LOCUS       sequence-96288-e        8575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 41 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8575)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8575)
  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            "Unpublished"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-Exon41-del
FEATURES             Location/Qualifiers
     source          1..8575
                     /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      6390..6399
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6399..6415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6399..6415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6412..6434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6423..6439
                     /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(6447..6453,6454..6471,6472..6477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6492..6513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6630..6647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6801..7389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6881..6900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7560..8375)
                     /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(7560..8375)
                     /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     8283..8302
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8376..8480)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8548..8566
                     /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 ggtgcaggag acatcaagga tcctggatga gtctgaggac acagacctgc
     4141 cctacccacc accccagagg gaggccaaca tctacatggt tcctcagaac atcaagccag
     4201 cgctccagcg taccgccatc gagatcctgg catggggcct gcggaacatg aagagttacc
     4261 agctggccaa catctcctcc cccagcctcg tggtagagtg tgggggccag acggtgcagt
     4321 cctgtgtcat caggaacctc cggaagaacc ccaactttga catctgcacc ctcttcatgg
     4381 aagtgatgct gcccagggag gagctctact gcccccccat caccgtcaag gtcatcgata
     4441 accgccagtt tggccgccgg cctgtggtgg gccagtgtac catccgctcc ctggagagct
     4501 tcctgtgtga cccctactcg gcggagagtc catccccaca gggtggccca gacgatgtga
     4561 gcctactcag tcctggggaa gacgtgctca tcgacattga tgacaaggag cccctcatcc
     4621 ccatccaggt ctatgacaca cagctggaga atgtggaggc ctttgagggc ctgtctgact
     4681 tttgtaacac cttcaagctg taccggggca agacgcagga ggagacagaa gatccatctg
     4741 tgattggtga atttaagggc ctcttcaaaa tttatcccct cccagaagac ccagccatcc
     4801 ccatgccccc aagacagttc caccagctgg ccgcccaggg accccaggag tgcttggtcc
     4861 gtatctacat tgtccgagca tttggcctgc agcccaagga ccccaatgga aagtgtgatc
     4921 cttacatcaa gatctccata gggaagaaat cagtgagtga ccaggataac tacatcccct
     4981 gcacgctgga gcccgtattt ggaaagatgt tcgagctgac ctgcactctg cctctggaga
     5041 aggacctaaa gatcactctc tatgactatg acctcctctc caaggacgaa aagatcggtg
     5101 agacggtcgt cgacctggag aacaggctgc tgtccaagtt tggggctcgc tgtggactcc
     5161 cacagaccta ctgtgtctct ggaccgaacc agtggcggga ccagctccgc ccctcccagc
     5221 tcctccacct cttctgccag cagcatagag tcaaggcacc tgtgtaccgg acagaccgtg
     5281 taatgtttca ggataaagaa tattccattg aagagataga ggctggcagg atcccaaacc
     5341 cacacctggg cccagtggag gagcgtctgg ctctgcatgt gcttcagcag cagggcctgg
     5401 tcccggagca cgtggagtca cggcccctct acagccccct gcagccagac atcgagcagg
     5461 ggaagctgca gatgtgggtc gacctatttc cgaaggccct ggggcggcct ggacctccct
     5521 tcaacatcac cccacggaga gccagaaggt ttttcctgcg ttgtattatc tggaatacca
     5581 gagatgtgat cctggatgac ctgagcctca cgggggagaa gatgagcgac atttatgtga
     5641 aaggttggat gattggcttt gaagaacaca agcaaaagac agacgtgcat tatcgttccc
     5701 tgggaggtga aggcaacttc aactggaggt tcattttccc cttcgactac ctgccagctg
     5761 agcaagtctg taccattgcc aagaaggatg ccttctggag gctggacaag actgagagca
     5821 aaatcccagc acgagtggtg ttccagatct gggacaatga caagttctcc tttgatgatt
     5881 ttctgggctc cctgcagctc gatctcaacc gcatgcccaa gccagccaag acagccaaga
     5941 agtgctcctt ggaccagctg gatgatgctt tccacccaga atggtttgtg tccctttttg
     6001 agcagaaaac agtgaagggc tggtggccct gtgtagcaga agagggtgag aagaaaatac
     6061 tggcgggcaa gctggaaatg accttggaga ttgtagcaga gagtgagcat gaggagcggc
     6121 ctgctggcca gggccgggat gagcccaaca tgaaccctaa gcttgaggac ccaaggcgcc
     6181 ccgacacctc cttcctgtgg tttacctccc catacaagac catgaagttc atcctgtggc
     6241 ggcgtttccg gtgggccatc atcctcttca tcatcctctt catcctgctg ctgttcctgg
     6301 ccatcttcat ctacgccttc ccgaactatg ctgccatgaa gctggtgaag cccttcagct
     6361 gatgatcacc atggcatatg gctagcgcgg ccgccatggt catagctgtt tcctgtgtga
     6421 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6481 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6541 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6601 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6661 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7201 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7381 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7501 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7561 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     7621 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     7681 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     7741 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     7801 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     7861 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     7921 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     7981 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     8041 tcttctaata cctggaatgc tgttttccca gggatcgcag tggtgagtaa ccatgcatca
     8101 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     8161 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     8221 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     8281 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     8341 ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga
     8401 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8461 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     8521 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
