LOCUS       sequence-96278-e        8560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 19 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8560)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8560)
  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            "Unpublished"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-Exon19-del
FEATURES             Location/Qualifiers
     source          1..8560
                     /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      6375..6384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6384..6400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6384..6400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6397..6419)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6408..6424
                     /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(6432..6438,6439..6456,6457..6462))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6477..6498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6615..6632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6786..7374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6866..6885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7545..8360)
                     /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(7545..8360)
                     /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     8268..8287
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8361..8465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8533..8551
                     /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 ggcaagaagt
     1861 accttaggag gcgcaagtac tccctgtttg cggccttcta ctcagccacc atgctgcagg
     1921 atgtggatga tgccatccag tttgaggtca gcatcgggaa ctacgggaac aagttcgaca
     1981 tgacctgcct gccgctggcc tccaccactc agtacagccg tgcagtcttt gacgggtgcc
     2041 actactacta cctaccctgg ggtaacgtga aacctgtggt ggtgctgtca tcctactggg
     2101 aggacatcag ccatagaatc gagactcaga accagctgct tgggattgct gaccggctgg
     2161 aagctggcct ggagcaggtc cacctggccc tgaaggcgca gtgctccacg gaggacgtgg
     2221 actcgctggt ggctcagctg acggatgagc tcatcgcagg ctgcagccag cctctgggtg
     2281 acatccatga gacaccctct gccacccacc tggaccagta cctgtaccag ctgcgcaccc
     2341 atcacctgag ccaaatcact gaggctgccc tggccctgaa gctcggccac agtgagctcc
     2401 ctgcagctct ggagcaggcg gaggactggc tcctgcgtct gcgtgccctg gcagaggagc
     2461 cccagaacag cctgccggac atcgtcatct ggatgctgca gggagacaag cgtgtggcat
     2521 accagcgggt gcccgcccac caagtcctct tctcccggcg gggtgccaac tactgtggca
     2581 agaattgtgg gaagctacag acaatctttc tgaaatatcc gatggagaag gtgcctggcg
     2641 cccggatgcc agtgcagata cgggtcaagc tgtggtttgg gctctctgtg gatgagaagg
     2701 agttcaacca gtttgctgag gggaagctgt ctgtctttgc tgaaacctat gagaacgaga
     2761 ctaagttggc ccttgttggg aactggggca caacgggcct cacctacccc aagttttctg
     2821 acgtcacggg caagatcaag ctacccaagg acagcttccg cccctcggcc ggctggacct
     2881 gggctggaga ttggttcgtg tgtccggaga agactctgct ccatgacatg gacgccggtc
     2941 acctgagctt cgtggaagag gtgtttgaga accagacccg gcttcccgga ggccagtgga
     3001 tctacatgag tgacaactac accgatgtga acggggagaa ggtgcttccc aaggatgaca
     3061 ttgagtgccc actgggctgg aagtgggaag atgaggaatg gtccacagac ctcaaccggg
     3121 ctgtcgatga gcaaggctgg gagtatagca tcaccatccc cccggagcgg aagccgaagc
     3181 actgggtccc tgctgagaag atgtactaca cacaccgacg gcggcgctgg gtgcgcctgc
     3241 gcaggaggga tctcagccaa atggaagcac tgaaaaggca caggcaggcg gaggcggagg
     3301 gcgagggctg ggagtacgcc tctctttttg gctggaagtt ccacctcgag taccgcaaga
     3361 cagatgcctt ccgccgccgc cgctggcgcc gtcgcatgga gccactggag aagacggggc
     3421 ctgcagctgt gtttgccctt gagggggccc tgggcggcgt gatggatgac aagagtgaag
     3481 attccatgtc cgtctccacc ttgagcttcg gtgtgaacag acccacgatt tcctgcatat
     3541 tcgactatgg gaaccgctac catctacgct gctacatgta ccaggcccgg gacctggctg
     3601 cgatggacaa ggactctttt tctgatccct atgccatcgt ctccttcctg caccagagcc
     3661 agaagacggt ggtggtgaag aacaccctta accccacctg ggaccagacg ctcatcttct
     3721 acgagatcga gatctttggc gagccggcca cagttgctga gcaaccgccc agcattgtgg
     3781 tggagctgta cgaccatgac acttatggtg cagacgagtt tatgggtcgc tgcatctgtc
     3841 aaccgagtct ggaacggatg ccacggctgg cctggttccc actgacgagg ggcagccagc
     3901 cgtcggggga gctgctggcc tcttttgagc tcatccagag agagaagccg gccatccacc
     3961 atattcctgg ttttgaggtg caggagacat caaggatcct ggatgagtct gaggacacag
     4021 acctgcccta cccaccaccc cagagggagg ccaacatcta catggttcct cagaacatca
     4081 agccagcgct ccagcgtacc gccatcgaga tcctggcatg gggcctgcgg aacatgaaga
     4141 gttaccagct ggccaacatc tcctccccca gcctcgtggt agagtgtggg ggccagacgg
     4201 tgcagtcctg tgtcatcagg aacctccgga agaaccccaa ctttgacatc tgcaccctct
     4261 tcatggaagt gatgctgccc agggaggagc tctactgccc ccccatcacc gtcaaggtca
     4321 tcgataaccg ccagtttggc cgccggcctg tggtgggcca gtgtaccatc cgctccctgg
     4381 agagcttcct gtgtgacccc tactcggcgg agagtccatc cccacagggt ggcccagacg
     4441 atgtgagcct actcagtcct ggggaagacg tgctcatcga cattgatgac aaggagcccc
     4501 tcatccccat ccaggaggaa gagttcatcg attggtggag caaattcttt gcctccatag
     4561 gggagaggga aaagtgcggc tcctacctgg agaaggattt tgacaccctg aaggtctatg
     4621 acacacagct ggagaatgtg gaggcctttg agggcctgtc tgacttttgt aacaccttca
     4681 agctgtaccg gggcaagacg caggaggaga cagaagatcc atctgtgatt ggtgaattta
     4741 agggcctctt caaaatttat cccctcccag aagacccagc catccccatg cccccaagac
     4801 agttccacca gctggccgcc cagggacccc aggagtgctt ggtccgtatc tacattgtcc
     4861 gagcatttgg cctgcagccc aaggacccca atggaaagtg tgatccttac atcaagatct
     4921 ccatagggaa gaaatcagtg agtgaccagg ataactacat cccctgcacg ctggagcccg
     4981 tatttggaaa gatgttcgag ctgacctgca ctctgcctct ggagaaggac ctaaagatca
     5041 ctctctatga ctatgacctc ctctccaagg acgaaaagat cggtgagacg gtcgtcgacc
     5101 tggagaacag gctgctgtcc aagtttgggg ctcgctgtgg actcccacag acctactgtg
     5161 tctctggacc gaaccagtgg cgggaccagc tccgcccctc ccagctcctc cacctcttct
     5221 gccagcagca tagagtcaag gcacctgtgt accggacaga ccgtgtaatg tttcaggata
     5281 aagaatattc cattgaagag atagaggctg gcaggatccc aaacccacac ctgggcccag
     5341 tggaggagcg tctggctctg catgtgcttc agcagcaggg cctggtcccg gagcacgtgg
     5401 agtcacggcc cctctacagc cccctgcagc cagacatcga gcaggggaag ctgcagatgt
     5461 gggtcgacct atttccgaag gccctggggc ggcctggacc tcccttcaac atcaccccac
     5521 ggagagccag aaggtttttc ctgcgttgta ttatctggaa taccagagat gtgatcctgg
     5581 atgacctgag cctcacgggg gagaagatga gcgacattta tgtgaaaggt tggatgattg
     5641 gctttgaaga acacaagcaa aagacagacg tgcattatcg ttccctggga ggtgaaggca
     5701 acttcaactg gaggttcatt ttccccttcg actacctgcc agctgagcaa gtctgtacca
     5761 ttgccaagaa ggatgccttc tggaggctgg acaagactga gagcaaaatc ccagcacgag
     5821 tggtgttcca gatctgggac aatgacaagt tctcctttga tgattttctg ggctccctgc
     5881 agctcgatct caaccgcatg cccaagccag ccaagacagc caagaagtgc tccttggacc
     5941 agctggatga tgctttccac ccagaatggt ttgtgtccct ttttgagcag aaaacagtga
     6001 agggctggtg gccctgtgta gcagaagagg gtgagaagaa aatactggcg ggcaagctgg
     6061 aaatgacctt ggagattgta gcagagagtg agcatgagga gcggcctgct ggccagggcc
     6121 gggatgagcc caacatgaac cctaagcttg aggacccaag gcgccccgac acctccttcc
     6181 tgtggtttac ctccccatac aagaccatga agttcatcct gtggcggcgt ttccggtggg
     6241 ccatcatcct cttcatcatc ctcttcatcc tgctgctgtt cctggccatc ttcatctacg
     6301 ccttcccgaa ctatgctgcc atgaagctgg tgaagccctt cagctgatga tcaccatggc
     6361 atatggctag cgcggccgcc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6421 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6481 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6601 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6661 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6721 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6781 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6841 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6901 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6961 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7021 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7081 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7141 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7201 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7261 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7321 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7381 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7441 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7501 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttagaa aaactcatcg
     7561 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     7621 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     7681 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     7741 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     7801 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     7861 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     7921 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     7981 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     8041 aatgctgttt tcccagggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     8101 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     8161 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     8221 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     8281 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     8341 tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8401 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8461 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8521 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
