LOCUS       sequence-96280-e        8542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 25 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8542)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8542)
  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            "Unpublished"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-Exon25-del
FEATURES             Location/Qualifiers
     source          1..8542
                     /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      6357..6366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6366..6382)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6366..6382)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6379..6401)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6390..6406
                     /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(6414..6420,6421..6438,6439..6444))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6459..6480
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6597..6614)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6768..7356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6848..6867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7527..8342)
                     /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(7527..8342)
                     /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     8250..8269
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8343..8447)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8515..8533
                     /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 atgagaacga gactaagttg gcccttgttg
     2761 ggaactgggg cacaacgggc ctcacctacc ccaagttttc tgacgtcacg ggcaagatca
     2821 agctacccaa ggacagcttc cgcccctcgg ccggctggac ctgggctgga gattggttcg
     2881 tgtgtccgga gaagactctg ctccatgaca tggacgccgg tcacctgagc ttcgtggaag
     2941 aggtgtttga gaaccagacc cggcttcccg gaggccagtg gatctacatg agtgacaact
     3001 acaccgatgt gaacggggag aaggtgcttc ccaaggatga cattgagtgc ccactgggct
     3061 ggaagtggga agatgaggaa tggtccacag acctcaaccg ggctgtcgat gagcaaggct
     3121 gggagtatag catcaccatc cccccggagc ggaagccgaa gcactgggtc cctgctgaga
     3181 agatgtacta cacacaccga cggcggcgct gggtgcgcct gcgcaggagg gatctcagcc
     3241 aaatggaagc actgaaaagg cacaggcagg cggaggcgga gggcgagggc tgggagtacg
     3301 cctctctttt tggctggaag ttccacctcg agtaccgcaa gacagatgcc ttccgccgcc
     3361 gccgctggcg ccgtcgcatg gagccactgg agaagacggg gcctgcagct gtgtttgccc
     3421 ttgagggggc cctgggcggc gtgatggatg acaagagtga agattccatg tccgtctcca
     3481 ccttgagctt cggtgtgaac agacccacga tttcctgcat attcgactat gggaaccgct
     3541 accatctacg ctgctacatg taccaggccc gggacctggc tgcgatggac aaggactctt
     3601 tttctgatcc ctatgccatc gtctccttcc tgcaccagag ccagaagacg gtggtggtga
     3661 agaacaccct taaccccacc tgggaccaga cgctcatctt ctacgagatc gagatctttg
     3721 gcgagccggc cacagttgct gagcaaccgc ccagcattgt ggtggagctg tacgaccatg
     3781 acacttatgg tgcagacgag tttatgggtc gctgcatctg tcaaccgagt ctggaacgga
     3841 tgccacggct ggcctggttc ccactgacga ggggcagcca gccgtcgggg gagctgctgg
     3901 cctcttttga gctcatccag agagagaagc cggccatcca ccatattcct ggttttgagg
     3961 tgcaggagac atcaaggatc ctggatgagt ctgaggacac agacctgccc tacccaccac
     4021 cccagaggga ggccaacatc tacatggttc ctcagaacat caagccagcg ctccagcgta
     4081 ccgccatcga gatcctggca tggggcctgc ggaacatgaa gagttaccag ctggccaaca
     4141 tctcctcccc cagcctcgtg gtagagtgtg ggggccagac ggtgcagtcc tgtgtcatca
     4201 ggaacctccg gaagaacccc aactttgaca tctgcaccct cttcatggaa gtgatgctgc
     4261 ccagggagga gctctactgc ccccccatca ccgtcaaggt catcgataac cgccagtttg
     4321 gccgccggcc tgtggtgggc cagtgtacca tccgctccct ggagagcttc ctgtgtgacc
     4381 cctactcggc ggagagtcca tccccacagg gtggcccaga cgatgtgagc ctactcagtc
     4441 ctggggaaga cgtgctcatc gacattgatg acaaggagcc cctcatcccc atccaggagg
     4501 aagagttcat cgattggtgg agcaaattct ttgcctccat aggggagagg gaaaagtgcg
     4561 gctcctacct ggagaaggat tttgacaccc tgaaggtcta tgacacacag ctggagaatg
     4621 tggaggcctt tgagggcctg tctgactttt gtaacacctt caagctgtac cggggcaaga
     4681 cgcaggagga gacagaagat ccatctgtga ttggtgaatt taagggcctc ttcaaaattt
     4741 atcccctccc agaagaccca gccatcccca tgcccccaag acagttccac cagctggccg
     4801 cccagggacc ccaggagtgc ttggtccgta tctacattgt ccgagcattt ggcctgcagc
     4861 ccaaggaccc caatggaaag tgtgatcctt acatcaagat ctccataggg aagaaatcag
     4921 tgagtgacca ggataactac atcccctgca cgctggagcc cgtatttgga aagatgttcg
     4981 agctgacctg cactctgcct ctggagaagg acctaaagat cactctctat gactatgacc
     5041 tcctctccaa ggacgaaaag atcggtgaga cggtcgtcga cctggagaac aggctgctgt
     5101 ccaagtttgg ggctcgctgt ggactcccac agacctactg tgtctctgga ccgaaccagt
     5161 ggcgggacca gctccgcccc tcccagctcc tccacctctt ctgccagcag catagagtca
     5221 aggcacctgt gtaccggaca gaccgtgtaa tgtttcagga taaagaatat tccattgaag
     5281 agatagaggc tggcaggatc ccaaacccac acctgggccc agtggaggag cgtctggctc
     5341 tgcatgtgct tcagcagcag ggcctggtcc cggagcacgt ggagtcacgg cccctctaca
     5401 gccccctgca gccagacatc gagcagggga agctgcagat gtgggtcgac ctatttccga
     5461 aggccctggg gcggcctgga cctcccttca acatcacccc acggagagcc agaaggtttt
     5521 tcctgcgttg tattatctgg aataccagag atgtgatcct ggatgacctg agcctcacgg
     5581 gggagaagat gagcgacatt tatgtgaaag gttggatgat tggctttgaa gaacacaagc
     5641 aaaagacaga cgtgcattat cgttccctgg gaggtgaagg caacttcaac tggaggttca
     5701 ttttcccctt cgactacctg ccagctgagc aagtctgtac cattgccaag aaggatgcct
     5761 tctggaggct ggacaagact gagagcaaaa tcccagcacg agtggtgttc cagatctggg
     5821 acaatgacaa gttctccttt gatgattttc tgggctccct gcagctcgat ctcaaccgca
     5881 tgcccaagcc agccaagaca gccaagaagt gctccttgga ccagctggat gatgctttcc
     5941 acccagaatg gtttgtgtcc ctttttgagc agaaaacagt gaagggctgg tggccctgtg
     6001 tagcagaaga gggtgagaag aaaatactgg cgggcaagct ggaaatgacc ttggagattg
     6061 tagcagagag tgagcatgag gagcggcctg ctggccaggg ccgggatgag cccaacatga
     6121 accctaagct tgaggaccca aggcgccccg acacctcctt cctgtggttt acctccccat
     6181 acaagaccat gaagttcatc ctgtggcggc gtttccggtg ggccatcatc ctcttcatca
     6241 tcctcttcat cctgctgctg ttcctggcca tcttcatcta cgccttcccg aactatgctg
     6301 ccatgaagct ggtgaagccc ttcagctgat gatcaccatg gcatatggct agcgcggccg
     6361 ccatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6421 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6481 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6541 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6601 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6661 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6721 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6781 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6841 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6901 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6961 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7021 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7081 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7141 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7201 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7261 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7321 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7381 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7441 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7501 atatatgagt aaacttggtc tgacagttag aaaaactcat cgagcatcaa atgaaactgc
     7561 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     7621 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     7681 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     7741 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     7801 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     7861 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     7921 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     7981 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccaggg
     8041 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     8101 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     8161 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     8221 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     8281 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     8341 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8401 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8461 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     8521 cgtatcacga ggccctttcg tc
//
