LOCUS       sequence-96267-e        8545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 7 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  JOURNAL   Unpublished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            "Unpublished"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-Exon7-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"
     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 gactcttttc ttcaacttgt
      901 ttgactctcc tggggagctg tttgatgagc ccatctttat cacggtggta gactctcgtt
      961 ctctcaggac agatgctctc ctcggggagt tccggatgga cgtgggcacc atttacagag
     1021 agccccggca cgcctatctc aggaagtggc tgctgctctc agaccctgat gacttctctg
     1081 ctggggccag aggctacctg aaaacaagcc tttgtgtgct ggggcctggg gacgaagcgc
     1141 ctctggagag aaaagacccc tctgaagaca aggaggacat tgaaagcaac ctgctccggc
     1201 ccacaggcgt agccctgcga ggagcccact tctgcctgaa ggtcttccgg gccgaggact
     1261 tgccgcagat ggacgatgcc gtgatggaca acgtgaaaca gatctttggc ttcgagagta
     1321 acaagaagaa cttggtggac ccctttgtgg aggtcagctt tgcggggaaa atgctgtgca
     1381 gcaagatctt ggagaagacg gccaaccctc agtggaacca gaacatcaca ctgcctgcca
     1441 tgtttccctc catgtgcgaa aaaatgagga ttcgtatcat agactgggac cgcctgactc
     1501 acaatgacat cgtggctacc acctacctga gtatgtcgaa aatctctgcc cctggaggag
     1561 aaatagaaga ggagcctgca ggtgctgtca agccttcgaa agcctcagac ttggatgact
     1621 acctgggctt cctccccact tttgggccct gctacatcaa cctctatggc agtcccagag
     1681 agttcacagg cttcccagac ccctacacag agctcaacac aggcaagggg gaaggtgtgg
     1741 cttatcgtgg ccggcttctg ctctccctgg agaccaagct ggtggagcac agtgaacaga
     1801 aggtggagga ccttcctgcg gatgacatcc tccgggtgga gaagtacctt aggaggcgca
     1861 agtactccct gtttgcggcc ttctactcag ccaccatgct gcaggatgtg gatgatgcca
     1921 tccagtttga ggtcagcatc gggaactacg ggaacaagtt cgacatgacc tgcctgccgc
     1981 tggcctccac cactcagtac agccgtgcag tctttgacgg gtgccactac tactacctac
     2041 cctggggtaa cgtgaaacct gtggtggtgc tgtcatccta ctgggaggac atcagccata
     2101 gaatcgagac tcagaaccag ctgcttggga ttgctgaccg gctggaagct ggcctggagc
     2161 aggtccacct ggccctgaag gcgcagtgct ccacggagga cgtggactcg ctggtggctc
     2221 agctgacgga tgagctcatc gcaggctgca gccagcctct gggtgacatc catgagacac
     2281 cctctgccac ccacctggac cagtacctgt accagctgcg cacccatcac ctgagccaaa
     2341 tcactgaggc tgccctggcc ctgaagctcg gccacagtga gctccctgca gctctggagc
     2401 aggcggagga ctggctcctg cgtctgcgtg ccctggcaga ggagccccag aacagcctgc
     2461 cggacatcgt catctggatg ctgcagggag acaagcgtgt ggcataccag cgggtgcccg
     2521 cccaccaagt cctcttctcc cggcggggtg ccaactactg tggcaagaat tgtgggaagc
     2581 tacagacaat ctttctgaaa tatccgatgg agaaggtgcc tggcgcccgg atgccagtgc
     2641 agatacgggt caagctgtgg tttgggctct ctgtggatga gaaggagttc aaccagtttg
     2701 ctgaggggaa gctgtctgtc tttgctgaaa cctatgagaa cgagactaag ttggcccttg
     2761 ttgggaactg gggcacaacg ggcctcacct accccaagtt ttctgacgtc acgggcaaga
     2821 tcaagctacc caaggacagc ttccgcccct cggccggctg gacctgggct ggagattggt
     2881 tcgtgtgtcc ggagaagact ctgctccatg acatggacgc cggtcacctg agcttcgtgg
     2941 aagaggtgtt tgagaaccag acccggcttc ccggaggcca gtggatctac atgagtgaca
     3001 actacaccga tgtgaacggg gagaaggtgc ttcccaagga tgacattgag tgcccactgg
     3061 gctggaagtg ggaagatgag gaatggtcca cagacctcaa ccgggctgtc gatgagcaag
     3121 gctgggagta tagcatcacc atccccccgg agcggaagcc gaagcactgg gtccctgctg
     3181 agaagatgta ctacacacac cgacggcggc gctgggtgcg cctgcgcagg agggatctca
     3241 gccaaatgga agcactgaaa aggcacaggc aggcggaggc ggagggcgag ggctgggagt
     3301 acgcctctct ttttggctgg aagttccacc tcgagtaccg caagacagat gccttccgcc
     3361 gccgccgctg gcgccgtcgc atggagccac tggagaagac ggggcctgca gctgtgtttg
     3421 cccttgaggg ggccctgggc ggcgtgatgg atgacaagag tgaagattcc atgtccgtct
     3481 ccaccttgag cttcggtgtg aacagaccca cgatttcctg catattcgac tatgggaacc
     3541 gctaccatct acgctgctac atgtaccagg cccgggacct ggctgcgatg gacaaggact
     3601 ctttttctga tccctatgcc atcgtctcct tcctgcacca gagccagaag acggtggtgg
     3661 tgaagaacac ccttaacccc acctgggacc agacgctcat cttctacgag atcgagatct
     3721 ttggcgagcc ggccacagtt gctgagcaac cgcccagcat tgtggtggag ctgtacgacc
     3781 atgacactta tggtgcagac gagtttatgg gtcgctgcat ctgtcaaccg agtctggaac
     3841 ggatgccacg gctggcctgg ttcccactga cgaggggcag ccagccgtcg ggggagctgc
     3901 tggcctcttt tgagctcatc cagagagaga agccggccat ccaccatatt cctggttttg
     3961 aggtgcagga gacatcaagg atcctggatg agtctgagga cacagacctg ccctacccac
     4021 caccccagag ggaggccaac atctacatgg ttcctcagaa catcaagcca gcgctccagc
     4081 gtaccgccat cgagatcctg gcatggggcc tgcggaacat gaagagttac cagctggcca
     4141 acatctcctc ccccagcctc gtggtagagt gtgggggcca gacggtgcag tcctgtgtca
     4201 tcaggaacct ccggaagaac cccaactttg acatctgcac cctcttcatg gaagtgatgc
     4261 tgcccaggga ggagctctac tgccccccca tcaccgtcaa ggtcatcgat aaccgccagt
     4321 ttggccgccg gcctgtggtg ggccagtgta ccatccgctc cctggagagc ttcctgtgtg
     4381 acccctactc ggcggagagt ccatccccac agggtggccc agacgatgtg agcctactca
     4441 gtcctgggga agacgtgctc atcgacattg atgacaagga gcccctcatc cccatccagg
     4501 aggaagagtt catcgattgg tggagcaaat tctttgcctc cataggggag agggaaaagt
     4561 gcggctccta cctggagaag gattttgaca ccctgaaggt ctatgacaca cagctggaga
     4621 atgtggaggc ctttgagggc ctgtctgact tttgtaacac cttcaagctg taccggggca
     4681 agacgcagga ggagacagaa gatccatctg tgattggtga atttaagggc 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
//
