LOCUS       sequence-96289-e        8545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 42 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8545)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8545)
  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            "Unpublished"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-Exon42-del
FEATURES             Location/Qualifiers
     source          1..8545
                     /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      6360..6369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6369..6385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6369..6385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6382..6404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6393..6409
                     /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(6417..6423,6424..6441,6442..6447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6462..6483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6600..6617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6771..7359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6851..6870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7530..8345)
                     /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(7530..8345)
                     /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     8253..8272
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8346..8450)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8518..8536
                     /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 cctgaagggc ctcttcaaaa
     4741 tttatcccct cccagaagac ccagccatcc ccatgccccc aagacagttc caccagctgg
     4801 ccgcccaggg accccaggag tgcttggtcc gtatctacat tgtccgagca tttggcctgc
     4861 agcccaagga ccccaatgga aagtgtgatc cttacatcaa gatctccata gggaagaaat
     4921 cagtgagtga ccaggataac tacatcccct gcacgctgga gcccgtattt ggaaagatgt
     4981 tcgagctgac ctgcactctg cctctggaga aggacctaaa gatcactctc tatgactatg
     5041 acctcctctc caaggacgaa aagatcggtg agacggtcgt cgacctggag aacaggctgc
     5101 tgtccaagtt tggggctcgc tgtggactcc cacagaccta ctgtgtctct ggaccgaacc
     5161 agtggcggga ccagctccgc ccctcccagc tcctccacct cttctgccag cagcatagag
     5221 tcaaggcacc tgtgtaccgg acagaccgtg taatgtttca ggataaagaa tattccattg
     5281 aagagataga ggctggcagg atcccaaacc cacacctggg cccagtggag gagcgtctgg
     5341 ctctgcatgt gcttcagcag cagggcctgg tcccggagca cgtggagtca cggcccctct
     5401 acagccccct gcagccagac atcgagcagg ggaagctgca gatgtgggtc gacctatttc
     5461 cgaaggccct ggggcggcct ggacctccct tcaacatcac cccacggaga gccagaaggt
     5521 ttttcctgcg ttgtattatc tggaatacca gagatgtgat cctggatgac ctgagcctca
     5581 cgggggagaa gatgagcgac atttatgtga aaggttggat gattggcttt gaagaacaca
     5641 agcaaaagac agacgtgcat tatcgttccc tgggaggtga aggcaacttc aactggaggt
     5701 tcattttccc cttcgactac ctgccagctg agcaagtctg taccattgcc aagaaggatg
     5761 ccttctggag gctggacaag actgagagca aaatcccagc acgagtggtg ttccagatct
     5821 gggacaatga caagttctcc tttgatgatt ttctgggctc cctgcagctc gatctcaacc
     5881 gcatgcccaa gccagccaag acagccaaga agtgctcctt ggaccagctg gatgatgctt
     5941 tccacccaga atggtttgtg tccctttttg agcagaaaac agtgaagggc tggtggccct
     6001 gtgtagcaga agagggtgag aagaaaatac tggcgggcaa gctggaaatg accttggaga
     6061 ttgtagcaga gagtgagcat gaggagcggc ctgctggcca gggccgggat gagcccaaca
     6121 tgaaccctaa gcttgaggac ccaaggcgcc ccgacacctc cttcctgtgg tttacctccc
     6181 catacaagac catgaagttc atcctgtggc ggcgtttccg gtgggccatc atcctcttca
     6241 tcatcctctt catcctgctg ctgttcctgg ccatcttcat ctacgccttc ccgaactatg
     6301 ctgccatgaa gctggtgaag cccttcagct gatgatcacc atggcatatg gctagcgcgg
     6361 ccgccatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6421 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6481 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6541 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6601 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6661 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6721 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6781 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6961 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7141 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7201 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7261 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7321 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7381 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7441 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7501 agtatatatg agtaaacttg gtctgacagt tagaaaaact catcgagcat caaatgaaac
     7561 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     7621 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     7681 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     7741 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     7801 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     7861 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     7921 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     7981 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccca
     8041 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     8101 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     8161 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     8221 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     8281 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     8341 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8401 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8461 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     8521 aggcgtatca cgaggccctt tcgtc
//
