LOCUS       sequence-174238-        8861 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 8861)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8861)
  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, 17
FEATURES             Location/Qualifiers
     source          1..8861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..29
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     21..38
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     22..38
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             729..740
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6328..6344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6328..6344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6341..6363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6352..6368
                     /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(6376..6382,6383..6400,6401..6406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6421..6442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6559..6576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6730..7318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6810..6829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7489..8304)
                     /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(7489..8304)
                     /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     8212..8231
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8305..8409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8477..8495
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8533..8555)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8655..8674
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgacgcgt atgctgaggg
       61 tcttcatcct ctatgccgag aacgtccaca cacccgacac cgacatcagc gatgcctact
      121 gctccgcggt gtttgcaggg gtgaagaaga gaaccaaagt catcaagaac agcgtgaacc
      181 ctgtatggaa tgagggattt gaatgggacc tcaagggcat ccccctggac cagggctctg
      241 agcttcatgt ggtggtcaaa gaccatgaga cgatggggag gaacaggttc ctgggggaag
      301 ccaaggtccc actccgagag gtcctcgcca cccctagtct gtccgccagc ttcaatgccc
      361 ccctgctgga caccaagaag cagcccacag gggcctcgct ggtcctgcag gtgtcctaca
      421 caccgctgcc tggagctgtg cccctgttcc cgccccctac tcctctggag ccctccccga
      481 ctctgcctga cctggatgta gtggcagaca caggaggaga ggaagacaca gaggaccagg
      541 gactcactgg agatgaggcg gagccattcc tggatcaaag cggaggcccg ggggctccca
      601 ccaccccaag gaaactacct tcacgtcctc cgccccacta ccccgggatc aaaagaaagc
      661 gaagtgcgcc tacatctaga aagctgctgt cagacaaacc gcaggatttc cagatcaggg
      721 tccaggtgat cgaggggcgc cagctgccgg gggtgaacat caagcctgtg gtcaaggtta
      781 ccgctgcagg gcagaccaag cggacgcgga tccacaaggg aaacagccca ctcttcaatg
      841 agactctttt cttcaacttg tttgactctc ctggggagct gtttgatgag cccatcttta
      901 tcacggtggt agactctcgt tctctcagga cagatgctct cctcggggag ttccggatgg
      961 acgtgggcac catttacaga gagccccggc acgcctatct caggaagtgg ctgctgctct
     1021 cagaccctga tgacttctct gctggggcca gaggctacct gaaaacaagc ctttgtgtgc
     1081 tggggcctgg ggacgaagcg cctctggaga gaaaagaccc ctctgaagac aaggaggaca
     1141 ttgaaagcaa cctgctccgg cccacaggcg tagccctgcg aggagcccac ttctgcctga
     1201 aggtcttccg ggccgaggac ttgccgcaga tggacgatgc cgtgatggac aacgtgaaac
     1261 agatctttgg cttcgagagt aacaagaaga acttggtgga cccctttgtg gaggtcagct
     1321 ttgcggggaa aatgctgtgc agcaagatct tggagaagac ggccaaccct cagtggaacc
     1381 agaacatcac actgcctgcc atgtttccct ccatgtgcga aaaaatgagg attcgtatca
     1441 tagactggga ccgcctgact cacaatgaca tcgtggctac cacctacctg agtatgtcga
     1501 aaatctctgc ccctggagga gaaatagaag aggagcctgc aggtgctgtc aagccttcga
     1561 aagcctcaga cttggatgac tacctgggct tcctccccac ttttgggccc tgctacatca
     1621 acctctatgg cagtcccaga gagttcacag gcttcccaga cccctacaca gagctcaaca
     1681 caggcaaggg ggaaggtgtg gcttatcgtg gccggcttct gctctccctg gagaccaagc
     1741 tggtggagca cagtgaacag aaggtggagg accttcctgc ggatgacatc ctccgggtgg
     1801 agaagtacct taggaggcgc aagtactccc tgtttgcggc cttctactca gccaccatgc
     1861 tgcaggatgt ggatgacgcc atccagtttg aggtcagcat cgggaactac gggaacaagt
     1921 tcgacatgac ctgcctgccg ctggcctcca ccactcagta cagccgtgca gtctttgacg
     1981 ggtgccacta ctactaccta ccctggggta acgtgaaacc cgtggtggtg ctgtcatcct
     2041 actgggagga catcagccat agaatcgaga ctcagaacca gctgcttggg attgctgacc
     2101 ggctggaagc tggcctggag caggtccacc tggccctgaa ggcgcagtgc tccacggagg
     2161 acgtggactc gctggtggct cagctgacgg atgagctcat cgcaggctgc agccagcctc
     2221 tgggtgacat ccatgagaca ccctctgcca cccacctgga ccagtacctg taccagctgc
     2281 gcacccatca cctgagccaa atcactgagg ctgccctggc cctgaagctc ggccacagtg
     2341 agctccctgc agctctggag caggcggagg actggctcct gcgtctgcgt gccctggcag
     2401 aggagcccca gaacagcctg ccggacatcg tcatctggat gctgcaggga gacaagcgtg
     2461 tggcatacca gcgggtgcct gcccaccaag tcctcttctc ccggcggggt gccaactact
     2521 gtggcaagaa ttgtgggaag ctacagacaa tctttctgaa atatccgatg gagaaggtgc
     2581 ctggcgcccg gatgccagtg cagatacggg tcaagctgtg gtttgggctc tctgtggatg
     2641 agaaggagtt caaccagttt gctgagggga agctgtctgt ctttgctgaa acctatgaga
     2701 acgagactaa gttggccctt gttgggaact ggggcacaac gggcctcacc taccccaagt
     2761 tttctgacgt cacgggcaag atcaagctac ccaaggacag cttccgcccc tcggccggct
     2821 ggacctgggc tggagattgg ttcgtgtgcc cggagaagac tctgctccat gacatggacg
     2881 ccggtcacct gagcttcgtg gaagaggtgt ttgagaacca gacccggctt cccggaggcc
     2941 agtggatcta catgagtgac aactacaccg atgtgaacgg ggagaaggtg cttcccaagg
     3001 atgacattga gtgcccactg ggctggaagt gggaagatga ggaatggtcc acagacctca
     3061 accgggctgt cgatgagcaa ggctgggagt atagcatcac catccccccg gagcggaagc
     3121 cgaagcactg ggtccctgct gagaagatgt actacacaca ccgacggcgg cgctgggtgc
     3181 gcctgcgcag gagggatctc agccaaatgg aagcactgaa aaggcacagg caggcggagg
     3241 cggagggcga gggctgggag tacgcctctc tttttggctg gaagttccac ctcgagtacc
     3301 gcaagacaga tgccttccgc cgccgccgct ggcgccgtcg catggagcca ctggagaaga
     3361 cggggcctgc agctgtgttt gcccttgagg gggccctggg cggcgtgatg gatgacaaga
     3421 gtgaagattc catgtccgtc tccaccttga gcttcggtgt gaacagaccc acgatttcct
     3481 gcatattcga ctatgggaac cgctaccatc tacgctgcta catgtaccag gcccgggacc
     3541 tggctgcgat ggacaaggac tctttttctg atccctatgc catcgtctcc ttcctgcacc
     3601 agagccagaa gacggtggtg gtgaagaaca cccttaaccc cacctgggac cagacgctca
     3661 tcttctacga gatcgagatc tttggcgagc cggccacagt tgctgagcaa ccgcccagca
     3721 ttgtggtgga gctgtacgac catgacactt atggtgcaga cgagtttatg ggtcgctgca
     3781 tctgtcaacc gagtctggaa cggatgccac ggctggcctg gttcccactg acgaggggca
     3841 gccagccgtc gggggagctg ctggcctctt ttgagctcat ccagagagag aagccggcca
     3901 tccaccatat tcctggtttt gaggtgcagg agacatcaag gatcctggat gagtctgagg
     3961 acacagacct gccctaccca ccaccccaga gggaggccaa catctacatg gttcctcaga
     4021 acatcaagcc agcgctccag cgtaccgcca tcgagatcct ggcatggggc ctgcggaaca
     4081 tgaagagtta ccagctggcc aacatctcct cccccagcct cgtggtagag tgtgggggcc
     4141 agacggtgca gtcctgtgtc atcaggaacc tccggaagaa ccccaacttt gacatctgca
     4201 ccctcttcat ggaagtgatg ctgcccaggg aggagctcta ctgccccccc atcaccgtca
     4261 aggtcatcga taaccgccag tttggccgcc ggcctgtggt gggccagtgt accatccgct
     4321 ccctggagag cttcctgtgt gacccctact cggcggagag tccatcccca cagggtggcc
     4381 cagacgatgt gagcctactc agtcctgggg aagacgtgct catcgacatt gatgacaagg
     4441 agcccctcat ccccatccag gaggaagagt tcatcgattg gtggagcaaa ttctttgcct
     4501 ccatagggga gagggaaaag tgcggctcct acctggagaa ggattttgac accctgaagg
     4561 tctatgacac acagctggag aatgtggagg cctttgaggg cctgtctgac ttttgtaaca
     4621 ccttcaagct gtaccggggc aagacgcagg aggagacaga agatccatct gtgattggtg
     4681 aatttaaggg cctcttcaaa atttatcccc tcccagaaga cccagccatc cccatgcccc
     4741 caagacagtt ccaccagctg gccgcccagg gaccccagga gtgcttggtc cgtatctaca
     4801 ttgtccgagc atttggcctg cagcccaagg accccaatgg aaagtgtgat ccttacatca
     4861 agatctccat agggaagaaa tcagtgagtg accaggataa ctacatcccc tgcacgctgg
     4921 agcccgtatt tggaaagatg ttcgagctga cctgcactct gcctctggag aaggacctaa
     4981 agatcactct ctatgactat gacctcctct ccaaggacga aaagatcggt gagacggtcg
     5041 tcgacctgga gaacaggctg ctgtccaagt ttggggctcg ctgtggactc ccacagacct
     5101 actgtgtctc tggaccgaac cagtggcggg accagctccg cccctcccag ctcctccacc
     5161 tcttctgcca gcagcataga gtcaaggcac ctgtgtaccg gacagaccgt gtaatgtttc
     5221 aggataaaga atattccatt gaagagatag aggctggcag gatcccaaac ccacacctgg
     5281 gcccagtgga ggagcgtctg gctctgcatg tgcttcagca gcagggcctg gtcccggagc
     5341 acgtggagtc acggcccctc tacagccccc tgcagccaga catcgagcag gggaagctgc
     5401 agatgtgggt cgacctattt ccgaaggccc tggggcggcc tggacctccc ttcaacatca
     5461 ccccacggag agccagaagg tttttcctgc gttgtattat ctggaatacc agagatgtga
     5521 tcctggatga cctgagcctc acgggggaga agatgagcga catttatgtg aaaggttgga
     5581 tgattggctt tgaagaacac aagcaaaaga cagacgtgca ttatcgttcc ctgggaggtg
     5641 aaggcaactt caactggagg ttcattttcc ccttcgacta cctgccagct gagcaagtct
     5701 gtaccattgc caagaaggat gccttctgga ggctggacaa gactgagagc aaaatcccag
     5761 cacgagtggt gttccagatc tgggacaatg acaagttctc ctttgatgat tttctgggct
     5821 ccctgcagct cgatctcaac cgcatgccca agccagccaa gacagccaag aagtgctcct
     5881 tggaccagct ggatgatgct ttccacccag aatggtttgt gtcccttttt gagcagaaaa
     5941 cagtgaaggg ctggtggccc tgtgtagcag aagagggtga gaagaaaata ctggcgggca
     6001 agctggaaat gaccttggag attgtagcag agagtgagca tgaggagcgg cctgctggcc
     6061 agggccggga tgagcccaac atgaacccta agcttgagga cccaaggcgc cccgacacct
     6121 ccttcctgtg gtttacctcc ccatacaaga ccatgaagtt catcctgtgg cggcgtttcc
     6181 ggtgggccat catcctcttc atcatcctct tcatcctgct gctgttcctg gccatcttca
     6241 tctacgcctt cccgaactat gctgccatga agctggtgaa gcccttcagc tgaatcccaa
     6301 tggcgcgccg agcttggctc gagcatggtc atagctgttt cctgtgtgaa attgttatcc
     6361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6481 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7141 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt agaaaaactc
     7501 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     7561 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     7621 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     7681 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     7741 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     7801 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     7861 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     7921 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     7981 ctggaatgct gttttcccag ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     8041 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     8101 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     8161 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     8221 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     8281 cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca
     8341 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8401 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     8461 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     8521 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     8581 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     8641 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     8701 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     8761 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     8821 agggggatgt gctgcaaggc gattaagttg ggtaacgcca g
//
