LOCUS       sequence-174261-        8822 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 8822)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8822)
  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
FEATURES             Location/Qualifiers
     source          1..8822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..43
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     35..52
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     36..52
                     /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(6303..6319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6303..6319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6316..6338)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6327..6343
                     /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(6351..6357,6358..6375,6376..6381))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6396..6417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6534..6551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6705..7293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6785..6804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7464..8279)
                     /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(7464..8279)
                     /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     8187..8206
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8280..8384)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8452..8470
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8508..8530)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8630..8649
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgac
       61 gcgtatgctg tgctgcctgc tggtgagggc cagcaacctc cccagtgcga agaaggaccg
      121 gcgcagcgac cctgtcgcaa gcctgacttt ccgaggggtg 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 atagaagtgg atgactacct
     1561 gggcttcctc cccacttttg ggccctgcta catcaacctc tatggcagtc ccagagagtt
     1621 cacaggcttc ccagacccct acacagagct caacacaggc aagggggaag gtgtggctta
     1681 tcgtggccgg cttctgctct ccctggagac caagctggtg gagcacagtg aacagaaggt
     1741 ggaggacctt cctgcggatg acatcctccg ggtggagaag taccttagga ggcgcaagta
     1801 ctccctgttt gcggccttct actcagccac catgctgcag gatgtggatg acgccatcca
     1861 gtttgaggtc agcatcggga actacgggaa caagttcgac atgacctgcc tgccgctggc
     1921 ctccaccact cagtacagcc gtgcagtctt tgacgggtgc cactactact acctaccctg
     1981 gggtaacgtg aaacccgtgg tggtgctgtc atcctactgg gaggacatca gccatagaat
     2041 cgagactcag aaccagctgc ttgggattgc tgaccggctg gaagctggcc tggagcaggt
     2101 ccacctggcc ctgaaggcgc agtgctccac ggaggacgtg gactcgctgg tggctcagct
     2161 gacggatgag ctcatcgcag gctgcagcca gcctctgggt gacatccatg agacaccctc
     2221 tgccacccac ctggaccagt acctgtacca gctgcgcacc catcacctga gccaaatcac
     2281 tgaggctgcc ctggccctga agctcggcca cagtgagctc cctgcagctc tggagcaggc
     2341 ggaggactgg ctcctgcgtc tgcgtgccct ggcagaggag ccccagaaca gcctgccgga
     2401 catcgtcatc tggatgctgc agggagacaa gcgtgtggca taccagcggg tgcctgccca
     2461 ccaagtcctc ttctcccggc ggggtgccaa ctactgtggc aagaattgtg ggaagctaca
     2521 gacaatcttt ctgaaatatc cgatggagaa ggtgcctggc gcccggatgc cagtgcagat
     2581 acgggtcaag ctgtggtttg ggctctctgt ggatgagaag gagttcaacc agtttgctga
     2641 ggggaagctg tctgtctttg ctgaaaccta tgagaacgag actaagttgg cccttgttgg
     2701 gaactggggc acaacgggcc tcacctaccc caagttttct gacgtcacgg gcaagatcaa
     2761 gctacccaag gacagcttcc gcccctcggc cggctggacc tgggctggag attggttcgt
     2821 gtgcccggag aagactctgc tccatgacat ggacgccggt cacctgagct tcgtggaaga
     2881 ggtgtttgag aaccagaccc ggcttcccgg aggccagtgg atctacatga gtgacaacta
     2941 caccgatgtg aacggggaga aggtgcttcc caaggatgac attgagtgcc cactgggctg
     3001 gaagtgggaa gatgaggaat ggtccacaga cctcaaccgg gctgtcgatg agcaaggctg
     3061 ggagtatagc atcaccatcc ccccggagcg gaagccgaag cactgggtcc ctgctgagaa
     3121 gatgtactac acacaccgac ggcggcgctg ggtgcgcctg cgcaggaggg atctcagcca
     3181 aatggaagca ctgaaaaggc acaggcaggc ggaggcggag ggcgagggct gggagtacgc
     3241 ctctcttttt ggctggaagt tccacctcga gtaccgcaag acagatgcct tccgccgccg
     3301 ccgctggcgc cgtcgcatgg agccactgga gaagacgggg cctgcagctg tgtttgccct
     3361 tgagggggcc ctgggcggcg tgatggatga caagagtgaa gattccatgt ccgtctccac
     3421 cttgagcttc ggtgtgaaca gacccacgat ttcctgcata ttcgactatg ggaaccgcta
     3481 ccatctacgc tgctacatgt accaggcccg ggacctggct gcgatggaca aggactcttt
     3541 ttctgatccc tatgccatcg tctccttcct gcaccagagc cagaagacgg tggtggtgaa
     3601 gaacaccctt aaccccacct gggaccagac gctcatcttc tacgagatcg agatctttgg
     3661 cgagccggcc acagttgctg agcaaccgcc cagcattgtg gtggagctgt acgaccatga
     3721 cacttatggt gcagacgagt ttatgggtcg ctgcatctgt caaccgagtc tggaacggat
     3781 gccacggctg gcctggttcc cactgacgag gggcagccag ccgtcggggg agctgctggc
     3841 ctcttttgag ctcatccaga gagagaagcc ggccatccac catattcctg gttttgaggt
     3901 gcaggagaca tcaaggatcc tggatgagtc tgaggacaca gacctgccct acccaccacc
     3961 ccagagggag gccaacatct acatggttcc tcagaacatc aagccagcgc tccagcgtac
     4021 cgccatcgag atcctggcat ggggcctgcg gaacatgaag agttaccagc tggccaacat
     4081 ctcctccccc agcctcgtgg tagagtgtgg gggccagacg gtgcagtcct gtgtcatcag
     4141 gaacctccgg aagaacccca actttgacat ctgcaccctc ttcatggaag tgatgctgcc
     4201 cagggaggag ctctactgcc cccccatcac cgtcaaggtc atcgataacc gccagtttgg
     4261 ccgccggcct gtggtgggcc agtgtaccat ccgctccctg gagagcttcc tgtgtgaccc
     4321 ctactcggcg gagagtccat ccccacaggg tggcccagac gatgtgagcc tactcagtcc
     4381 tggggaagac gtgctcatcg acattgatga caaggagccc ctcatcccca tccaggagga
     4441 agagttcatc gattggtgga gcaaattctt tgcctccata ggggagaggg aaaagtgcgg
     4501 ctcctacctg gagaaggatt ttgacaccct gaaggtctat gacacacagc tggagaatgt
     4561 ggaggccttt gagggcctgt ctgacttttg taacaccttc aagctgtacc ggggcaagac
     4621 gcaggaggag acagaagatc catctgtgat tggtgaattt aagggcctct tcaaaattta
     4681 tcccctccca gaagacccag ccatccccat gcccccaaga cagttccacc agctggccgc
     4741 ccagggaccc caggagtgct tggtccgtat ctacattgtc cgagcatttg gcctgcagcc
     4801 caaggacccc aatggaaagt gtgatcctta catcaagatc tccataggga agaaatcagt
     4861 gagtgaccag gataactaca tcccctgcac gctggagccc gtatttggaa agatgttcga
     4921 gctgacctgc actctgcctc tggagaagga cctaaagatc actctctatg actatgacct
     4981 cctctccaag gacgaaaaga tcggtgagac ggtcgtcgac ctggagaaca ggctgctgtc
     5041 caagtttggg gctcgctgtg gactcccaca gacctactgt gtctctggac cgaaccagtg
     5101 gcgggaccag ctccgcccct cccagctcct ccacctcttc tgccagcagc atagagtcaa
     5161 ggcacctgtg taccggacag accgtgtaat gtttcaggat aaagaatatt ccattgaaga
     5221 gatagaggct ggcaggatcc caaacccaca cctgggccca gtggaggagc gtctggctct
     5281 gcatgtgctt cagcagcagg gcctggtccc ggagcacgtg gagtcacggc ccctctacag
     5341 ccccctgcag ccagacatcg agcaggggaa gctgcagatg tgggtcgacc tatttccgaa
     5401 ggccctgggg cggcctggac ctcccttcaa catcacccca cggagagcca gaaggttttt
     5461 cctgcgttgt attatctgga ataccagaga tgtgatcctg gatgacctga gcctcacggg
     5521 ggagaagatg agcgacattt atgtgaaagg ttggatgatt ggctttgaag aacacaagca
     5581 aaagacagac gtgcattatc gttccctggg aggtgaaggc aacttcaact ggaggttcat
     5641 tttccccttc gactacctgc cagctgagca agtctgtacc attgccaaga aggatgcctt
     5701 ctggaggctg gacaagactg agagcaaaat cccagcacga gtggtgttcc agatctggga
     5761 caatgacaag ttctcctttg atgattttct gggctccctg cagctcgatc tcaaccgcat
     5821 gcccaagcca gccaagacag ccaagaagtg ctccttggac cagctggatg atgctttcca
     5881 cccagaatgg tttgtgtccc tttttgagca gaaaacagtg aagggctggt ggccctgtgt
     5941 agcagaagag ggtgagaaga aaatactggc gggcaagctg gaaatgacct tggagattgt
     6001 agcagagagt gagcatgagg agcggcctgc tggccagggc cgggatgagc ccaacatgaa
     6061 ccctaagctt gaggacccaa ggcgccccga cacctccttc ctgtggttta cctccccata
     6121 caagaccatg aagttcatcc tgtggcggcg tttccggtgg gccatcatcc tcttcatcat
     6181 cctcttcatc ctgctgctgt tcctggccat cttcatctac gccttcccga actatgctgc
     6241 catgaagctg gtgaagccct tcagctgaat cccaatggcg cgccgagctt ggctcgagca
     6301 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6361 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6421 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6481 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6541 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6601 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6661 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6721 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6781 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6841 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6901 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6961 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7021 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7081 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7141 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7201 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7261 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7321 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7381 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7441 tatgagtaaa cttggtctga cagttagaaa aactcatcga gcatcaaatg aaactgcaat
     7501 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     7561 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     7621 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     7681 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     7741 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     7801 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     7861 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     7921 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccagggatc
     7981 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     8041 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     8101 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     8161 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     8221 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     8281 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8341 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8401 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     8461 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     8521 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     8581 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     8641 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     8701 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     8761 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     8821 ag
//
