LOCUS       sequence-96275-e        8632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 17 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8632)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8632)
  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            "Unpublished"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-Exon17-del
FEATURES             Location/Qualifiers
     source          1..8632
                     /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      6447..6456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6456..6472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6456..6472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6469..6491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6480..6496
                     /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(6504..6510,6511..6528,6529..6534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6549..6570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6687..6704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6858..7446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6938..6957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7617..8432)
                     /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(7617..8432)
                     /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     8340..8359
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8433..8537)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8605..8623
                     /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 aatagaagtg gatgactacc tgggcttcct ccccactttt gggccctgct
     1741 acatcaacct ctatggcagt cccagagagt tcacaggctt cccagacccc tacacagagc
     1801 tcaacacagg caagggggaa ggtgtggctt atcgtggccg gcttctgctc tccctggaga
     1861 ccaagctggt ggagcacagt gaacagaagg tggaggacct tcctgcggat gacatcctcc
     1921 gggtggagaa gtaccttagg aggcgcaagt actccctgtt tgcggccttc tactcagcca
     1981 ccatgctgca ggatgtggat gatgccatcc agtttgaggt cagcatcggg aactacggga
     2041 acaagttcga catgacctgc ctgccgctgg cctccaccac tcagtacagc cgtgcagtct
     2101 ttgacgggtg ccactactac tacctaccct ggggtaacgt gaaacctgtg gtggtgctgt
     2161 catcctactg ggaggacatc agccatagaa tcgagactca gaaccagctg cttgggattg
     2221 ctgaccggct ggaagctggc ctggagcagg tccacctggc cctgaaggcg cagtgctcca
     2281 cggaggacgt ggactcgctg gtggctcagc tgacggatga gctcatcgca ggctgcagcc
     2341 agcctctggg tgacatccat gagacaccct ctgccaccca cctggaccag tacctgtacc
     2401 agctgcgcac ccatcacctg agccaaatca ctgaggctgc cctggccctg aagctcggcc
     2461 acagtgagct ccctgcagct ctggagcagg cggaggactg gctcctgcgt ctgcgtgccc
     2521 tggcagagga gccccagaac agcctgccgg acatcgtcat ctggatgctg cagggagaca
     2581 agcgtgtggc ataccagcgg gtgcccgccc accaagtcct cttctcccgg cggggtgcca
     2641 actactgtgg caagaattgt gggaagctac agacaatctt tctgaaatat ccgatggaga
     2701 aggtgcctgg cgcccggatg ccagtgcaga tacgggtcaa gctgtggttt gggctctctg
     2761 tggatgagaa ggagttcaac cagtttgctg aggggaagct gtctgtcttt gctgaaacct
     2821 atgagaacga gactaagttg gcccttgttg ggaactgggg cacaacgggc ctcacctacc
     2881 ccaagttttc tgacgtcacg ggcaagatca agctacccaa ggacagcttc cgcccctcgg
     2941 ccggctggac ctgggctgga gattggttcg tgtgtccgga gaagactctg ctccatgaca
     3001 tggacgccgg tcacctgagc ttcgtggaag aggtgtttga gaaccagacc cggcttcccg
     3061 gaggccagtg gatctacatg agtgacaact acaccgatgt gaacggggag aaggtgcttc
     3121 ccaaggatga cattgagtgc ccactgggct ggaagtggga agatgaggaa tggtccacag
     3181 acctcaaccg ggctgtcgat gagcaaggct gggagtatag catcaccatc cccccggagc
     3241 ggaagccgaa gcactgggtc cctgctgaga agatgtacta cacacaccga cggcggcgct
     3301 gggtgcgcct gcgcaggagg gatctcagcc aaatggaagc actgaaaagg cacaggcagg
     3361 cggaggcgga gggcgagggc tgggagtacg cctctctttt tggctggaag ttccacctcg
     3421 agtaccgcaa gacagatgcc ttccgccgcc gccgctggcg ccgtcgcatg gagccactgg
     3481 agaagacggg gcctgcagct gtgtttgccc ttgagggggc cctgggcggc gtgatggatg
     3541 acaagagtga agattccatg tccgtctcca ccttgagctt cggtgtgaac agacccacga
     3601 tttcctgcat attcgactat gggaaccgct accatctacg ctgctacatg taccaggccc
     3661 gggacctggc tgcgatggac aaggactctt tttctgatcc ctatgccatc gtctccttcc
     3721 tgcaccagag ccagaagacg gtggtggtga agaacaccct taaccccacc tgggaccaga
     3781 cgctcatctt ctacgagatc gagatctttg gcgagccggc cacagttgct gagcaaccgc
     3841 ccagcattgt ggtggagctg tacgaccatg acacttatgg tgcagacgag tttatgggtc
     3901 gctgcatctg tcaaccgagt ctggaacgga tgccacggct ggcctggttc ccactgacga
     3961 ggggcagcca gccgtcgggg gagctgctgg cctcttttga gctcatccag agagagaagc
     4021 cggccatcca ccatattcct ggttttgagg tgcaggagac atcaaggatc ctggatgagt
     4081 ctgaggacac agacctgccc tacccaccac cccagaggga ggccaacatc tacatggttc
     4141 ctcagaacat caagccagcg ctccagcgta ccgccatcga gatcctggca tggggcctgc
     4201 ggaacatgaa gagttaccag ctggccaaca tctcctcccc cagcctcgtg gtagagtgtg
     4261 ggggccagac ggtgcagtcc tgtgtcatca ggaacctccg gaagaacccc aactttgaca
     4321 tctgcaccct cttcatggaa gtgatgctgc ccagggagga gctctactgc ccccccatca
     4381 ccgtcaaggt catcgataac cgccagtttg gccgccggcc tgtggtgggc cagtgtacca
     4441 tccgctccct ggagagcttc ctgtgtgacc cctactcggc ggagagtcca tccccacagg
     4501 gtggcccaga cgatgtgagc ctactcagtc ctggggaaga cgtgctcatc gacattgatg
     4561 acaaggagcc cctcatcccc atccaggagg aagagttcat cgattggtgg agcaaattct
     4621 ttgcctccat aggggagagg gaaaagtgcg gctcctacct ggagaaggat tttgacaccc
     4681 tgaaggtcta tgacacacag ctggagaatg tggaggcctt tgagggcctg tctgactttt
     4741 gtaacacctt caagctgtac cggggcaaga cgcaggagga gacagaagat ccatctgtga
     4801 ttggtgaatt taagggcctc ttcaaaattt atcccctccc agaagaccca gccatcccca
     4861 tgcccccaag acagttccac cagctggccg cccagggacc ccaggagtgc ttggtccgta
     4921 tctacattgt ccgagcattt ggcctgcagc ccaaggaccc caatggaaag tgtgatcctt
     4981 acatcaagat ctccataggg aagaaatcag tgagtgacca ggataactac atcccctgca
     5041 cgctggagcc cgtatttgga aagatgttcg agctgacctg cactctgcct ctggagaagg
     5101 acctaaagat cactctctat gactatgacc tcctctccaa ggacgaaaag atcggtgaga
     5161 cggtcgtcga cctggagaac aggctgctgt ccaagtttgg ggctcgctgt ggactcccac
     5221 agacctactg tgtctctgga ccgaaccagt ggcgggacca gctccgcccc tcccagctcc
     5281 tccacctctt ctgccagcag catagagtca aggcacctgt gtaccggaca gaccgtgtaa
     5341 tgtttcagga taaagaatat tccattgaag agatagaggc tggcaggatc ccaaacccac
     5401 acctgggccc agtggaggag cgtctggctc tgcatgtgct tcagcagcag ggcctggtcc
     5461 cggagcacgt ggagtcacgg cccctctaca gccccctgca gccagacatc gagcagggga
     5521 agctgcagat gtgggtcgac ctatttccga aggccctggg gcggcctgga cctcccttca
     5581 acatcacccc acggagagcc agaaggtttt tcctgcgttg tattatctgg aataccagag
     5641 atgtgatcct ggatgacctg agcctcacgg gggagaagat gagcgacatt tatgtgaaag
     5701 gttggatgat tggctttgaa gaacacaagc aaaagacaga cgtgcattat cgttccctgg
     5761 gaggtgaagg caacttcaac tggaggttca ttttcccctt cgactacctg ccagctgagc
     5821 aagtctgtac cattgccaag aaggatgcct tctggaggct ggacaagact gagagcaaaa
     5881 tcccagcacg agtggtgttc cagatctggg acaatgacaa gttctccttt gatgattttc
     5941 tgggctccct gcagctcgat ctcaaccgca tgcccaagcc agccaagaca gccaagaagt
     6001 gctccttgga ccagctggat gatgctttcc acccagaatg gtttgtgtcc ctttttgagc
     6061 agaaaacagt gaagggctgg tggccctgtg tagcagaaga gggtgagaag aaaatactgg
     6121 cgggcaagct ggaaatgacc ttggagattg tagcagagag tgagcatgag gagcggcctg
     6181 ctggccaggg ccgggatgag cccaacatga accctaagct tgaggaccca aggcgccccg
     6241 acacctcctt cctgtggttt acctccccat acaagaccat gaagttcatc ctgtggcggc
     6301 gtttccggtg ggccatcatc ctcttcatca tcctcttcat cctgctgctg ttcctggcca
     6361 tcttcatcta cgccttcccg aactatgctg ccatgaagct ggtgaagccc ttcagctgat
     6421 gatcaccatg gcatatggct agcgcggccg ccatggtcat agctgtttcc tgtgtgaaat
     6481 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6541 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6601 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6661 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6721 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6781 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6901 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7021 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7261 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7321 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7381 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7441 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7501 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7561 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttag
     7621 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     7681 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     7741 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     7801 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     7861 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     7921 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     7981 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     8041 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     8101 tctaatacct ggaatgctgt tttcccaggg atcgcagtgg tgagtaacca tgcatcatca
     8161 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     8221 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     8281 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     8341 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     8401 gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata ttattgaagc
     8461 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8521 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8581 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
