LOCUS       sequence-96272-e        8623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 9 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8623)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8623)
  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            "Unpublished"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-Exon9-del
FEATURES             Location/Qualifiers
     source          1..8623
                     /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      6438..6447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6447..6463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6447..6463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6460..6482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6471..6487
                     /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(6495..6501,6502..6519,6520..6525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6540..6561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6678..6695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6849..7437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6929..6948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7608..8423)
                     /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(7608..8423)
                     /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     8331..8350
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8424..8528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8596..8614
                     /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 acgatggacg
     1081 tgggcaccat ttacagagag ccccggcacg cctatctcag gaagtggctg ctgctctcag
     1141 accctgatga cttctctgct ggggccagag gctacctgaa aacaagcctt tgtgtgctgg
     1201 ggcctgggga cgaagcgcct ctggagagaa aagacccctc tgaagacaag gaggacattg
     1261 aaagcaacct gctccggccc acaggcgtag ccctgcgagg agcccacttc tgcctgaagg
     1321 tcttccgggc cgaggacttg ccgcagatgg acgatgccgt gatggacaac gtgaaacaga
     1381 tctttggctt cgagagtaac aagaagaact tggtggaccc ctttgtggag gtcagctttg
     1441 cggggaaaat gctgtgcagc aagatcttgg agaagacggc caaccctcag tggaaccaga
     1501 acatcacact gcctgccatg tttccctcca tgtgcgaaaa aatgaggatt cgtatcatag
     1561 actgggaccg cctgactcac aatgacatcg tggctaccac ctacctgagt atgtcgaaaa
     1621 tctctgcccc tggaggagaa atagaagagg agcctgcagg tgctgtcaag ccttcgaaag
     1681 cctcagactt ggatgactac ctgggcttcc tccccacttt tgggccctgc tacatcaacc
     1741 tctatggcag tcccagagag ttcacaggct tcccagaccc ctacacagag ctcaacacag
     1801 gcaaggggga aggtgtggct tatcgtggcc ggcttctgct ctccctggag accaagctgg
     1861 tggagcacag tgaacagaag gtggaggacc ttcctgcgga tgacatcctc cgggtggaga
     1921 agtaccttag gaggcgcaag tactccctgt ttgcggcctt ctactcagcc accatgctgc
     1981 aggatgtgga tgatgccatc cagtttgagg tcagcatcgg gaactacggg aacaagttcg
     2041 acatgacctg cctgccgctg gcctccacca ctcagtacag ccgtgcagtc tttgacgggt
     2101 gccactacta ctacctaccc tggggtaacg tgaaacctgt ggtggtgctg tcatcctact
     2161 gggaggacat cagccataga atcgagactc agaaccagct gcttgggatt gctgaccggc
     2221 tggaagctgg cctggagcag gtccacctgg ccctgaaggc gcagtgctcc acggaggacg
     2281 tggactcgct ggtggctcag ctgacggatg agctcatcgc aggctgcagc cagcctctgg
     2341 gtgacatcca tgagacaccc tctgccaccc acctggacca gtacctgtac cagctgcgca
     2401 cccatcacct gagccaaatc actgaggctg ccctggccct gaagctcggc cacagtgagc
     2461 tccctgcagc tctggagcag gcggaggact ggctcctgcg tctgcgtgcc ctggcagagg
     2521 agccccagaa cagcctgccg gacatcgtca tctggatgct gcagggagac aagcgtgtgg
     2581 cataccagcg ggtgcccgcc caccaagtcc tcttctcccg gcggggtgcc aactactgtg
     2641 gcaagaattg tgggaagcta cagacaatct ttctgaaata tccgatggag aaggtgcctg
     2701 gcgcccggat gccagtgcag atacgggtca agctgtggtt tgggctctct gtggatgaga
     2761 aggagttcaa ccagtttgct gaggggaagc tgtctgtctt tgctgaaacc tatgagaacg
     2821 agactaagtt ggcccttgtt gggaactggg gcacaacggg cctcacctac cccaagtttt
     2881 ctgacgtcac gggcaagatc aagctaccca aggacagctt ccgcccctcg gccggctgga
     2941 cctgggctgg agattggttc gtgtgtccgg agaagactct gctccatgac atggacgccg
     3001 gtcacctgag cttcgtggaa gaggtgtttg agaaccagac ccggcttccc ggaggccagt
     3061 ggatctacat gagtgacaac tacaccgatg tgaacgggga gaaggtgctt cccaaggatg
     3121 acattgagtg cccactgggc tggaagtggg aagatgagga atggtccaca gacctcaacc
     3181 gggctgtcga tgagcaaggc tgggagtata gcatcaccat ccccccggag cggaagccga
     3241 agcactgggt ccctgctgag aagatgtact acacacaccg acggcggcgc tgggtgcgcc
     3301 tgcgcaggag ggatctcagc caaatggaag cactgaaaag gcacaggcag gcggaggcgg
     3361 agggcgaggg ctgggagtac gcctctcttt ttggctggaa gttccacctc gagtaccgca
     3421 agacagatgc cttccgccgc cgccgctggc gccgtcgcat ggagccactg gagaagacgg
     3481 ggcctgcagc tgtgtttgcc cttgaggggg ccctgggcgg cgtgatggat gacaagagtg
     3541 aagattccat gtccgtctcc accttgagct tcggtgtgaa cagacccacg atttcctgca
     3601 tattcgacta tgggaaccgc taccatctac gctgctacat gtaccaggcc cgggacctgg
     3661 ctgcgatgga caaggactct ttttctgatc cctatgccat cgtctccttc ctgcaccaga
     3721 gccagaagac ggtggtggtg aagaacaccc ttaaccccac ctgggaccag acgctcatct
     3781 tctacgagat cgagatcttt ggcgagccgg ccacagttgc tgagcaaccg cccagcattg
     3841 tggtggagct gtacgaccat gacacttatg gtgcagacga gtttatgggt cgctgcatct
     3901 gtcaaccgag tctggaacgg atgccacggc tggcctggtt cccactgacg aggggcagcc
     3961 agccgtcggg ggagctgctg gcctcttttg agctcatcca gagagagaag ccggccatcc
     4021 accatattcc tggttttgag gtgcaggaga catcaaggat cctggatgag tctgaggaca
     4081 cagacctgcc ctacccacca ccccagaggg aggccaacat ctacatggtt cctcagaaca
     4141 tcaagccagc gctccagcgt accgccatcg agatcctggc atggggcctg cggaacatga
     4201 agagttacca gctggccaac atctcctccc ccagcctcgt ggtagagtgt gggggccaga
     4261 cggtgcagtc ctgtgtcatc aggaacctcc ggaagaaccc caactttgac atctgcaccc
     4321 tcttcatgga agtgatgctg cccagggagg agctctactg cccccccatc accgtcaagg
     4381 tcatcgataa ccgccagttt ggccgccggc ctgtggtggg ccagtgtacc atccgctccc
     4441 tggagagctt cctgtgtgac ccctactcgg cggagagtcc atccccacag ggtggcccag
     4501 acgatgtgag cctactcagt cctggggaag acgtgctcat cgacattgat gacaaggagc
     4561 ccctcatccc catccaggag gaagagttca tcgattggtg gagcaaattc tttgcctcca
     4621 taggggagag ggaaaagtgc ggctcctacc tggagaagga ttttgacacc ctgaaggtct
     4681 atgacacaca gctggagaat gtggaggcct ttgagggcct gtctgacttt tgtaacacct
     4741 tcaagctgta ccggggcaag acgcaggagg agacagaaga tccatctgtg attggtgaat
     4801 ttaagggcct cttcaaaatt tatcccctcc cagaagaccc agccatcccc atgcccccaa
     4861 gacagttcca ccagctggcc gcccagggac cccaggagtg cttggtccgt atctacattg
     4921 tccgagcatt tggcctgcag cccaaggacc ccaatggaaa gtgtgatcct tacatcaaga
     4981 tctccatagg gaagaaatca gtgagtgacc aggataacta catcccctgc acgctggagc
     5041 ccgtatttgg aaagatgttc gagctgacct gcactctgcc tctggagaag gacctaaaga
     5101 tcactctcta tgactatgac ctcctctcca aggacgaaaa gatcggtgag acggtcgtcg
     5161 acctggagaa caggctgctg tccaagtttg gggctcgctg tggactccca cagacctact
     5221 gtgtctctgg accgaaccag tggcgggacc agctccgccc ctcccagctc ctccacctct
     5281 tctgccagca gcatagagtc aaggcacctg tgtaccggac agaccgtgta atgtttcagg
     5341 ataaagaata ttccattgaa gagatagagg ctggcaggat cccaaaccca cacctgggcc
     5401 cagtggagga gcgtctggct ctgcatgtgc ttcagcagca gggcctggtc ccggagcacg
     5461 tggagtcacg gcccctctac agccccctgc agccagacat cgagcagggg aagctgcaga
     5521 tgtgggtcga cctatttccg aaggccctgg ggcggcctgg acctcccttc aacatcaccc
     5581 cacggagagc cagaaggttt ttcctgcgtt gtattatctg gaataccaga gatgtgatcc
     5641 tggatgacct gagcctcacg ggggagaaga tgagcgacat ttatgtgaaa ggttggatga
     5701 ttggctttga agaacacaag caaaagacag acgtgcatta tcgttccctg ggaggtgaag
     5761 gcaacttcaa ctggaggttc attttcccct tcgactacct gccagctgag caagtctgta
     5821 ccattgccaa gaaggatgcc ttctggaggc tggacaagac tgagagcaaa atcccagcac
     5881 gagtggtgtt ccagatctgg gacaatgaca agttctcctt tgatgatttt ctgggctccc
     5941 tgcagctcga tctcaaccgc atgcccaagc cagccaagac agccaagaag tgctccttgg
     6001 accagctgga tgatgctttc cacccagaat ggtttgtgtc cctttttgag cagaaaacag
     6061 tgaagggctg gtggccctgt gtagcagaag agggtgagaa gaaaatactg gcgggcaagc
     6121 tggaaatgac cttggagatt gtagcagaga gtgagcatga ggagcggcct gctggccagg
     6181 gccgggatga gcccaacatg aaccctaagc ttgaggaccc aaggcgcccc gacacctcct
     6241 tcctgtggtt tacctcccca tacaagacca tgaagttcat cctgtggcgg cgtttccggt
     6301 gggccatcat cctcttcatc atcctcttca tcctgctgct gttcctggcc atcttcatct
     6361 acgccttccc gaactatgct gccatgaagc tggtgaagcc cttcagctga tgatcaccat
     6421 ggcatatggc tagcgcggcc gccatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7261 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca
     7621 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     7681 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     7741 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     7801 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     7861 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     7921 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     7981 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     8041 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     8101 tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     8161 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     8221 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     8281 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     8341 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     8401 gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag catttatcag
     8461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8521 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8581 acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
