LOCUS       sequence-96290-e        8503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 45 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8503)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8503)
  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            "Unpublished"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-Exon45-del
FEATURES             Location/Qualifiers
     source          1..8503
                     /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      6318..6327
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6327..6343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6327..6343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6340..6362)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6351..6367
                     /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(6375..6381,6382..6399,6400..6405))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6420..6441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6558..6575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6729..7317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6809..6828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7488..8303)
                     /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(7488..8303)
                     /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     8211..8230
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8304..8408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8476..8494
                     /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 ccatccagga ggaagagttc atcgattggt ggagcaaatt ctttgcctcc ataggggaga
     4681 gggaaaagtg cggctcctac ctggagaagg attttgacac cctgaaggtc tatgacacac
     4741 agctggagaa tgtggaggcc tttgagggcc tgtctgactt ttgtaacacc ttcaagctgt
     4801 accggggcaa gacgcaggag gagacagaag atccatctgt gattggtgaa tttaagggcc
     4861 tcttcaaaat ttatcccctc ccagaagacc cagccatccc catgccccca agacagttcc
     4921 accagctggc cgcccaggga ccccaggagt gcttggtccg tatctacatt gtccgagcat
     4981 ttggcctgca gcccaaggac cccaatggaa agtgtgatcc ttacatcaag atctccatag
     5041 ggaagaaatc agtgagtgac caggataact acatcccctg cacgctggag cccgtatttg
     5101 gaaactctgg accgaaccag tggcgggacc agctccgccc ctcccagctc ctccacctct
     5161 tctgccagca gcatagagtc aaggcacctg tgtaccggac agaccgtgta atgtttcagg
     5221 ataaagaata ttccattgaa gagatagagg ctggcaggat cccaaaccca cacctgggcc
     5281 cagtggagga gcgtctggct ctgcatgtgc ttcagcagca gggcctggtc ccggagcacg
     5341 tggagtcacg gcccctctac agccccctgc agccagacat cgagcagggg aagctgcaga
     5401 tgtgggtcga cctatttccg aaggccctgg ggcggcctgg acctcccttc aacatcaccc
     5461 cacggagagc cagaaggttt ttcctgcgtt gtattatctg gaataccaga gatgtgatcc
     5521 tggatgacct gagcctcacg ggggagaaga tgagcgacat ttatgtgaaa ggttggatga
     5581 ttggctttga agaacacaag caaaagacag acgtgcatta tcgttccctg ggaggtgaag
     5641 gcaacttcaa ctggaggttc attttcccct tcgactacct gccagctgag caagtctgta
     5701 ccattgccaa gaaggatgcc ttctggaggc tggacaagac tgagagcaaa atcccagcac
     5761 gagtggtgtt ccagatctgg gacaatgaca agttctcctt tgatgatttt ctgggctccc
     5821 tgcagctcga tctcaaccgc atgcccaagc cagccaagac agccaagaag tgctccttgg
     5881 accagctgga tgatgctttc cacccagaat ggtttgtgtc cctttttgag cagaaaacag
     5941 tgaagggctg gtggccctgt gtagcagaag agggtgagaa gaaaatactg gcgggcaagc
     6001 tggaaatgac cttggagatt gtagcagaga gtgagcatga ggagcggcct gctggccagg
     6061 gccgggatga gcccaacatg aaccctaagc ttgaggaccc aaggcgcccc gacacctcct
     6121 tcctgtggtt tacctcccca tacaagacca tgaagttcat cctgtggcgg cgtttccggt
     6181 gggccatcat cctcttcatc atcctcttca tcctgctgct gttcctggcc atcttcatct
     6241 acgccttccc gaactatgct gccatgaagc tggtgaagcc cttcagctga tgatcaccat
     6301 ggcatatggc tagcgcggcc gccatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6361 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6421 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6481 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6541 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6601 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6661 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6721 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6781 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6841 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6901 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6961 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7021 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7081 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7141 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7201 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7261 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7321 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7381 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7441 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca
     7501 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     7561 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     7621 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     7681 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     7741 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     7801 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     7861 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     7921 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     7981 tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     8041 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     8101 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     8161 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     8221 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     8281 gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag catttatcag
     8341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8401 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8461 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
