LOCUS       sequence-96273-e        8605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 14 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8605)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8605)
  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            "Unpublished"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-Exon14-del
FEATURES             Location/Qualifiers
     source          1..8605
                     /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      6420..6429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6429..6445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6429..6445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6442..6464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6453..6469
                     /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(6477..6483,6484..6501,6502..6507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6522..6543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6660..6677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6831..7419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6911..6930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7590..8405)
                     /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(7590..8405)
                     /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     8313..8332
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8406..8510)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8578..8596
                     /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 tgtttccctc catgtgcgaa aaaatgagga ttcgtatcat agactgggac cgcctgactc
     1561 acaatgacat cgtggctacc acctacctga gtatgtcgaa aatctctgcc cctggaggag
     1621 aaatagaaga ggagcctgca ggtgctgtca agccttcgaa agcctcagac ttggatgact
     1681 acctgggctt cctccccact tttgggccct gctacatcaa cctctatggc agtcccagag
     1741 agttcacagg cttcccagac ccctacacag agctcaacac aggcaagggg gaaggtgtgg
     1801 cttatcgtgg ccggcttctg ctctccctgg agaccaagct ggtggagcac agtgaacaga
     1861 aggtggagga ccttcctgcg gatgacatcc tccgggtgga gaagtacctt aggaggcgca
     1921 agtactccct gtttgcggcc ttctactcag ccaccatgct gcaggatgtg gatgatgcca
     1981 tccagtttga ggtcagcatc gggaactacg ggaacaagtt cgacatgacc tgcctgccgc
     2041 tggcctccac cactcagtac agccgtgcag tctttgacgg gtgccactac tactacctac
     2101 cctggggtaa cgtgaaacct gtggtggtgc tgtcatccta ctgggaggac atcagccata
     2161 gaatcgagac tcagaaccag ctgcttggga ttgctgaccg gctggaagct ggcctggagc
     2221 aggtccacct ggccctgaag gcgcagtgct ccacggagga cgtggactcg ctggtggctc
     2281 agctgacgga tgagctcatc gcaggctgca gccagcctct gggtgacatc catgagacac
     2341 cctctgccac ccacctggac cagtacctgt accagctgcg cacccatcac ctgagccaaa
     2401 tcactgaggc tgccctggcc ctgaagctcg gccacagtga gctccctgca gctctggagc
     2461 aggcggagga ctggctcctg cgtctgcgtg ccctggcaga ggagccccag aacagcctgc
     2521 cggacatcgt catctggatg ctgcagggag acaagcgtgt ggcataccag cgggtgcccg
     2581 cccaccaagt cctcttctcc cggcggggtg ccaactactg tggcaagaat tgtgggaagc
     2641 tacagacaat ctttctgaaa tatccgatgg agaaggtgcc tggcgcccgg atgccagtgc
     2701 agatacgggt caagctgtgg tttgggctct ctgtggatga gaaggagttc aaccagtttg
     2761 ctgaggggaa gctgtctgtc tttgctgaaa cctatgagaa cgagactaag ttggcccttg
     2821 ttgggaactg gggcacaacg ggcctcacct accccaagtt ttctgacgtc acgggcaaga
     2881 tcaagctacc caaggacagc ttccgcccct cggccggctg gacctgggct ggagattggt
     2941 tcgtgtgtcc ggagaagact ctgctccatg acatggacgc cggtcacctg agcttcgtgg
     3001 aagaggtgtt tgagaaccag acccggcttc ccggaggcca gtggatctac atgagtgaca
     3061 actacaccga tgtgaacggg gagaaggtgc ttcccaagga tgacattgag tgcccactgg
     3121 gctggaagtg ggaagatgag gaatggtcca cagacctcaa ccgggctgtc gatgagcaag
     3181 gctgggagta tagcatcacc atccccccgg agcggaagcc gaagcactgg gtccctgctg
     3241 agaagatgta ctacacacac cgacggcggc gctgggtgcg cctgcgcagg agggatctca
     3301 gccaaatgga agcactgaaa aggcacaggc aggcggaggc ggagggcgag ggctgggagt
     3361 acgcctctct ttttggctgg aagttccacc tcgagtaccg caagacagat gccttccgcc
     3421 gccgccgctg gcgccgtcgc atggagccac tggagaagac ggggcctgca gctgtgtttg
     3481 cccttgaggg ggccctgggc ggcgtgatgg atgacaagag tgaagattcc atgtccgtct
     3541 ccaccttgag cttcggtgtg aacagaccca cgatttcctg catattcgac tatgggaacc
     3601 gctaccatct acgctgctac atgtaccagg cccgggacct ggctgcgatg gacaaggact
     3661 ctttttctga tccctatgcc atcgtctcct tcctgcacca gagccagaag acggtggtgg
     3721 tgaagaacac ccttaacccc acctgggacc agacgctcat cttctacgag atcgagatct
     3781 ttggcgagcc ggccacagtt gctgagcaac cgcccagcat tgtggtggag ctgtacgacc
     3841 atgacactta tggtgcagac gagtttatgg gtcgctgcat ctgtcaaccg agtctggaac
     3901 ggatgccacg gctggcctgg ttcccactga cgaggggcag ccagccgtcg ggggagctgc
     3961 tggcctcttt tgagctcatc cagagagaga agccggccat ccaccatatt cctggttttg
     4021 aggtgcagga gacatcaagg atcctggatg agtctgagga cacagacctg ccctacccac
     4081 caccccagag ggaggccaac atctacatgg ttcctcagaa catcaagcca gcgctccagc
     4141 gtaccgccat cgagatcctg gcatggggcc tgcggaacat gaagagttac cagctggcca
     4201 acatctcctc ccccagcctc gtggtagagt gtgggggcca gacggtgcag tcctgtgtca
     4261 tcaggaacct ccggaagaac cccaactttg acatctgcac cctcttcatg gaagtgatgc
     4321 tgcccaggga ggagctctac tgccccccca tcaccgtcaa ggtcatcgat aaccgccagt
     4381 ttggccgccg gcctgtggtg ggccagtgta ccatccgctc cctggagagc ttcctgtgtg
     4441 acccctactc ggcggagagt ccatccccac agggtggccc agacgatgtg agcctactca
     4501 gtcctgggga agacgtgctc atcgacattg atgacaagga gcccctcatc cccatccagg
     4561 aggaagagtt catcgattgg tggagcaaat tctttgcctc cataggggag agggaaaagt
     4621 gcggctccta cctggagaag gattttgaca ccctgaaggt ctatgacaca cagctggaga
     4681 atgtggaggc ctttgagggc ctgtctgact tttgtaacac cttcaagctg taccggggca
     4741 agacgcagga ggagacagaa gatccatctg tgattggtga atttaagggc ctcttcaaaa
     4801 tttatcccct cccagaagac ccagccatcc ccatgccccc aagacagttc caccagctgg
     4861 ccgcccaggg accccaggag tgcttggtcc gtatctacat tgtccgagca tttggcctgc
     4921 agcccaagga ccccaatgga aagtgtgatc cttacatcaa gatctccata gggaagaaat
     4981 cagtgagtga ccaggataac tacatcccct gcacgctgga gcccgtattt ggaaagatgt
     5041 tcgagctgac ctgcactctg cctctggaga aggacctaaa gatcactctc tatgactatg
     5101 acctcctctc caaggacgaa aagatcggtg agacggtcgt cgacctggag aacaggctgc
     5161 tgtccaagtt tggggctcgc tgtggactcc cacagaccta ctgtgtctct ggaccgaacc
     5221 agtggcggga ccagctccgc ccctcccagc tcctccacct cttctgccag cagcatagag
     5281 tcaaggcacc tgtgtaccgg acagaccgtg taatgtttca ggataaagaa tattccattg
     5341 aagagataga ggctggcagg atcccaaacc cacacctggg cccagtggag gagcgtctgg
     5401 ctctgcatgt gcttcagcag cagggcctgg tcccggagca cgtggagtca cggcccctct
     5461 acagccccct gcagccagac atcgagcagg ggaagctgca gatgtgggtc gacctatttc
     5521 cgaaggccct ggggcggcct ggacctccct tcaacatcac cccacggaga gccagaaggt
     5581 ttttcctgcg ttgtattatc tggaatacca gagatgtgat cctggatgac ctgagcctca
     5641 cgggggagaa gatgagcgac atttatgtga aaggttggat gattggcttt gaagaacaca
     5701 agcaaaagac agacgtgcat tatcgttccc tgggaggtga aggcaacttc aactggaggt
     5761 tcattttccc cttcgactac ctgccagctg agcaagtctg taccattgcc aagaaggatg
     5821 ccttctggag gctggacaag actgagagca aaatcccagc acgagtggtg ttccagatct
     5881 gggacaatga caagttctcc tttgatgatt ttctgggctc cctgcagctc gatctcaacc
     5941 gcatgcccaa gccagccaag acagccaaga agtgctcctt ggaccagctg gatgatgctt
     6001 tccacccaga atggtttgtg tccctttttg agcagaaaac agtgaagggc tggtggccct
     6061 gtgtagcaga agagggtgag aagaaaatac tggcgggcaa gctggaaatg accttggaga
     6121 ttgtagcaga gagtgagcat gaggagcggc ctgctggcca gggccgggat gagcccaaca
     6181 tgaaccctaa gcttgaggac ccaaggcgcc ccgacacctc cttcctgtgg tttacctccc
     6241 catacaagac catgaagttc atcctgtggc ggcgtttccg gtgggccatc atcctcttca
     6301 tcatcctctt catcctgctg ctgttcctgg ccatcttcat ctacgccttc ccgaactatg
     6361 ctgccatgaa gctggtgaag cccttcagct gatgatcacc atggcatatg gctagcgcgg
     6421 ccgccatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6481 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6541 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6601 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6661 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6721 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6781 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6841 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6901 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6961 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7021 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7081 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7141 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7201 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7261 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7321 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7381 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7441 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7501 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7561 agtatatatg agtaaacttg gtctgacagt tagaaaaact catcgagcat caaatgaaac
     7621 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     7681 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     7741 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     7801 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     7861 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     7921 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     7981 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     8041 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccca
     8101 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     8161 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     8221 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     8281 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     8341 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     8401 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8461 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8521 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     8581 aggcgtatca cgaggccctt tcgtc
//
