LOCUS       sequence-174254-        8864 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 8864)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8864)
  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, 17
FEATURES             Location/Qualifiers
     source          1..8864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..33
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     25..42
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     26..42
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             736..747
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6335..6351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6335..6351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6348..6370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6359..6375
                     /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(6383..6389,6390..6407,6408..6413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6428..6449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6566..6583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6737..7325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6817..6836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7496..8311)
                     /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(7496..8311)
                     /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     8219..8238
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8312..8416)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8484..8502
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8540..8562)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8662..8681
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgac gcgtatgctg
       61 tgctgcctgc tggtgagggc cagcaacctc cccagtgcga agaaggaccg gcgcagcgac
      121 cctgtcgcaa gcctgacttt ccgaggggtg aagaagagaa ccaaagtcat caagaacagc
      181 gtgaaccctg tatggaatga gggatttgaa tgggacctca agggcatccc cctggaccag
      241 ggctctgagc ttcatgtggt ggtcaaagac catgagacga tggggaggaa caggttcctg
      301 ggggaagcca aggtcccact ccgagaggtc ctcgccaccc ctagtctgtc cgccagcttc
      361 aatgcccccc tgctggacac caagaagcag cccacagggg cctcgctggt cctgcaggtg
      421 tcctacacac cgctgcctgg agctgtgccc ctgttcccgc cccctactcc tctggagccc
      481 tccccgactc tgcctgacct ggatgtagtg gcagacacag gaggagagga agacacagag
      541 gaccagggac tcactggaga tgaggcggag ccattcctgg atcaaagcgg aggcccgggg
      601 gctcccacca ccccaaggaa actaccttca cgtcctccgc cccactaccc cgggatcaaa
      661 agaaagcgaa gtgcgcctac atctagaaag ctgctgtcag acaaaccgca ggatttccag
      721 atcagggtcc aggtgatcga ggggcgccag ctgccggggg tgaacatcaa gcctgtggtc
      781 aaggttaccg ctgcagggca gaccaagcgg acgcggatcc acaagggaaa cagcccactc
      841 ttcaatgaga ctcttttctt caacttgttt gactctcctg gggagctgtt tgatgagccc
      901 atctttatca cggtggtaga ctctcgttct ctcaggacag atgctctcct cggggagttc
      961 cggatggacg tgggcaccat ttacagagag ccccggcacg cctatctcag gaagtggctg
     1021 ctgctctcag accctgatga cttctctgct ggggccagag gctacctgaa aacaagcctt
     1081 tgtgtgctgg ggcctgggga cgaagcgcct ctggagagaa aagacccctc tgaagacaag
     1141 gaggacattg aaagcaacct gctccggccc acaggcgtag ccctgcgagg agcccacttc
     1201 tgcctgaagg tcttccgggc cgaggacttg ccgcagatgg acgatgccgt gatggacaac
     1261 gtgaaacaga tctttggctt cgagagtaac aagaagaact tggtggaccc ctttgtggag
     1321 gtcagctttg cggggaaaat gctgtgcagc aagatcttgg agaagacggc caaccctcag
     1381 tggaaccaga acatcacact gcctgccatg tttccctcca tgtgcgaaaa aatgaggatt
     1441 cgtatcatag actgggaccg cctgactcac aatgacatcg tggctaccac ctacctgagt
     1501 atgtcgaaaa tctctgcccc tggaggagaa atagaagagg agcctgcagg tgctgtcaag
     1561 ccttcgaaag cctcagactt ggatgactac ctgggcttcc tccccacttt tgggccctgc
     1621 tacatcaacc tctatggcag tcccagagag ttcacaggct tcccagaccc ctacacagag
     1681 ctcaacacag gcaaggggga aggtgtggct tatcgtggcc ggcttctgct ctccctggag
     1741 accaagctgg tggagcacag tgaacagaag gtggaggacc ttcctgcgga tgacatcctc
     1801 cgggtggaga agtaccttag gaggcgcaag tactccctgt ttgcggcctt ctactcagcc
     1861 accatgctgc aggatgtgga tgacgccatc cagtttgagg tcagcatcgg gaactacggg
     1921 aacaagttcg acatgacctg cctgccgctg gcctccacca ctcagtacag ccgtgcagtc
     1981 tttgacgggt gccactacta ctacctaccc tggggtaacg tgaaacccgt ggtggtgctg
     2041 tcatcctact gggaggacat cagccataga atcgagactc agaaccagct gcttgggatt
     2101 gctgaccggc tggaagctgg cctggagcag gtccacctgg ccctgaaggc gcagtgctcc
     2161 acggaggacg tggactcgct ggtggctcag ctgacggatg agctcatcgc aggctgcagc
     2221 cagcctctgg gtgacatcca tgagacaccc tctgccaccc acctggacca gtacctgtac
     2281 cagctgcgca cccatcacct gagccaaatc actgaggctg ccctggccct gaagctcggc
     2341 cacagtgagc tccctgcagc tctggagcag gcggaggact ggctcctgcg tctgcgtgcc
     2401 ctggcagagg agccccagaa cagcctgccg gacatcgtca tctggatgct gcagggagac
     2461 aagcgtgtgg cataccagcg ggtgcctgcc caccaagtcc tcttctcccg gcggggtgcc
     2521 aactactgtg gcaagaattg tgggaagcta cagacaatct ttctgaaata tccgatggag
     2581 aaggtgcctg gcgcccggat gccagtgcag atacgggtca agctgtggtt tgggctctct
     2641 gtggatgaga aggagttcaa ccagtttgct gaggggaagc tgtctgtctt tgctgaaacc
     2701 tatgagaacg agactaagtt ggcccttgtt gggaactggg gcacaacggg cctcacctac
     2761 cccaagtttt ctgacgtcac gggcaagatc aagctaccca aggacagctt ccgcccctcg
     2821 gccggctgga cctgggctgg agattggttc gtgtgcccgg agaagactct gctccatgac
     2881 atggacgccg gtcacctgag cttcgtggaa gaggtgtttg agaaccagac ccggcttccc
     2941 ggaggccagt ggatctacat gagtgacaac tacaccgatg tgaacgggga gaaggtgctt
     3001 cccaaggatg acattgagtg cccactgggc tggaagtggg aagatgagga atggtccaca
     3061 gacctcaacc gggctgtcga tgagcaaggc tgggagtata gcatcaccat ccccccggag
     3121 cggaagccga agcactgggt ccctgctgag aagatgtact acacacaccg acggcggcgc
     3181 tgggtgcgcc tgcgcaggag ggatctcagc caaatggaag cactgaaaag gcacaggcag
     3241 gcggaggcgg agggcgaggg ctgggagtac gcctctcttt ttggctggaa gttccacctc
     3301 gagtaccgca agacagatgc cttccgccgc cgccgctggc gccgtcgcat ggagccactg
     3361 gagaagacgg ggcctgcagc tgtgtttgcc cttgaggggg ccctgggcgg cgtgatggat
     3421 gacaagagtg aagattccat gtccgtctcc accttgagct tcggtgtgaa cagacccacg
     3481 atttcctgca tattcgacta tgggaaccgc taccatctac gctgctacat gtaccaggcc
     3541 cgggacctgg ctgcgatgga caaggactct ttttctgatc cctatgccat cgtctccttc
     3601 ctgcaccaga gccagaagac ggtggtggtg aagaacaccc ttaaccccac ctgggaccag
     3661 acgctcatct tctacgagat cgagatcttt ggcgagccgg ccacagttgc tgagcaaccg
     3721 cccagcattg tggtggagct gtacgaccat gacacttatg gtgcagacga gtttatgggt
     3781 cgctgcatct gtcaaccgag tctggaacgg atgccacggc tggcctggtt cccactgacg
     3841 aggggcagcc agccgtcggg ggagctgctg gcctcttttg agctcatcca gagagagaag
     3901 ccggccatcc accatattcc tggttttgag gtgcaggaga catcaaggat cctggatgag
     3961 tctgaggaca cagacctgcc ctacccacca ccccagaggg aggccaacat ctacatggtt
     4021 cctcagaaca tcaagccagc gctccagcgt accgccatcg agatcctggc atggggcctg
     4081 cggaacatga agagttacca gctggccaac atctcctccc ccagcctcgt ggtagagtgt
     4141 gggggccaga cggtgcagtc ctgtgtcatc aggaacctcc ggaagaaccc caactttgac
     4201 atctgcaccc tcttcatgga agtgatgctg cccagggagg agctctactg cccccccatc
     4261 accgtcaagg tcatcgataa ccgccagttt ggccgccggc ctgtggtggg ccagtgtacc
     4321 atccgctccc tggagagctt cctgtgtgac ccctactcgg cggagagtcc atccccacag
     4381 ggtggcccag acgatgtgag cctactcagt cctggggaag acgtgctcat cgacattgat
     4441 gacaaggagc ccctcatccc catccaggag gaagagttca tcgattggtg gagcaaattc
     4501 tttgcctcca taggggagag ggaaaagtgc ggctcctacc tggagaagga ttttgacacc
     4561 ctgaaggtct atgacacaca gctggagaat gtggaggcct ttgagggcct gtctgacttt
     4621 tgtaacacct tcaagctgta ccggggcaag acgcaggagg agacagaaga tccatctgtg
     4681 attggtgaat ttaagggcct cttcaaaatt tatcccctcc cagaagaccc agccatcccc
     4741 atgcccccaa gacagttcca ccagctggcc gcccagggac cccaggagtg cttggtccgt
     4801 atctacattg tccgagcatt tggcctgcag cccaaggacc ccaatggaaa gtgtgatcct
     4861 tacatcaaga tctccatagg gaagaaatca gtgagtgacc aggataacta catcccctgc
     4921 acgctggagc ccgtatttgg aaagatgttc gagctgacct gcactctgcc tctggagaag
     4981 gacctaaaga tcactctcta tgactatgac ctcctctcca aggacgaaaa gatcggtgag
     5041 acggtcgtcg acctggagaa caggctgctg tccaagtttg gggctcgctg tggactccca
     5101 cagacctact gtgtctctgg accgaaccag tggcgggacc agctccgccc ctcccagctc
     5161 ctccacctct tctgccagca gcatagagtc aaggcacctg tgtaccggac agaccgtgta
     5221 atgtttcagg ataaagaata ttccattgaa gagatagagg ctggcaggat cccaaaccca
     5281 cacctgggcc cagtggagga gcgtctggct ctgcatgtgc ttcagcagca gggcctggtc
     5341 ccggagcacg tggagtcacg gcccctctac agccccctgc agccagacat cgagcagggg
     5401 aagctgcaga tgtgggtcga cctatttccg aaggccctgg ggcggcctgg acctcccttc
     5461 aacatcaccc cacggagagc cagaaggttt ttcctgcgtt gtattatctg gaataccaga
     5521 gatgtgatcc tggatgacct gagcctcacg ggggagaaga tgagcgacat ttatgtgaaa
     5581 ggttggatga ttggctttga agaacacaag caaaagacag acgtgcatta tcgttccctg
     5641 ggaggtgaag gcaacttcaa ctggaggttc attttcccct tcgactacct gccagctgag
     5701 caagtctgta ccattgccaa gaaggatgcc ttctggaggc tggacaagac tgagagcaaa
     5761 atcccagcac gagtggtgtt ccagatctgg gacaatgaca agttctcctt tgatgatttt
     5821 ctgggctccc tgcagctcga tctcaaccgc atgcccaagc cagccaagac agccaagaag
     5881 tgctccttgg accagctgga tgatgctttc cacccagaat ggtttgtgtc cctttttgag
     5941 cagaaaacag tgaagggctg gtggccctgt gtagcagaag agggtgagaa gaaaatactg
     6001 gcgggcaagc tggaaatgac cttggagatt gtagcagaga gtgagcatga ggagcggcct
     6061 gctggccagg gccgggatga gcccaacatg aaccctaagc ttgaggaccc aaggcgcccc
     6121 gacacctcct tcctgtggtt tacctcccca tacaagacca tgaagttcat cctgtggcgg
     6181 cgtttccggt gggccatcat cctcttcatc atcctcttca tcctgctgct gttcctggcc
     6241 atcttcatct acgccttccc gaactatgct gccatgaagc tggtgaagcc cttcagctga
     6301 atcccaatgg cgcgccgagc ttggctcgag catggtcata gctgtttcct gtgtgaaatt
     6361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6481 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6541 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6601 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6661 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7141 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7321 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7441 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttaga
     7501 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     7561 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     7621 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     7681 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     7741 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     7801 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     7861 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     7921 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     7981 ctaatacctg gaatgctgtt ttcccaggga tcgcagtggt gagtaaccat gcatcatcag
     8041 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     8101 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     8161 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     8221 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     8281 agcaagacgt ttcccgttga atatggctca tactcttcct ttttcaatat tattgaagca
     8341 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8401 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     8461 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     8521 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     8581 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     8641 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     8701 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     8761 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     8821 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacg
//
