LOCUS       sequence-96282-e        8596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 32 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8596)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8596)
  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            "Unpublished"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-Exon32-del
FEATURES             Location/Qualifiers
     source          1..8596
                     /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      6411..6420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6420..6436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6420..6436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6433..6455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6444..6460
                     /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(6468..6474,6475..6492,6493..6498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6513..6534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6651..6668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6822..7410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6902..6921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7581..8396)
                     /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(7581..8396)
                     /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     8304..8323
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8397..8501)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8569..8587
                     /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 atccctatgc catcgtctcc ttcctgcacc agagccagaa gacggtggtg gtgaagaaca
     3721 cccttaaccc cacctgggac cagacgctca tcttctacga gatcgagatc tttggcgagc
     3781 cggccacagt tgctgagcaa ccgcccagca ttgtggtgga gctgtacgac catgacactt
     3841 atggtgcaga cgagtttatg ggtcgctgca tctgtcaacc gagtctggaa cggatgccac
     3901 ggctggcctg gttcccactg acgaggggca gccagccgtc gggggagctg ctggcctctt
     3961 ttgagctcat ccagagagag aagccggcca tccaccatat tcctggtttt gaggtgcagg
     4021 agacatcaag gatcctggat gagtctgagg acacagacct gccctaccca ccaccccaga
     4081 gggaggccaa catctacatg gttcctcaga acatcaagcc agcgctccag cgtaccgcca
     4141 tcgagatcct ggcatggggc ctgcggaaca tgaagagtta ccagctggcc aacatctcct
     4201 cccccagcct cgtggtagag tgtgggggcc agacggtgca gtcctgtgtc atcaggaacc
     4261 tccggaagaa ccccaacttt gacatctgca ccctcttcat ggaagtgatg ctgcccaggg
     4321 aggagctcta ctgccccccc atcaccgtca aggtcatcga taaccgccag tttggccgcc
     4381 ggcctgtggt gggccagtgt accatccgct ccctggagag cttcctgtgt gacccctact
     4441 cggcggagag tccatcccca cagggtggcc cagacgatgt gagcctactc agtcctgggg
     4501 aagacgtgct catcgacatt gatgacaagg agcccctcat ccccatccag gaggaagagt
     4561 tcatcgattg gtggagcaaa ttctttgcct ccatagggga gagggaaaag tgcggctcct
     4621 acctggagaa ggattttgac accctgaagg tctatgacac acagctggag aatgtggagg
     4681 cctttgaggg cctgtctgac ttttgtaaca ccttcaagct gtaccggggc aagacgcagg
     4741 aggagacaga agatccatct gtgattggtg aatttaaggg cctcttcaaa atttatcccc
     4801 tcccagaaga cccagccatc cccatgcccc caagacagtt ccaccagctg gccgcccagg
     4861 gaccccagga gtgcttggtc cgtatctaca ttgtccgagc atttggcctg cagcccaagg
     4921 accccaatgg aaagtgtgat ccttacatca agatctccat agggaagaaa tcagtgagtg
     4981 accaggataa ctacatcccc tgcacgctgg agcccgtatt tggaaagatg ttcgagctga
     5041 cctgcactct gcctctggag aaggacctaa agatcactct ctatgactat gacctcctct
     5101 ccaaggacga aaagatcggt gagacggtcg tcgacctgga gaacaggctg ctgtccaagt
     5161 ttggggctcg ctgtggactc ccacagacct actgtgtctc tggaccgaac cagtggcggg
     5221 accagctccg cccctcccag ctcctccacc tcttctgcca gcagcataga gtcaaggcac
     5281 ctgtgtaccg gacagaccgt gtaatgtttc aggataaaga atattccatt gaagagatag
     5341 aggctggcag gatcccaaac ccacacctgg gcccagtgga ggagcgtctg gctctgcatg
     5401 tgcttcagca gcagggcctg gtcccggagc acgtggagtc acggcccctc tacagccccc
     5461 tgcagccaga catcgagcag gggaagctgc agatgtgggt cgacctattt ccgaaggccc
     5521 tggggcggcc tggacctccc ttcaacatca ccccacggag agccagaagg tttttcctgc
     5581 gttgtattat ctggaatacc agagatgtga tcctggatga cctgagcctc acgggggaga
     5641 agatgagcga catttatgtg aaaggttgga tgattggctt tgaagaacac aagcaaaaga
     5701 cagacgtgca ttatcgttcc ctgggaggtg aaggcaactt caactggagg ttcattttcc
     5761 ccttcgacta cctgccagct gagcaagtct gtaccattgc caagaaggat gccttctgga
     5821 ggctggacaa gactgagagc aaaatcccag cacgagtggt gttccagatc tgggacaatg
     5881 acaagttctc ctttgatgat tttctgggct ccctgcagct cgatctcaac cgcatgccca
     5941 agccagccaa gacagccaag aagtgctcct tggaccagct ggatgatgct ttccacccag
     6001 aatggtttgt gtcccttttt gagcagaaaa cagtgaaggg ctggtggccc tgtgtagcag
     6061 aagagggtga gaagaaaata ctggcgggca agctggaaat gaccttggag attgtagcag
     6121 agagtgagca tgaggagcgg cctgctggcc agggccggga tgagcccaac atgaacccta
     6181 agcttgagga cccaaggcgc cccgacacct ccttcctgtg gtttacctcc ccatacaaga
     6241 ccatgaagtt catcctgtgg cggcgtttcc ggtgggccat catcctcttc atcatcctct
     6301 tcatcctgct gctgttcctg gccatcttca tctacgcctt cccgaactat gctgccatga
     6361 agctggtgaa gcccttcagc tgatgatcac catggcatat ggctagcgcg gccgccatgg
     6421 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6481 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6541 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6601 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6661 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6721 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6781 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6841 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6901 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6961 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7021 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7081 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7141 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7201 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7261 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7321 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7381 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7441 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7501 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7561 gagtaaactt ggtctgacag ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     7621 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     7681 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     7741 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     7801 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     7861 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     7921 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     7981 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     8041 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc agggatcgca
     8101 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     8161 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     8221 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     8281 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     8341 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc
     8401 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8461 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8521 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     8581 acgaggccct ttcgtc
//
