cgtaccgcca tcgagatcct ggcatggggc
     4141 ctgcggaaca tgaagagtta ccagctggcc aacatctcct cccccagcct cgtggtagag
     4201 tgtgggggcc agacggtgca gtcctgtgtc atcaggaacc tccggaagaa ccccaacttt
     4261 gacatctgca ccctcttcat ggaagtgatg ctgcccaggg aggagctcta ctgccccccc
     4321 atcaccgtca aggtcatcga taaccgccag tttggccgcc ggcctgtggt gggccagtgt
     4381 accatccgct ccctggagag cttcctgtgt gacccctact cggcggagag tccatcccca
     4441 cagggtggcc cagacgatgt gagcctactc agtcctgggg aagacgtgct catcgacatt
     4501 gatgacaagg agcccctcat ccccatccag cttgcagacg gtctgtcgag cttggccccc
     4561 actaacacgg cttctcctcc atccagtcct catgaggaag agttcatcga ttggtggagc
     4621 aaattctttg cctccatagg ggagagggaa aagtgcggct cctacctgga gaaggatttt
     4681 gacaccctga aggtctatga cacacagctg gagaatgtgg aggcctttga gggcctgtct
     4741 gacttttgta acaccttcaa gctgtaccgg ggcaagacgc aggaggagac agaagatcca
     4801 tctgtgattg gtgaatttaa gggcctcttc aaaatttatc ccctcccaga agacccagcc
     4861 atccccatgc ccccaagaca gttccaccag ctggccgccc agggacccca ggagtgcttg
     4921 gtccgtatct acattgtccg agcatttggc ctgcagccca aggaccccaa tggaaagtgt
     4981 gatccttaca tcaagatctc catagggaag aaatcagtga gtgaccagga taactacatc
     5041 ccctgcacgc tggagcccgt atttggaaag atgttcgagc tgacctgcac tctgcctctg
     5101 gagaaggacc taaagatcac tctctatgac tatgacctcc tctccaagga cgaaaagatc
     5161 ggtgagacgg tcgtcgacct ggagaacagg ctgctgtcca agtttggggc tcgctgtgga
     5221 ctcccacaga cctactgtgt ctctggaccg aaccagtggc gggaccagct ccgcccctcc
     5281 cagctcctcc acctcttctg ccagcagcat agagtcaagg cacctgtgta ccggacagac
     5341 cgtgtaatgt ttcaggataa agaatattcc attgaagaga tagaggctgg caggatccca
     5401 aacccacacc tgggcccagt ggaggagcgt ctggctctgc atgtgcttca gcagcagggc
     5461 ctggtcccgg agcacgtgga gtcacggccc ctctacagcc ccctgcagcc agacatcgag
     5521 caggggaagc tgcagatgtg ggtcgaccta tttccgaagg ccctggggcg gcctggacct
     5581 cccttcaaca tcaccccacg gagagccaga aggtttttcc tgcgttgtat tatctggaat
     5641 accagagatg tgatcctgga tgacctgagc ctcacggggg agaagatgag cgacatttat
     5701 gtgaaaggtt ggatgattgg ctttgaagaa cacaagcaaa agacagacgt gcattatcgt
     5761 tccctgggag gtgaaggcaa cttcaactgg aggttcattt tccccttcga ctacctgcca
     5821 gctgagcaag tctgtaccat tgccaagaag gatgccttct ggaggctgga caagactgag
     5881 agcaaaatcc cagcacgagt ggtgttccag atctgggaca atgacaagtt ctcctttgat
     5941 gattttctgg gctccctgca gctcgatctc aaccgcatgc ccaagccagc caagacagcc
     6001 aagaagtgct ccttggacca gctggatgat gctttccacc cagaatggtt tgtgtccctt
     6061 tttgagcaga aaacagtgaa gggctggtgg ccctgtgtag cagaagaggg tgagaagaaa
     6121 atactggcgg gcaagctgga aatgaccttg gagattgtag cagagagtga gcatgaggag
     6181 cggcctgctg gccagggccg ggatgagccc aacatgaacc ctaagcttga ggacccaagg
     6241 cgccccgaca cctccttcct gtggtttacc tccccataca agaccatgaa gttcatcctg
     6301 tggcggcgtt tccggtgggc catcatcctc ttcatcatcc tcttcatcct gctgctgttc
     6361 ctggccatct tcatctacgc cttcccgaac tatgctgcca tgaagctggt gaagcccttc
     6421 agctgaatcc caatggcgcg ccgagcttgg ctcgagcatg gtcatagctg tttcctgtgt
     6481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7261 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7621 gttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     7681 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     7741 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     7801 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     7861 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     7921 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     7981 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     8041 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     8101 attcttctaa tacctggaat gctgttttcc cagggatcgc agtggtgagt aaccatgcat
     8161 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     8221 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     8281 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     8341 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     8401 gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt
     8461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8581 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     8641 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     8701 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     8761 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     8821 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     8881 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     8941 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttg
//
