LOCUS       sequence-96283-e        8533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 34 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8533)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8533)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: DYSF-Exon34-del
FEATURES             Location/Qualifiers
     source          1..8533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             897..908
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      6348..6357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6357..6373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6357..6373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6370..6392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6381..6397
                     /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(6405..6411,6412..6429,6430..6435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6450..6471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6588..6605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6759..7347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6839..6858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7518..8333)
                     /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(7518..8333)
                     /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     8241..8260
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8334..8438)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8506..8524
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 acgaattcga tatcgttaac ggtaccacta gtacgcgtat gctgagggtc ttcatcctct
      241 atgccgagaa cgtccacaca cccgacaccg acatcagcga tgcctactgc tccgcggtgt
      301 ttgcaggggt gaagaagaga accaaagtca tcaagaacag cgtgaaccct gtatggaatg
      361 agggatttga atgggacctc aagggcatcc ccctggacca gggctctgag cttcatgtgg
      421 tggtcaaaga ccatgagacg atggggagga acaggttcct gggggaagcc aaggtcccac
      481 tccgagaggt cctcgccacc cctagtctgt ccgccagctt caatgccccc ctgctggaca
      541 ccaagaagca gcccacaggg gcctcgctgg tcctgcaggt gtcctacaca ccgctgcctg
      601 gagctgtgcc cctgttcccg ccccctactc ctctggagcc ctccccgact ctgcctgacc
      661 tggatgtagt ggcagacaca ggaggagagg aagacacaga ggaccaggga ctcactggag
      721 atgaggcgga gccattcctg gatcaaagcg gaggcccggg ggctcccacc accccaagga
      781 aactaccttc acgtcctccg ccccactacc ccgggatcaa aagaaagcga agtgcgccta
      841 catctagaaa gctgctgtca gacaaaccgc aggatttcca gatcagggtc caggtgatcg
      901 aggggcgcca gctgccgggg gtgaacatca agcctgtggt caaggttacc gctgcagggc
      961 agaccaagcg gacgcggatc cacaagggaa acagcccact cttcaatgag actcttttct
     1021 tcaacttgtt tgactctcct ggggagctgt ttgatgagcc catctttatc acggtggtag
     1081 actctcgttc tctcaggaca gatgctctcc tcggggagtt ccggatggac gtgggcacca
     1141 tttacagaga gccccggcac gcctatctca ggaagtggct gctgctctca gaccctgatg
     1201 acttctctgc tggggccaga ggctacctga aaacaagcct ttgtgtgctg gggcctgggg
     1261 acgaagcgcc tctggagaga aaagacccct ctgaagacaa ggaggacatt gaaagcaacc
     1321 tgctccggcc cacaggcgta gccctgcgag gagcccactt ctgcctgaag gtcttccggg
     1381 ccgaggactt gccgcagatg gacgatgccg tgatggacaa cgtgaaacag atctttggct
     1441 tcgagagtaa caagaagaac ttggtggacc cctttgtgga ggtcagcttt gcggggaaaa
     1501 tgctgtgcag caagatcttg gagaagacgg ccaaccctca gtggaaccag aacatcacac
     1561 tgcctgccat gtttccctcc atgtgcgaaa aaatgaggat tcgtatcata gactgggacc
     1621 gcctgactca caatgacatc gtggctacca cctacctgag tatgtcgaaa atctctgccc
     1681 ctggaggaga aatagaagag gagcctgcag gtgctgtcaa gccttcgaaa gcctcagact
     1741 tggatgacta cctgggcttc ctccccactt ttgggccctg ctacatcaac ctctatggca
     1801 gtcccagaga gttcacaggc ttcccagacc cctacacaga gctcaacaca ggcaaggggg
     1861 aaggtgtggc ttatcgtggc cggcttctgc tctccctgga gaccaagctg gtggagcaca
     1921 gtgaacagaa ggtggaggac cttcctgcgg atgacatcct ccgggtggag aagtacctta
     1981 ggaggcgcaa gtactccctg tttgcggcct tctactcagc caccatgctg caggatgtgg
     2041 atgatgccat ccagtttgag gtcagcatcg ggaactacgg gaacaagttc gacatgacct
     2101 gcctgccgct ggcctccacc actcagtaca gccgtgcagt ctttgacggg tgccactact
     2161 actacctacc ctggggtaac gtgaaacctg tggtggtgct gtcatcctac tgggaggaca
     2221 tcagccatag aatcgagact cagaaccagc tgcttgggat tgctgaccgg ctggaagctg
     2281 gcctggagca ggtccacctg gccctgaagg cgcagtgctc cacggaggac gtggactcgc
     2341 tggtggctca gctgacggat gagctcatcg caggctgcag ccagcctctg ggtgacatcc
     2401 atgagacacc ctctgccacc cacctggacc agtacctgta ccagctgcgc acccatcacc
     2461 tgagccaaat cactgaggct gccctggccc tgaagctcgg ccacagtgag ctccctgcag
     2521 ctctggagca ggcggaggac tggctcctgc gtctgcgtgc cctggcagag gagccccaga
     2581 acagcctgcc ggacatcgtc atctggatgc tgcagggaga caagcgtgtg gcataccagc
     2641 gggtgcccgc ccaccaagtc ctcttctccc ggcggggtgc caactactgt ggcaagaatt
     2701 gtgggaagct acagacaatc tttctgaaat atccgatgga gaaggtgcct ggcgcccgga
     2761 tgccagtgca gatacgggtc aagctgtggt ttgggctctc tgtggatgag aaggagttca
     2821 accagtttgc tgaggggaag ctgtctgtct ttgctgaaac ctatgagaac gagactaagt
     2881 tggcccttgt tgggaactgg ggcacaacgg gcctcaccta ccccaagttt tctgacgtca
     2941 cgggcaagat caagctaccc aaggacagct tccgcccctc ggccggctgg acctgggctg
     3001 gagattggtt cgtgtgtccg gagaagactc tgctccatga catggacgcc ggtcacctga
     3061 gcttcgtgga agaggtgttt gagaaccaga cccggcttcc cggaggccag tggatctaca
     3121 tgagtgacaa ctacaccgat gtgaacgggg agaaggtgct tcccaaggat gacattgagt
     3181 gcccactggg ctggaagtgg gaagatgagg aatggtccac agacctcaac cgggctgtcg
     3241 atgagcaagg ctgggagtat agcatcacca tccccccgga gcggaagccg aagcactggg
     3301 tccctgctga gaagatgtac tacacacacc gacggcggcg ctgggtgcgc ctgcgcagga
     3361 gggatctcag ccaaatggaa gcactgaaaa ggcacaggca ggcggaggcg gagggcgagg
     3421 gctgggagta cgcctctctt tttggctgga agttccacct cgagtaccgc aagacagatg
     3481 ccttccgccg ccgccgctgg cgccgtcgca tggagccact ggagaagacg gggcctgcag
     3541 ctgtgtttgc ccttgagggg gccctgggcg gcgtgatgga tgacaagagt gaagattcca
     3601 tgtccgtctc caccttgagc ttcggtgtga acagacccac gatttcctgc atattcgact
     3661 atgggaaccg ctaccatcta cgctgctaca tgtaccaggc ccgggacctg gctgcgatgg
     3721 acaaggactc tttttctgat ccctatgcca tcgtctcctt cctgcaccag agccagaaga
     3781 cggtggtggt gaagaacacc cttaacccca cctgggacca gacgctcatc ttctacgaga
     3841 tcgagatctt tggcgagccg gccacagttg ctgagcaacc gcccagcatt gtggtggagc
     3901 tgtacgacca tgacacttat ccggccatcc accatattcc tggttttgag gtgcaggaga
     3961 catcaaggat cctggatgag tctgaggaca cagacctgcc ctacccacca ccccagaggg
     4021 aggccaacat ctacatggtt cctcagaaca tcaagccagc gctccagcgt accgccatcg
     4081 agatcctggc atggggcctg cggaacatga agagttacca gctggccaac atctcctccc
     4141 ccagcctcgt ggtagagtgt gggggccaga cggtgcagtc ctgtgtcatc aggaacctcc
     4201 ggaagaaccc caactttgac atctgcaccc tcttcatgga agtgatgctg cccagggagg
     4261 agctctactg cccccccatc accgtcaagg tcatcgataa ccgccagttt ggccgccggc
     4321 ctgtggtggg ccagtgtacc atccgctccc tggagagctt cctgtgtgac ccctactcgg
     4381 cggagagtcc atccccacag ggtggcccag acgatgtgag cctactcagt cctggggaag
     4441 acgtgctcat cgacattgat gacaaggagc ccctcatccc catccaggag gaagagttca
     4501 tcgattggtg gagcaaattc tttgcctcca taggggagag ggaaaagtgc ggctcctacc
     4561 tggagaagga ttttgacacc ctgaaggtct atgacacaca gctggagaat gtggaggcct
     4621 ttgagggcct gtctgacttt tgtaacacct tcaagctgta ccggggcaag acgcaggagg
     4681 agacagaaga tccatctgtg attggtgaat ttaagggcct cttcaaaatt tatcccctcc
     4741 cagaagaccc agccatcccc atgcccccaa gacagttcca ccagctggcc gcccagggac
     4801 cccaggagtg cttggtccgt atctacattg tccgagcatt tggcctgcag cccaaggacc
     4861 ccaatggaaa gtgtgatcct tacatcaaga tctccatagg gaagaaatca gtgagtgacc
     4921 aggataacta catcccctgc acgctggagc ccgtatttgg aaagatgttc gagctgacct
     4981 gcactctgcc tctggagaag gacctaaaga tcactctcta tgactatgac ctcctctcca
     5041 aggacgaaaa gatcggtgag acggtcgtcg acctggagaa caggctgctg tccaagtttg
     5101 gggctcgctg tggactccca cagacctact gtgtctctgg accgaaccag tggcgggacc
     5161 agctccgccc ctcccagctc ctccacctct tctgccagca gcatagagtc aaggcacctg
     5221 tgtaccggac agaccgtgta atgtttcagg ataaagaata ttccattgaa gagatagagg
     5281 ctggcaggat cccaaaccca cacctgggcc cagtggagga gcgtctggct ctgcatgtgc
     5341 ttcagcagca gggcctggtc ccggagcacg tggagtcacg gcccctctac agccccctgc
     5401 agccagacat cgagcagggg aagctgcaga tgtgggtcga cctatttccg aaggccctgg
     5461 ggcggcctgg acctcccttc aacatcaccc cacggagagc cagaaggttt ttcctgcgtt
     5521 gtattatctg gaataccaga gatgtgatcc tggatgacct gagcctcacg ggggagaaga
     5581 tgagcgacat ttatgtgaaa ggttggatga ttggctttga agaacacaag caaaagacag
     5641 acgtgcatta tcgttccctg ggaggtgaag gcaacttcaa ctggaggttc attttcccct
     5701 tcgactacct gccagctgag caagtctgta ccattgccaa gaaggatgcc ttctggaggc
     5761 tggacaagac tgagagcaaa atcccagcac gagtggtgtt ccagatctgg gacaatgaca
     5821 agttctcctt tgatgatttt ctgggctccc tgcagctcga tctcaaccgc atgcccaagc
     5881 cagccaagac agccaagaag tgctccttgg accagctgga tgatgctttc cacccagaat
     5941 ggtttgtgtc cctttttgag cagaaaacag tgaagggctg gtggccctgt gtagcagaag
     6001 agggtgagaa gaaaatactg gcgggcaagc tggaaatgac cttggagatt gtagcagaga
     6061 gtgagcatga ggagcggcct gctggccagg gccgggatga gcccaacatg aaccctaagc
     6121 ttgaggaccc aaggcgcccc gacacctcct tcctgtggtt tacctcccca tacaagacca
     6181 tgaagttcat cctgtggcgg cgtttccggt gggccatcat cctcttcatc atcctcttca
     6241 tcctgctgct gttcctggcc atcttcatct acgccttccc gaactatgct gccatgaagc
     6301 tggtgaagcc cttcagctga tgatcaccat ggcatatggc tagcgcggcc gccatggtca
     6361 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6421 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6481 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6541 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6601 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6661 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6721 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6781 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6841 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6901 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6961 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7021 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7081 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7141 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7201 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7261 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7321 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7381 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7441 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7501 taaacttggt ctgacagtta gaaaaactca tcgagcatca aatgaaactg caatttattc
     7561 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     7621 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     7681 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     7741 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     7801 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     7861 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     7921 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     7981 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccagg gatcgcagtg
     8041 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     8101 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     8161 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     8221 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     8281 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc
     8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8461 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8521 aggccctttc gtc
//
