LOCUS       sequence-96281-e        8500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 30 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8500)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8500)
  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            "Unpublished"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-Exon30-del
FEATURES             Location/Qualifiers
     source          1..8500
                     /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      6315..6324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6324..6340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6324..6340)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6337..6359)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6348..6364
                     /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(6372..6378,6379..6396,6397..6402))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6417..6438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6555..6572)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6726..7314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6806..6825)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7485..8300)
                     /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(7485..8300)
                     /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     8208..8227
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8301..8405)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8473..8491
                     /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 ggggcggcgt gatggatgac aagagtgaag
     3421 attccatgtc cgtctccacc ttgagcttcg gtgtgaacag acccacgatt tcctgcatat
     3481 tcgactatgg gaaccgctac catctacgct gctacatgta ccaggcccgg gacctggctg
     3541 cgatggacaa ggactctttt tctgatccct atgccatcgt ctccttcctg caccagagcc
     3601 agaagacggt ggtggtgaag aacaccctta accccacctg ggaccagacg ctcatcttct
     3661 acgagatcga gatctttggc gagccggcca cagttgctga gcaaccgccc agcattgtgg
     3721 tggagctgta cgaccatgac acttatggtg cagacgagtt tatgggtcgc tgcatctgtc
     3781 aaccgagtct ggaacggatg ccacggctgg cctggttccc actgacgagg ggcagccagc
     3841 cgtcggggga gctgctggcc tcttttgagc tcatccagag agagaagccg gccatccacc
     3901 atattcctgg ttttgaggtg caggagacat caaggatcct ggatgagtct gaggacacag
     3961 acctgcccta cccaccaccc cagagggagg ccaacatcta catggttcct cagaacatca
     4021 agccagcgct ccagcgtacc gccatcgaga tcctggcatg gggcctgcgg aacatgaaga
     4081 gttaccagct ggccaacatc tcctccccca gcctcgtggt agagtgtggg ggccagacgg
     4141 tgcagtcctg tgtcatcagg aacctccgga agaaccccaa ctttgacatc tgcaccctct
     4201 tcatggaagt gatgctgccc agggaggagc tctactgccc ccccatcacc gtcaaggtca
     4261 tcgataaccg ccagtttggc cgccggcctg tggtgggcca gtgtaccatc cgctccctgg
     4321 agagcttcct gtgtgacccc tactcggcgg agagtccatc cccacagggt ggcccagacg
     4381 atgtgagcct actcagtcct ggggaagacg tgctcatcga cattgatgac aaggagcccc
     4441 tcatccccat ccaggaggaa gagttcatcg attggtggag caaattcttt gcctccatag
     4501 gggagaggga aaagtgcggc tcctacctgg agaaggattt tgacaccctg aaggtctatg
     4561 acacacagct ggagaatgtg gaggcctttg agggcctgtc tgacttttgt aacaccttca
     4621 agctgtaccg gggcaagacg caggaggaga cagaagatcc atctgtgatt ggtgaattta
     4681 agggcctctt caaaatttat cccctcccag aagacccagc catccccatg cccccaagac
     4741 agttccacca gctggccgcc cagggacccc aggagtgctt ggtccgtatc tacattgtcc
     4801 gagcatttgg cctgcagccc aaggacccca atggaaagtg tgatccttac atcaagatct
     4861 ccatagggaa gaaatcagtg agtgaccagg ataactacat cccctgcacg ctggagcccg
     4921 tatttggaaa gatgttcgag ctgacctgca ctctgcctct ggagaaggac ctaaagatca
     4981 ctctctatga ctatgacctc ctctccaagg acgaaaagat cggtgagacg gtcgtcgacc
     5041 tggagaacag gctgctgtcc aagtttgggg ctcgctgtgg actcccacag acctactgtg
     5101 tctctggacc gaaccagtgg cgggaccagc tccgcccctc ccagctcctc cacctcttct
     5161 gccagcagca tagagtcaag gcacctgtgt accggacaga ccgtgtaatg tttcaggata
     5221 aagaatattc cattgaagag atagaggctg gcaggatccc aaacccacac ctgggcccag
     5281 tggaggagcg tctggctctg catgtgcttc agcagcaggg cctggtcccg gagcacgtgg
     5341 agtcacggcc cctctacagc cccctgcagc cagacatcga gcaggggaag ctgcagatgt
     5401 gggtcgacct atttccgaag gccctggggc ggcctggacc tcccttcaac atcaccccac
     5461 ggagagccag aaggtttttc ctgcgttgta ttatctggaa taccagagat gtgatcctgg
     5521 atgacctgag cctcacgggg gagaagatga gcgacattta tgtgaaaggt tggatgattg
     5581 gctttgaaga acacaagcaa aagacagacg tgcattatcg ttccctggga ggtgaaggca
     5641 acttcaactg gaggttcatt ttccccttcg actacctgcc agctgagcaa gtctgtacca
     5701 ttgccaagaa ggatgccttc tggaggctgg acaagactga gagcaaaatc ccagcacgag
     5761 tggtgttcca gatctgggac aatgacaagt tctcctttga tgattttctg ggctccctgc
     5821 agctcgatct caaccgcatg cccaagccag ccaagacagc caagaagtgc tccttggacc
     5881 agctggatga tgctttccac ccagaatggt ttgtgtccct ttttgagcag aaaacagtga
     5941 agggctggtg gccctgtgta gcagaagagg gtgagaagaa aatactggcg ggcaagctgg
     6001 aaatgacctt ggagattgta gcagagagtg agcatgagga gcggcctgct ggccagggcc
     6061 gggatgagcc caacatgaac cctaagcttg aggacccaag gcgccccgac acctccttcc
     6121 tgtggtttac ctccccatac aagaccatga agttcatcct gtggcggcgt ttccggtggg
     6181 ccatcatcct cttcatcatc ctcttcatcc tgctgctgtt cctggccatc ttcatctacg
     6241 ccttcccgaa ctatgctgcc atgaagctgg tgaagccctt cagctgatga tcaccatggc
     6301 atatggctag cgcggccgcc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6361 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6421 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6481 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6541 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6601 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6661 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6721 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6781 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6841 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6901 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6961 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7021 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7081 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7141 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7201 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7261 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7321 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7381 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7441 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttagaa aaactcatcg
     7501 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     7561 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     7621 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     7681 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     7741 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     7801 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     7861 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     7921 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     7981 aatgctgttt tcccagggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     8041 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     8101 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     8161 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     8221 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     8281 tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8341 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8401 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8461 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
