LOCUS       sequence-174220-        8924 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 8924)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8924)
  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, 40a
FEATURES             Location/Qualifiers
     source          1..8924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..46
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     38..55
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     39..55
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             746..757
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6408..6424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6408..6424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6421..6443)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6432..6448
                     /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(6456..6462,6463..6480,6481..6486))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6501..6522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6639..6656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6810..7398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6890..6909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7569..8384)
                     /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(7569..8384)
                     /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     8292..8311
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8385..8489)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8557..8575
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8613..8635)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8735..8754
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
       61 gacgcgtatg ctgagggtct tcatcctcta tgccgagaac gtccacacac ccgacaccga
      121 catcagcgat gcctactgct ccgcggtgtt tgcaggggtg aagaagagaa ccaaagtcat
      181 caagaacagc gtgaaccctg tatggaatga gggatttgaa tgggacctca agggcatccc
      241 cctggaccag ggctctgagc ttcatgtggt ggtcaaagac catgagacga tggggaggaa
      301 caggttcctg ggggaagcca aggtcccact ccgagaggtc ctcgccaccc ctagtctgtc
      361 cgccagcttc aatgcccccc tgctggacac caagaagcag cccacagggg cctcgctggt
      421 cctgcaggtg tcctacacac cgctgcctgg agctgtgccc ctgttcccgc cccctactcc
      481 tctggagccc tccccgactc tgcctgacct ggatgtagtg gcagacacag gaggagagga
      541 agacacagag gaccagggac tcactggaga tgaggcggag ccattcctgg atcaaagcgg
      601 aggcccgggg gctcccacca ccccaaggaa actaccttca cgtcctccgc cccactaccc
      661 cgggatcaaa agaaagcgaa gtgcgcctac atctagaaag ctgctgtcag acaaaccgca
      721 ggatttccag atcagggtcc aggtgatcga ggggcgccag ctgccggggg tgaacatcaa
      781 gcctgtggtc aaggttaccg ctgcagggca gaccaagcgg acgcggatcc acaagggaaa
      841 cagcccactc ttcaatgaga ctcttttctt caacttgttt gactctcctg gggagctgtt
      901 tgatgagccc atctttatca cggtggtaga ctctcgttct ctcaggacag atgctctcct
      961 cggggagttc cggatggacg tgggcaccat ttacagagag ccccggcacg cctatctcag
     1021 gaagtggctg ctgctctcag accctgatga cttctctgct ggggccagag gctacctgaa
     1081 aacaagcctt tgtgtgctgg ggcctgggga cgaagcgcct ctggagagaa aagacccctc
     1141 tgaagacaag gaggacattg aaagcaacct gctccggccc acaggcgtag ccctgcgagg
     1201 agcccacttc tgcctgaagg tcttccgggc cgaggacttg ccgcagatgg acgatgccgt
     1261 gatggacaac gtgaaacaga tctttggctt cgagagtaac aagaagaact tggtggaccc
     1321 ctttgtggag gtcagctttg cggggaaaat gctgtgcagc aagatcttgg agaagacggc
     1381 caaccctcag tggaaccaga acatcacact gcctgccatg tttccctcca tgtgcgaaaa
     1441 aatgaggatt cgtatcatag actgggaccg cctgactcac aatgacatcg tggctaccac
     1501 ctacctgagt atgtcgaaaa tctctgcccc tggaggagaa atagaagagg agcctgcagg
     1561 tgctgtcaag ccttcgaaag cctcagactt ggatgactac ctgggcttcc tccccacttt
     1621 tgggccctgc tacatcaacc tctatggcag tcccagagag ttcacaggct tcccagaccc
     1681 ctacacagag ctcaacacag gcaaggggga aggtgtggct tatcgtggcc ggcttctgct
     1741 ctccctggag accaagctgg tggagcacag tgaacagaag gtggaggacc ttcctgcgga
     1801 tgacatcctc cgggtggaga agtaccttag gaggcgcaag tactccctgt ttgcggcctt
     1861 ctactcagcc accatgctgc aggatgtgga tgacgccatc cagtttgagg tcagcatcgg
     1921 gaactacggg aacaagttcg acatgacctg cctgccgctg gcctccacca ctcagtacag
     1981 ccgtgcagtc tttgacgggt gccactacta ctacctaccc tggggtaacg tgaaacccgt
     2041 ggtggtgctg tcatcctact gggaggacat cagccataga atcgagactc agaaccagct
     2101 gcttgggatt gctgaccggc tggaagctgg cctggagcag gtccacctgg ccctgaaggc
     2161 gcagtgctcc acggaggacg tggactcgct ggtggctcag ctgacggatg agctcatcgc
     2221 aggctgcagc cagcctctgg gtgacatcca tgagacaccc tctgccaccc acctggacca
     2281 gtacctgtac cagctgcgca cccatcacct gagccaaatc actgaggctg ccctggccct
     2341 gaagctcggc cacagtgagc tccctgcagc tctggagcag gcggaggact ggctcctgcg
     2401 tctgcgtgcc ctggcagagg agccccagaa cagcctgccg gacatcgtca tctggatgct
     2461 gcagggagac aagcgtgtgg cataccagcg ggtgcctgcc caccaagtcc tcttctcccg
     2521 gcggggtgcc aactactgtg gcaagaattg tgggaagcta cagacaatct ttctgaaata
     2581 tccgatggag aaggtgcctg gcgcccggat gccagtgcag atacgggtca agctgtggtt
     2641 tgggctctct gtggatgaga aggagttcaa ccagtttgct gaggggaagc tgtctgtctt
     2701 tgctgaaacc tatgagaacg agactaagtt ggcccttgtt gggaactggg gcacaacggg
     2761 cctcacctac cccaagtttt ctgacgtcac gggcaagatc aagctaccca aggacagctt
     2821 ccgcccctcg gccggctgga cctgggctgg agattggttc gtgtgcccgg agaagactct
     2881 gctccatgac atggacgccg gtcacctgag cttcgtggaa gaggtgtttg agaaccagac
     2941 ccggcttccc ggaggccagt ggatctacat gagtgacaac tacaccgatg tgaacgggga
     3001 gaaggtgctt cccaaggatg acattgagtg cccactgggc tggaagtggg aagatgagga
     3061 atggtccaca gacctcaacc gggctgtcga tgagcaaggc tgggagtata gcatcaccat
     3121 ccccccggag cggaagccga agcactgggt ccctgctgag aagatgtact acacacaccg
     3181 acggcggcgc tgggtgcgcc tgcgcaggag ggatctcagc caaatggaag cactgaaaag
     3241 gcacaggcag gcggaggcgg agggcgaggg ctgggagtac gcctctcttt ttggctggaa
     3301 gttccacctc gagtaccgca agacagatgc cttccgccgc cgccgctggc gccgtcgcat
     3361 ggagccactg gagaagacgg ggcctgcagc tgtgtttgcc cttgaggggg ccctgggcgg
     3421 cgtgatggat gacaagagtg aagattccat gtccgtctcc accttgagct tcggtgtgaa
     3481 cagacccacg atttcctgca tattcgacta tgggaaccgc taccatctac gctgctacat
     3541 gtaccaggcc cgggacctgg ctgcgatgga caaggactct ttttctgatc cctatgccat
     3601 cgtctccttc ctgcaccaga gccagaagac ggtggtggtg aagaacaccc ttaaccccac
     3661 ctgggaccag acgctcatct tctacgagat cgagatcttt ggcgagccgg ccacagttgc
     3721 tgagcaaccg cccagcattg tggtggagct gtacgaccat gacacttatg gtgcagacga
     3781 gtttatgggt cgctgcatct gtcaaccgag tctggaacgg atgccacggc tggcctggtt
     3841 cccactgacg aggggcagcc agccgtcggg ggagctgctg gcctcttttg agctcatcca
     3901 gagagagaag ccggccatcc accatattcc tggttttgag gtgcaggaga catcaaggat
     3961 cctggatgag tctgaggaca cagacctgcc ctacccacca ccccagaggg aggccaacat
     4021 ctacatggtt cctcagaaca tcaagccagc gctccagcgt accgccatcg agatcctggc
     4081 atggggcctg cggaacatga agagttacca gctggccaac atctcctccc ccagcctcgt
     4141 ggtagagtgt gggggccaga cggtgcagtc ctgtgtcatc aggaacctcc ggaagaaccc
     4201 caactttgac atctgcaccc tcttcatgga agtgatgctg cccagggagg agctctactg
     4261 cccccccatc accgtcaagg tcatcgataa ccgccagttt ggccgccggc ctgtggtggg
     4321 ccagtgtacc atccgctccc tggagagctt cctgtgtgac ccctactcgg cggagagtcc
     4381 atccccacag ggtggcccag acgatgtgag cctactcagt cctggggaag acgtgctcat
     4441 cgacattgat gacaaggagc ccctcatccc catccagctt gcagacggtc tgtcgagctt
     4501 ggcccccact aacacggctt ctcctccatc cagtcctcat gaggaagagt tcatcgattg
     4561 gtggagcaaa ttctttgcct ccatagggga gagggaaaag tgcggctcct acctggagaa
     4621 ggattttgac accctgaagg tctatgacac acagctggag aatgtggagg cctttgaggg
     4681 cctgtctgac ttttgtaaca ccttcaagct gtaccggggc aagacgcagg aggagacaga
     4741 agatccatct gtgattggtg aatttaaggg cctcttcaaa atttatcccc tcccagaaga
     4801 cccagccatc cccatgcccc caagacagtt ccaccagctg gccgcccagg gaccccagga
     4861 gtgcttggtc cgtatctaca ttgtccgagc atttggcctg cagcccaagg accccaatgg
     4921 aaagtgtgat ccttacatca agatctccat agggaagaaa tcagtgagtg accaggataa
     4981 ctacatcccc tgcacgctgg agcccgtatt tggaaagatg ttcgagctga cctgcactct
     5041 gcctctggag aaggacctaa agatcactct ctatgactat gacctcctct ccaaggacga
     5101 aaagatcggt gagacggtcg tcgacctgga gaacaggctg ctgtccaagt ttggggctcg
     5161 ctgtggactc ccacagacct actgtgtctc tggaccgaac cagtggcggg accagctccg
     5221 cccctcccag ctcctccacc tcttctgcca gcagcataga gtcaaggcac ctgtgtaccg
     5281 gacagaccgt gtaatgtttc aggataaaga atattccatt gaagagatag aggctggcag
     5341 gatcccaaac ccacacctgg gcccagtgga ggagcgtctg gctctgcatg tgcttcagca
     5401 gcagggcctg gtcccggagc acgtggagtc acggcccctc tacagccccc tgcagccaga
     5461 catcgagcag gggaagctgc agatgtgggt cgacctattt ccgaaggccc tggggcggcc
     5521 tggacctccc ttcaacatca ccccacggag agccagaagg tttttcctgc gttgtattat
     5581 ctggaatacc agagatgtga tcctggatga cctgagcctc acgggggaga agatgagcga
     5641 catttatgtg aaaggttgga tgattggctt tgaagaacac aagcaaaaga cagacgtgca
     5701 ttatcgttcc ctgggaggtg aaggcaactt caactggagg ttcattttcc ccttcgacta
     5761 cctgccagct gagcaagtct gtaccattgc caagaaggat gccttctgga ggctggacaa
     5821 gactgagagc aaaatcccag cacgagtggt gttccagatc tgggacaatg acaagttctc
     5881 ctttgatgat tttctgggct ccctgcagct cgatctcaac cgcatgccca agccagccaa
     5941 gacagccaag aagtgctcct tggaccagct ggatgatgct ttccacccag aatggtttgt
     6001 gtcccttttt gagcagaaaa cagtgaaggg ctggtggccc tgtgtagcag aagagggtga
     6061 gaagaaaata ctggcgggca agctggaaat gaccttggag attgtagcag agagtgagca
     6121 tgaggagcgg cctgctggcc agggccggga tgagcccaac atgaacccta agcttgagga
     6181 cccaaggcgc cccgacacct ccttcctgtg gtttacctcc ccatacaaga ccatgaagtt
     6241 catcctgtgg cggcgtttcc ggtgggccat catcctcttc atcatcctct tcatcctgct
     6301 gctgttcctg gccatcttca tctacgcctt cccgaactat gctgccatga agctggtgaa
     6361 gcccttcagc tgaatcccaa tggcgcgccg agcttggctc gagcatggtc atagctgttt
     6421 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6481 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6541 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6601 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6661 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6721 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6781 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6841 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6901 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6961 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7021 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7081 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7141 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7201 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7261 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7321 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7381 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7441 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7501 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7561 tctgacagtt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     7621 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     7681 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     7741 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     7801 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     7861 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     7921 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     7981 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     8041 tcaggatatt cttctaatac ctggaatgct gttttcccag ggatcgcagt ggtgagtaac
     8101 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     8161 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     8221 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     8281 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     8341 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa
     8401 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8461 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8521 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     8581 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     8641 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     8701 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     8761 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     8821 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     8881 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gatt
//
