LOCUS       sequence-96285-e        8644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 36 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8644)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8644)
  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            "Unpublished"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-Exon36-del
FEATURES             Location/Qualifiers
     source          1..8644
                     /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      6459..6468
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6468..6484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6468..6484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6481..6503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6492..6508
                     /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(6516..6522,6523..6540,6541..6546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6561..6582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6699..6716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6870..7458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6950..6969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7629..8444)
                     /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(7629..8444)
                     /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     8352..8371
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8445..8549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8617..8635
                     /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 ggtgcagacg agtttatggg tcgctgcatc tgtcaaccga
     3961 gtctggaacg gatgccacgg ctggcctggt tcccactgac gaggggcagc cagccgtcgg
     4021 gggagctgct ggcctctttt gagctcatcc agagagagaa gccggccatc caccatattc
     4081 ctggttttga gtctgaggac acagacctgc cctacccacc accccagagg gaggccaaca
     4141 tctacatggt tcctcagaac atcaagccag cgctccagcg taccgccatc gagatcctgg
     4201 catggggcct gcggaacatg aagagttacc agctggccaa catctcctcc cccagcctcg
     4261 tggtagagtg tgggggccag acggtgcagt cctgtgtcat caggaacctc cggaagaacc
     4321 ccaactttga catctgcacc ctcttcatgg aagtgatgct gcccagggag gagctctact
     4381 gcccccccat caccgtcaag gtcatcgata accgccagtt tggccgccgg cctgtggtgg
     4441 gccagtgtac catccgctcc ctggagagct tcctgtgtga cccctactcg gcggagagtc
     4501 catccccaca gggtggccca gacgatgtga gcctactcag tcctggggaa gacgtgctca
     4561 tcgacattga tgacaaggag cccctcatcc ccatccagga ggaagagttc atcgattggt
     4621 ggagcaaatt ctttgcctcc ataggggaga gggaaaagtg cggctcctac ctggagaagg
     4681 attttgacac cctgaaggtc tatgacacac agctggagaa tgtggaggcc tttgagggcc
     4741 tgtctgactt ttgtaacacc ttcaagctgt accggggcaa gacgcaggag gagacagaag
     4801 atccatctgt gattggtgaa tttaagggcc tcttcaaaat ttatcccctc ccagaagacc
     4861 cagccatccc catgccccca agacagttcc accagctggc cgcccaggga ccccaggagt
     4921 gcttggtccg tatctacatt gtccgagcat ttggcctgca gcccaaggac cccaatggaa
     4981 agtgtgatcc ttacatcaag atctccatag ggaagaaatc agtgagtgac caggataact
     5041 acatcccctg cacgctggag cccgtatttg gaaagatgtt cgagctgacc tgcactctgc
     5101 ctctggagaa ggacctaaag atcactctct atgactatga cctcctctcc aaggacgaaa
     5161 agatcggtga gacggtcgtc gacctggaga acaggctgct gtccaagttt ggggctcgct
     5221 gtggactccc acagacctac tgtgtctctg gaccgaacca gtggcgggac cagctccgcc
     5281 cctcccagct cctccacctc ttctgccagc agcatagagt caaggcacct gtgtaccgga
     5341 cagaccgtgt aatgtttcag gataaagaat attccattga agagatagag gctggcagga
     5401 tcccaaaccc acacctgggc ccagtggagg agcgtctggc tctgcatgtg cttcagcagc
     5461 agggcctggt cccggagcac gtggagtcac ggcccctcta cagccccctg cagccagaca
     5521 tcgagcaggg gaagctgcag atgtgggtcg acctatttcc gaaggccctg gggcggcctg
     5581 gacctccctt caacatcacc ccacggagag ccagaaggtt tttcctgcgt tgtattatct
     5641 ggaataccag agatgtgatc ctggatgacc tgagcctcac gggggagaag atgagcgaca
     5701 tttatgtgaa aggttggatg attggctttg aagaacacaa gcaaaagaca gacgtgcatt
     5761 atcgttccct gggaggtgaa ggcaacttca actggaggtt cattttcccc ttcgactacc
     5821 tgccagctga gcaagtctgt accattgcca agaaggatgc cttctggagg ctggacaaga
     5881 ctgagagcaa aatcccagca cgagtggtgt tccagatctg ggacaatgac aagttctcct
     5941 ttgatgattt tctgggctcc ctgcagctcg atctcaaccg catgcccaag ccagccaaga
     6001 cagccaagaa gtgctccttg gaccagctgg atgatgcttt ccacccagaa tggtttgtgt
     6061 ccctttttga gcagaaaaca gtgaagggct ggtggccctg tgtagcagaa gagggtgaga
     6121 agaaaatact ggcgggcaag ctggaaatga ccttggagat tgtagcagag agtgagcatg
     6181 aggagcggcc tgctggccag ggccgggatg agcccaacat gaaccctaag cttgaggacc
     6241 caaggcgccc cgacacctcc ttcctgtggt ttacctcccc atacaagacc atgaagttca
     6301 tcctgtggcg gcgtttccgg tgggccatca tcctcttcat catcctcttc atcctgctgc
     6361 tgttcctggc catcttcatc tacgccttcc cgaactatgc tgccatgaag ctggtgaagc
     6421 ccttcagctg atgatcacca tggcatatgg ctagcgcggc cgccatggtc atagctgttt
     6481 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6541 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6601 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6661 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7621 tctgacagtt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     7681 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     7741 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     7801 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     7861 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     7921 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     7981 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     8041 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     8101 tcaggatatt cttctaatac ctggaatgct gttttcccag ggatcgcagt ggtgagtaac
     8161 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     8221 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     8281 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     8341 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     8401 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa
     8461 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8521 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8581 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     8641 cgtc
//
