LOCUS       sequence-174256-        8885 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 8885)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8885)
  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, 40a
FEATURES             Location/Qualifiers
     source          1..8885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..42
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     34..51
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     35..51
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             745..756
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6365..6381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6365..6381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6378..6400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6389..6405
                     /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(6413..6419,6420..6437,6438..6443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6458..6479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6596..6613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6767..7355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6847..6866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7526..8341)
                     /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(7526..8341)
                     /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     8249..8268
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8342..8446)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8514..8532
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8570..8592)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8692..8711
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgacg
       61 cgtatgctgt gctgcctgct ggtgagggcc agcaacctcc ccagtgcgaa gaaggaccgg
      121 cgcagcgacc ctgtcgcaag cctgactttc cgaggggtga agaagagaac caaagtcatc
      181 aagaacagcg tgaaccctgt atggaatgag ggatttgaat gggacctcaa gggcatcccc
      241 ctggaccagg gctctgagct tcatgtggtg gtcaaagacc atgagacgat ggggaggaac
      301 aggttcctgg gggaagccaa ggtcccactc cgagaggtcc tcgccacccc tagtctgtcc
      361 gccagcttca atgcccccct gctggacacc aagaagcagc ccacaggggc ctcgctggtc
      421 ctgcaggtgt cctacacacc gctgcctgga gctgtgcccc tgttcccgcc ccctactcct
      481 ctggagccct ccccgactct gcctgacctg gatgtagtgg cagacacagg aggagaggaa
      541 gacacagagg accagggact cactggagat gaggcggagc cattcctgga tcaaagcgga
      601 ggcccggggg ctcccaccac cccaaggaaa ctaccttcac gtcctccgcc ccactacccc
      661 gggatcaaaa gaaagcgaag tgcgcctaca tctagaaagc tgctgtcaga caaaccgcag
      721 gatttccaga tcagggtcca ggtgatcgag gggcgccagc tgccgggggt gaacatcaag
      781 cctgtggtca aggttaccgc tgcagggcag accaagcgga cgcggatcca caagggaaac
      841 agcccactct tcaatgagac tcttttcttc aacttgtttg actctcctgg ggagctgttt
      901 gatgagccca tctttatcac ggtggtagac tctcgttctc tcaggacaga tgctctcctc
      961 ggggagttcc ggatggacgt gggcaccatt tacagagagc cccggcacgc ctatctcagg
     1021 aagtggctgc tgctctcaga ccctgatgac ttctctgctg gggccagagg ctacctgaaa
     1081 acaagccttt gtgtgctggg gcctggggac gaagcgcctc tggagagaaa agacccctct
     1141 gaagacaagg aggacattga aagcaacctg ctccggccca caggcgtagc cctgcgagga
     1201 gcccacttct gcctgaaggt cttccgggcc gaggacttgc cgcagatgga cgatgccgtg
     1261 atggacaacg tgaaacagat ctttggcttc gagagtaaca agaagaactt ggtggacccc
     1321 tttgtggagg tcagctttgc ggggaaaatg ctgtgcagca agatcttgga gaagacggcc
     1381 aaccctcagt ggaaccagaa catcacactg cctgccatgt ttccctccat gtgcgaaaaa
     1441 atgaggattc gtatcataga ctgggaccgc ctgactcaca atgacatcgt ggctaccacc
     1501 tacctgagta tgtcgaaaat ctctgcccct ggaggagaaa tagaagtgga tgactacctg
     1561 ggcttcctcc ccacttttgg gccctgctac atcaacctct atggcagtcc cagagagttc
     1621 acaggcttcc cagaccccta cacagagctc aacacaggca agggggaagg tgtggcttat
     1681 cgtggccggc ttctgctctc cctggagacc aagctggtgg agcacagtga acagaaggtg
     1741 gaggaccttc ctgcggatga catcctccgg gtggagaagt accttaggag gcgcaagtac
     1801 tccctgtttg cggccttcta ctcagccacc atgctgcagg atgtggatga cgccatccag
     1861 tttgaggtca gcatcgggaa ctacgggaac aagttcgaca tgacctgcct gccgctggcc
     1921 tccaccactc agtacagccg tgcagtcttt gacgggtgcc actactacta cctaccctgg
     1981 ggtaacgtga aacccgtggt ggtgctgtca tcctactggg aggacatcag ccatagaatc
     2041 gagactcaga accagctgct tgggattgct gaccggctgg aagctggcct ggagcaggtc
     2101 cacctggccc tgaaggcgca gtgctccacg gaggacgtgg actcgctggt ggctcagctg
     2161 acggatgagc tcatcgcagg ctgcagccag cctctgggtg acatccatga gacaccctct
     2221 gccacccacc tggaccagta cctgtaccag ctgcgcaccc atcacctgag ccaaatcact
     2281 gaggctgccc tggccctgaa gctcggccac agtgagctcc ctgcagctct ggagcaggcg
     2341 gaggactggc tcctgcgtct gcgtgccctg gcagaggagc cccagaacag cctgccggac
     2401 atcgtcatct ggatgctgca gggagacaag cgtgtggcat accagcgggt gcctgcccac
     2461 caagtcctct tctcccggcg gggtgccaac tactgtggca agaattgtgg gaagctacag
     2521 acaatctttc tgaaatatcc gatggagaag gtgcctggcg cccggatgcc agtgcagata
     2581 cgggtcaagc tgtggtttgg gctctctgtg gatgagaagg agttcaacca gtttgctgag
     2641 gggaagctgt ctgtctttgc tgaaacctat gagaacgaga ctaagttggc ccttgttggg
     2701 aactggggca caacgggcct cacctacccc aagttttctg acgtcacggg caagatcaag
     2761 ctacccaagg acagcttccg cccctcggcc ggctggacct gggctggaga ttggttcgtg
     2821 tgcccggaga agactctgct ccatgacatg gacgccggtc acctgagctt cgtggaagag
     2881 gtgtttgaga accagacccg gcttcccgga ggccagtgga tctacatgag tgacaactac
     2941 accgatgtga acggggagaa ggtgcttccc aaggatgaca ttgagtgccc actgggctgg
     3001 aagtgggaag atgaggaatg gtccacagac ctcaaccggg ctgtcgatga gcaaggctgg
     3061 gagtatagca tcaccatccc cccggagcgg aagccgaagc actgggtccc tgctgagaag
     3121 atgtactaca cacaccgacg gcggcgctgg gtgcgcctgc gcaggaggga tctcagccaa
     3181 atggaagcac tgaaaaggca caggcaggcg gaggcggagg gcgagggctg ggagtacgcc
     3241 tctctttttg gctggaagtt ccacctcgag taccgcaaga cagatgcctt ccgccgccgc
     3301 cgctggcgcc gtcgcatgga gccactggag aagacggggc ctgcagctgt gtttgccctt
     3361 gagggggccc tgggcggcgt gatggatgac aagagtgaag attccatgtc cgtctccacc
     3421 ttgagcttcg gtgtgaacag acccacgatt tcctgcatat tcgactatgg gaaccgctac
     3481 catctacgct gctacatgta ccaggcccgg gacctggctg cgatggacaa ggactctttt
     3541 tctgatccct atgccatcgt ctccttcctg caccagagcc agaagacggt ggtggtgaag
     3601 aacaccctta accccacctg ggaccagacg ctcatcttct acgagatcga gatctttggc
     3661 gagccggcca cagttgctga gcaaccgccc agcattgtgg tggagctgta cgaccatgac
     3721 acttatggtg cagacgagtt tatgggtcgc tgcatctgtc aaccgagtct ggaacggatg
     3781 ccacggctgg cctggttccc actgacgagg ggcagccagc cgtcggggga gctgctggcc
     3841 tcttttgagc tcatccagag agagaagccg gccatccacc atattcctgg ttttgaggtg
     3901 caggagacat caaggatcct ggatgagtct gaggacacag acctgcccta cccaccaccc
     3961 cagagggagg ccaacatcta catggttcct cagaacatca agccagcgct ccagcgtacc
     4021 gccatcgaga tcctggcatg gggcctgcgg aacatgaaga gttaccagct ggccaacatc
     4081 tcctccccca gcctcgtggt agagtgtggg ggccagacgg tgcagtcctg tgtcatcagg
     4141 aacctccgga agaaccccaa ctttgacatc tgcaccctct tcatggaagt gatgctgccc
     4201 agggaggagc tctactgccc ccccatcacc gtcaaggtca tcgataaccg ccagtttggc
     4261 cgccggcctg tggtgggcca gtgtaccatc cgctccctgg agagcttcct gtgtgacccc
     4321 tactcggcgg agagtccatc cccacagggt ggcccagacg atgtgagcct actcagtcct
     4381 ggggaagacg tgctcatcga cattgatgac aaggagcccc tcatccccat ccagcttgca
     4441 gacggtctgt cgagcttggc ccccactaac acggcttctc ctccatccag tcctcatgag
     4501 gaagagttca tcgattggtg gagcaaattc tttgcctcca taggggagag ggaaaagtgc
     4561 ggctcctacc tggagaagga ttttgacacc ctgaaggtct atgacacaca gctggagaat
     4621 gtggaggcct ttgagggcct gtctgacttt tgtaacacct tcaagctgta ccggggcaag
     4681 acgcaggagg agacagaaga tccatctgtg attggtgaat ttaagggcct cttcaaaatt
     4741 tatcccctcc cagaagaccc agccatcccc atgcccccaa gacagttcca ccagctggcc
     4801 gcccagggac cccaggagtg cttggtccgt atctacattg tccgagcatt tggcctgcag
     4861 cccaaggacc ccaatggaaa gtgtgatcct tacatcaaga tctccatagg gaagaaatca
     4921 gtgagtgacc aggataacta catcccctgc acgctggagc ccgtatttgg aaagatgttc
     4981 gagctgacct gcactctgcc tctggagaag gacctaaaga tcactctcta tgactatgac
     5041 ctcctctcca aggacgaaaa gatcggtgag acggtcgtcg acctggagaa caggctgctg
     5101 tccaagtttg gggctcgctg tggactccca cagacctact gtgtctctgg accgaaccag
     5161 tggcgggacc agctccgccc ctcccagctc ctccacctct tctgccagca gcatagagtc
     5221 aaggcacctg tgtaccggac agaccgtgta atgtttcagg ataaagaata ttccattgaa
     5281 gagatagagg ctggcaggat cccaaaccca cacctgggcc cagtggagga gcgtctggct
     5341 ctgcatgtgc ttcagcagca gggcctggtc ccggagcacg tggagtcacg gcccctctac
     5401 agccccctgc agccagacat cgagcagggg aagctgcaga tgtgggtcga cctatttccg
     5461 aaggccctgg ggcggcctgg acctcccttc aacatcaccc cacggagagc cagaaggttt
     5521 ttcctgcgtt gtattatctg gaataccaga gatgtgatcc tggatgacct gagcctcacg
     5581 ggggagaaga tgagcgacat ttatgtgaaa ggttggatga ttggctttga agaacacaag
     5641 caaaagacag acgtgcatta tcgttccctg ggaggtgaag gcaacttcaa ctggaggttc
     5701 attttcccct tcgactacct gccagctgag caagtctgta ccattgccaa gaaggatgcc
     5761 ttctggaggc tggacaagac tgagagcaaa atcccagcac gagtggtgtt ccagatctgg
     5821 gacaatgaca agttctcctt tgatgatttt ctgggctccc tgcagctcga tctcaaccgc
     5881 atgcccaagc cagccaagac agccaagaag tgctccttgg accagctgga tgatgctttc
     5941 cacccagaat ggtttgtgtc cctttttgag cagaaaacag tgaagggctg gtggccctgt
     6001 gtagcagaag agggtgagaa gaaaatactg gcgggcaagc tggaaatgac cttggagatt
     6061 gtagcagaga gtgagcatga ggagcggcct gctggccagg gccgggatga gcccaacatg
     6121 aaccctaagc ttgaggaccc aaggcgcccc gacacctcct tcctgtggtt tacctcccca
     6181 tacaagacca tgaagttcat cctgtggcgg cgtttccggt gggccatcat cctcttcatc
     6241 atcctcttca tcctgctgct gttcctggcc atcttcatct acgccttccc gaactatgct
     6301 gccatgaagc tggtgaagcc cttcagctga atcccaatgg cgcgccgagc ttggctcgag
     6361 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6421 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6481 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6541 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6601 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6661 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6721 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6781 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6841 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6901 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6961 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7021 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7081 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7141 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7201 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7261 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7321 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7381 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7441 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7501 tatatgagta aacttggtct gacagttaga aaaactcatc gagcatcaaa tgaaactgca
     7561 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     7621 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     7681 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     7741 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     7801 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     7861 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     7921 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     7981 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccaggga
     8041 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     8101 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     8161 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     8221 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     8281 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     8341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8461 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8521 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8581 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8641 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8701 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8761 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8821 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8881 taagt
//
