LOCUS       sequence-174218-        8920 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 8920)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8920)
  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
FEATURES             Location/Qualifiers
     source          1..8920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..88
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     80..97
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     81..97
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             889..900
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6446..6462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6446..6462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6459..6481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6470..6486
                     /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(6494..6500,6501..6518,6519..6524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6539..6560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6677..6694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6848..7436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6928..6947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7607..8422)
                     /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(7607..8422)
                     /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     8330..8349
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8423..8527)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8595..8613
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8651..8673)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8773..8792
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
       61 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgacgcgta ttgggatatg
      121 ctgagggtct tcatcctcta tgccgagaac gtccacacac ccgacaccga catcagcgat
      181 gcctactgct ccgcggtgtt tgcaggggtg aagaagagaa ccaaagtcat caagaacagc
      241 gtgaaccctg tatggaatga gggatttgaa tgggacctca agggcatccc cctggaccag
      301 ggctctgagc ttcatgtggt ggtcaaagac catgagacga tggggaggaa caggttcctg
      361 ggggaagcca aggtcccact ccgagaggtc ctcgccaccc ctagtctgtc cgccagcttc
      421 aatgcccccc tgctggacac caagaagcag cccacagggg cctcgctggt cctgcaggtg
      481 tcctacacac cgctgcctgg agctgtgccc ctgttcccgc cccctactcc tctggagccc
      541 tccccgactc tgcctgacct ggatgtagtg gcaggcgggg gacagagccg ggccgagact
      601 tggtccctgc tcagtgacag caccatggac acgagatact ctggaaagaa gtggccggcc
      661 cccacggaca caggaggaga ggaagacaca gaggaccagg gactcactgg agatgaggcg
      721 gagccattcc tggatcaaag cggaggcccg ggggctccca ccaccccaag gaaactacct
      781 tcacgtcctc cgccccacta ccccgggatc aaaagaaagc gaagtgcgcc tacatctaga
      841 aagctgctgt cagacaaacc gcaggatttc cagatcaggg tccaggtgat cgaggggcgc
      901 cagctgccgg gggtgaacat caagcctgtg gtcaaggtta ccgctgcagg gcagaccaag
      961 cggacgcgga tccacaaggg aaacagccca ctcttcaatg agactctttt cttcaacttg
     1021 tttgactctc ctggggagct gtttgatgag cccatcttta tcacggtggt agactctcgt
     1081 tctctcagga cagatgctct cctcggggag ttccggatgg acgtgggcac catttacaga
     1141 gagccccggc acgcctatct caggaagtgg ctgctgctct cagaccctga tgacttctct
     1201 gctggggcca gaggctacct gaaaacaagc ctttgtgtgc tggggcctgg ggacgaagcg
     1261 cctctggaga gaaaagaccc ctctgaagac aaggaggaca ttgaaagcaa cctgctccgg
     1321 cccacaggcg tagccctgcg aggagcccac ttctgcctga aggtcttccg ggccgaggac
     1381 ttgccgcaga tggacgatgc cgtgatggac aacgtgaaac agatctttgg cttcgagagt
     1441 aacaagaaga acttggtgga cccctttgtg gaggtcagct ttgcggggaa aatgctgtgc
     1501 agcaagatct tggagaagac ggccaaccct cagtggaacc agaacatcac actgcctgcc
     1561 atgtttccct ccatgtgcga aaaaatgagg attcgtatca tagactggga ccgcctgact
     1621 cacaatgaca tcgtggctac cacctacctg agtatgtcga aaatctctgc ccctggagga
     1681 gaaatagaag tggatgacta cctgggcttc ctccccactt ttgggccctg ctacatcaac
     1741 ctctatggca gtcccagaga gttcacaggc ttcccagacc cctacacaga gctcaacaca
     1801 ggcaaggggg aaggtgtggc ttatcgtggc cggcttctgc tctccctgga gaccaagctg
     1861 gtggagcaca gtgaacagaa ggtggaggac cttcctgcgg atgacatcct ccgggtggag
     1921 aagtacctta ggaggcgcaa gtactccctg tttgcggcct tctactcagc caccatgctg
     1981 caggatgtgg atgacgccat ccagtttgag gtcagcatcg ggaactacgg gaacaagttc
     2041 gacatgacct gcctgccgct ggcctccacc actcagtaca gccgtgcagt ctttgacggg
     2101 tgccactact actacctacc ctggggtaac gtgaaacccg tggtggtgct gtcatcctac
     2161 tgggaggaca tcagccatag aatcgagact cagaaccagc tgcttgggat tgctgaccgg
     2221 ctggaagctg gcctggagca ggtccacctg gccctgaagg cgcagtgctc cacggaggac
     2281 gtggactcgc tggtggctca gctgacggat gagctcatcg caggctgcag ccagcctctg
     2341 ggtgacatcc atgagacacc ctctgccacc cacctggacc agtacctgta ccagctgcgc
     2401 acccatcacc tgagccaaat cactgaggct gccctggccc tgaagctcgg ccacagtgag
     2461 ctccctgcag ctctggagca ggcggaggac tggctcctgc gtctgcgtgc cctggcagag
     2521 gagccccaga acagcctgcc ggacatcgtc atctggatgc tgcagggaga caagcgtgtg
     2581 gcataccagc gggtgcctgc ccaccaagtc ctcttctccc ggcggggtgc caactactgt
     2641 ggcaagaatt gtgggaagct acagacaatc tttctgaaat atccgatgga gaaggtgcct
     2701 ggcgcccgga tgccagtgca gatacgggtc aagctgtggt ttgggctctc tgtggatgag
     2761 aaggagttca accagtttgc tgaggggaag ctgtctgtct ttgctgaaac ctatgagaac
     2821 gagactaagt tggcccttgt tgggaactgg ggcacaacgg gcctcaccta ccccaagttt
     2881 tctgacgtca cgggcaagat caagctaccc aaggacagct tccgcccctc ggccggctgg
     2941 acctgggctg gagattggtt cgtgtgcccg gagaagactc tgctccatga catggacgcc
     3001 ggtcacctga gcttcgtgga agaggtgttt gagaaccaga cccggcttcc cggaggccag
     3061 tggatctaca tgagtgacaa ctacaccgat gtgaacgggg agaaggtgct tcccaaggat
     3121 gacattgagt gcccactggg ctggaagtgg gaagatgagg aatggtccac agacctcaac
     3181 cgggctgtcg atgagcaagg ctgggagtat agcatcacca tccccccgga gcggaagccg
     3241 aagcactggg tccctgctga gaagatgtac tacacacacc gacggcggcg ctgggtgcgc
     3301 ctgcgcagga gggatctcag ccaaatggaa gcactgaaaa ggcacaggca ggcggaggcg
     3361 gagggcgagg gctgggagta cgcctctctt tttggctgga agttccacct cgagtaccgc
     3421 aagacagatg ccttccgccg ccgccgctgg cgccgtcgca tggagccact ggagaagacg
     3481 gggcctgcag ctgtgtttgc ccttgagggg gccctgggcg gcgtgatgga tgacaagagt
     3541 gaagattcca tgtccgtctc caccttgagc ttcggtgtga acagacccac gatttcctgc
     3601 atattcgact atgggaaccg ctaccatcta cgctgctaca tgtaccaggc ccgggacctg
     3661 gctgcgatgg acaaggactc tttttctgat ccctatgcca tcgtctcctt cctgcaccag
     3721 agccagaaga cggtggtggt gaagaacacc cttaacccca cctgggacca gacgctcatc
     3781 ttctacgaga tcgagatctt tggcgagccg gccacagttg ctgagcaacc gcccagcatt
     3841 gtggtggagc tgtacgacca tgacacttat ggtgcagacg agtttatggg tcgctgcatc
     3901 tgtcaaccga gtctggaacg gatgccacgg ctggcctggt tcccactgac gaggggcagc
     3961 cagccgtcgg gggagctgct ggcctctttt gagctcatcc agagagagaa gccggccatc
     4021 caccatattc ctggttttga ggtgcaggag acatcaagga tcctggatga gtctgaggac
     4081 acagacctgc cctacccacc accccagagg gaggccaaca tctacatggt tcctcagaac
     4141 atcaagccag cgctccagcg taccgccatc gagatcctgg catggggcct gcggaacatg
     4201 aagagttacc agctggccaa catctcctcc cccagcctcg tggtagagtg tgggggccag
     4261 acggtgcagt cctgtgtcat caggaacctc cggaagaacc ccaactttga catctgcacc
     4321 ctcttcatgg aagtgatgct gcccagggag gagctctact gcccccccat caccgtcaag
     4381 gtcatcgata accgccagtt tggccgccgg cctgtggtgg gccagtgtac catccgctcc
     4441 ctggagagct tcctgtgtga cccctactcg gcggagagtc catccccaca gggtggccca
     4501 gacgatgtga gcctactcag tcctggggaa gacgtgctca tcgacattga tgacaaggag
     4561 cccctcatcc ccatccagga ggaagagttc atcgattggt ggagcaaatt ctttgcctcc
     4621 ataggggaga gggaaaagtg cggctcctac ctggagaagg attttgacac cctgaaggtc
     4681 tatgacacac agctggagaa tgtggaggcc tttgagggcc tgtctgactt ttgtaacacc
     4741 ttcaagctgt accggggcaa gacgcaggag gagacagaag atccatctgt gattggtgaa
     4801 tttaagggcc tcttcaaaat ttatcccctc ccagaagacc cagccatccc catgccccca
     4861 agacagttcc accagctggc cgcccaggga ccccaggagt gcttggtccg tatctacatt
     4921 gtccgagcat ttggcctgca gcccaaggac cccaatggaa agtgtgatcc ttacatcaag
     4981 atctccatag ggaagaaatc agtgagtgac caggataact acatcccctg cacgctggag
     5041 cccgtatttg gaaagatgtt cgagctgacc tgcactctgc ctctggagaa ggacctaaag
     5101 atcactctct atgactatga cctcctctcc aaggacgaaa agatcggtga gacggtcgtc
     5161 gacctggaga acaggctgct gtccaagttt ggggctcgct gtggactccc acagacctac
     5221 tgtgtctctg gaccgaacca gtggcgggac cagctccgcc cctcccagct cctccacctc
     5281 ttctgccagc agcatagagt caaggcacct gtgtaccgga cagaccgtgt aatgtttcag
     5341 gataaagaat attccattga agagatagag gctggcagga tcccaaaccc acacctgggc
     5401 ccagtggagg agcgtctggc tctgcatgtg cttcagcagc agggcctggt cccggagcac
     5461 gtggagtcac ggcccctcta cagccccctg cagccagaca tcgagcaggg gaagctgcag
     5521 atgtgggtcg acctatttcc gaaggccctg gggcggcctg gacctccctt caacatcacc
     5581 ccacggagag ccagaaggtt tttcctgcgt tgtattatct ggaataccag agatgtgatc
     5641 ctggatgacc tgagcctcac gggggagaag atgagcgaca tttatgtgaa aggttggatg
     5701 attggctttg aagaacacaa gcaaaagaca gacgtgcatt atcgttccct gggaggtgaa
     5761 ggcaacttca actggaggtt cattttcccc ttcgactacc tgccagctga gcaagtctgt
     5821 accattgcca agaaggatgc cttctggagg ctggacaaga ctgagagcaa aatcccagca
     5881 cgagtggtgt tccagatctg ggacaatgac aagttctcct ttgatgattt tctgggctcc
     5941 ctgcagctcg atctcaaccg catgcccaag ccagccaaga cagccaagaa gtgctccttg
     6001 gaccagctgg atgatgcttt ccacccagaa tggtttgtgt ccctttttga gcagaaaaca
     6061 gtgaagggct ggtggccctg tgtagcagaa gagggtgaga agaaaatact ggcgggcaag
     6121 ctggaaatga ccttggagat tgtagcagag agtgagcatg aggagcggcc tgctggccag
     6181 ggccgggatg agcccaacat gaaccctaag cttgaggacc caaggcgccc cgacacctcc
     6241 ttcctgtggt ttacctcccc atacaagacc atgaagttca tcctgtggcg gcgtttccgg
     6301 tgggccatca tcctcttcat catcctcttc atcctgctgc tgttcctggc catcttcatc
     6361 tacgccttcc cgaactatgc tgccatgaag ctggtgaagc ccttcagctg aatcccaatg
     6421 gcgcgccgag cttggctcga gcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6481 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6541 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6601 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7261 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7561 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat
     7621 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     7681 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     7741 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     7801 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     7861 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     7921 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     7981 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     8041 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     8101 ggaatgctgt tttcccaggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     8161 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     8221 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     8281 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     8341 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     8401 tttcccgttg aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg
     8461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8521 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8581 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     8641 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     8701 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     8761 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     8821 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     8881 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
//
