LOCUS       sequence-96276-e        8518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 43 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8518)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8518)
  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            "Unpublished"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-Exon43-del
FEATURES             Location/Qualifiers
     source          1..8518
                     /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      6333..6342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6342..6358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6342..6358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6355..6377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6366..6382
                     /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(6390..6396,6397..6414,6415..6420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6435..6456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6573..6590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6744..7332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6824..6843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7503..8318)
                     /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(7503..8318)
                     /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     8226..8245
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8319..8423)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8491..8509
                     /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 tttaagtgtg
     4861 atccttacat caagatctcc atagggaaga aatcagtgag tgaccaggat aactacatcc
     4921 cctgcacgct ggagcccgta tttggaaaga tgttcgagct gacctgcact ctgcctctgg
     4981 agaaggacct aaagatcact ctctatgact atgacctcct ctccaaggac gaaaagatcg
     5041 gtgagacggt cgtcgacctg gagaacaggc tgctgtccaa gtttggggct cgctgtggac
     5101 tcccacagac ctactgtgtc tctggaccga accagtggcg ggaccagctc cgcccctccc
     5161 agctcctcca cctcttctgc cagcagcata gagtcaaggc acctgtgtac cggacagacc
     5221 gtgtaatgtt tcaggataaa gaatattcca ttgaagagat agaggctggc aggatcccaa
     5281 acccacacct gggcccagtg gaggagcgtc tggctctgca tgtgcttcag cagcagggcc
     5341 tggtcccgga gcacgtggag tcacggcccc tctacagccc cctgcagcca gacatcgagc
     5401 aggggaagct gcagatgtgg gtcgacctat ttccgaaggc cctggggcgg cctggacctc
     5461 ccttcaacat caccccacgg agagccagaa ggtttttcct gcgttgtatt atctggaata
     5521 ccagagatgt gatcctggat gacctgagcc tcacggggga gaagatgagc gacatttatg
     5581 tgaaaggttg gatgattggc tttgaagaac acaagcaaaa gacagacgtg cattatcgtt
     5641 ccctgggagg tgaaggcaac ttcaactgga ggttcatttt ccccttcgac tacctgccag
     5701 ctgagcaagt ctgtaccatt gccaagaagg atgccttctg gaggctggac aagactgaga
     5761 gcaaaatccc agcacgagtg gtgttccaga tctgggacaa tgacaagttc tcctttgatg
     5821 attttctggg ctccctgcag ctcgatctca accgcatgcc caagccagcc aagacagcca
     5881 agaagtgctc cttggaccag ctggatgatg ctttccaccc agaatggttt gtgtcccttt
     5941 ttgagcagaa aacagtgaag ggctggtggc cctgtgtagc agaagagggt gagaagaaaa
     6001 tactggcggg caagctggaa atgaccttgg agattgtagc agagagtgag catgaggagc
     6061 ggcctgctgg ccagggccgg gatgagccca acatgaaccc taagcttgag gacccaaggc
     6121 gccccgacac ctccttcctg tggtttacct ccccatacaa gaccatgaag ttcatcctgt
     6181 ggcggcgttt ccggtgggcc atcatcctct tcatcatcct cttcatcctg ctgctgttcc
     6241 tggccatctt catctacgcc ttcccgaact atgctgccat gaagctggtg aagcccttca
     6301 gctgatgatc accatggcat atggctagcg cggccgccat ggtcatagct gtttcctgtg
     6361 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6421 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6481 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6541 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6601 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6661 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6721 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6781 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6841 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6901 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6961 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7021 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7081 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7141 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7201 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7261 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7321 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7381 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7441 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7501 agttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     7561 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     7621 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     7681 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     7741 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     7801 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     7861 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     7921 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     7981 tattcttcta atacctggaa tgctgttttc ccagggatcg cagtggtgag taaccatgca
     8041 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     8101 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     8161 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     8221 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     8281 ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     8341 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8401 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8461 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
