LOCUS       sequence-96257-e        8674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains full-length human dysferlin in a multiple cloning 
            site of pUC57-Kan and can be easily cloned into other vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8674)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8674)
  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            "Unpublished"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: FL-DYSF-pUC57
FEATURES             Location/Qualifiers
     source          1..8674
                     /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      6489..6498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6498..6514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6498..6514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6511..6533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6522..6538
                     /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(6546..6552,6553..6570,6571..6576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6591..6612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6729..6746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6900..7488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6980..6999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7659..8474)
                     /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(7659..8474)
                     /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     8382..8401
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8475..8579)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8647..8665
                     /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 ggcacaggca ggcggaggcg gagggcgagg
     3421 gctgggagta cgcctctctt tttggctgga agttccacct cgagtaccgc aagacagatg
     3481 ccttccgccg ccgccgctgg cgccgtcgca tggagccact ggagaagacg gggcctgcag
     3541 ctgtgtttgc ccttgagggg gccctgggcg gcgtgatgga tgacaagagt gaagattcca
     3601 tgtccgtctc caccttgagc ttcggtgtga acagacccac gatttcctgc atattcgact
     3661 atgggaaccg ctaccatcta cgctgctaca tgtaccaggc ccgggacctg gctgcgatgg
     3721 acaaggactc tttttctgat ccctatgcca tcgtctcctt cctgcaccag agccagaaga
     3781 cggtggtggt gaagaacacc cttaacccca cctgggacca gacgctcatc ttctacgaga
     3841 tcgagatctt tggcgagccg gccacagttg ctgagcaacc gcccagcatt gtggtggagc
     3901 tgtacgacca tgacacttat ggtgcagacg agtttatggg tcgctgcatc tgtcaaccga
     3961 gtctggaacg gatgccacgg ctggcctggt tcccactgac gaggggcagc cagccgtcgg
     4021 gggagctgct ggcctctttt gagctcatcc agagagagaa gccggccatc caccatattc
     4081 ctggttttga ggtgcaggag acatcaagga tcctggatga gtctgaggac acagacctgc
     4141 cctacccacc accccagagg gaggccaaca tctacatggt tcctcagaac atcaagccag
     4201 cgctccagcg taccgccatc gagatcctgg catggggcct gcggaacatg aagagttacc
     4261 agctggccaa catctcctcc cccagcctcg tggtagagtg tgggggccag acggtgcagt
     4321 cctgtgtcat caggaacctc cggaagaacc ccaactttga catctgcacc ctcttcatgg
     4381 aagtgatgct gcccagggag gagctctact gcccccccat caccgtcaag gtcatcgata
     4441 accgccagtt tggccgccgg cctgtggtgg gccagtgtac catccgctcc ctggagagct
     4501 tcctgtgtga cccctactcg gcggagagtc catccccaca gggtggccca gacgatgtga
     4561 gcctactcag tcctggggaa gacgtgctca tcgacattga tgacaaggag cccctcatcc
     4621 ccatccagga ggaagagttc atcgattggt ggagcaaatt ctttgcctcc ataggggaga
     4681 gggaaaagtg cggctcctac ctggagaagg attttgacac cctgaaggtc tatgacacac
     4741 agctggagaa tgtggaggcc tttgagggcc tgtctgactt ttgtaacacc ttcaagctgt
     4801 accggggcaa gacgcaggag gagacagaag atccatctgt gattggtgaa tttaagggcc
     4861 tcttcaaaat ttatcccctc ccagaagacc cagccatccc catgccccca agacagttcc
     4921 accagctggc cgcccaggga ccccaggagt gcttggtccg tatctacatt gtccgagcat
     4981 ttggcctgca gcccaaggac cccaatggaa agtgtgatcc ttacatcaag atctccatag
     5041 ggaagaaatc agtgagtgac caggataact acatcccctg cacgctggag cccgtatttg
     5101 gaaagatgtt cgagctgacc tgcactctgc ctctggagaa ggacctaaag atcactctct
     5161 atgactatga cctcctctcc aaggacgaaa agatcggtga gacggtcgtc gacctggaga
     5221 acaggctgct gtccaagttt ggggctcgct gtggactccc acagacctac tgtgtctctg
     5281 gaccgaacca gtggcgggac cagctccgcc cctcccagct cctccacctc ttctgccagc
     5341 agcatagagt caaggcacct gtgtaccgga cagaccgtgt aatgtttcag gataaagaat
     5401 attccattga agagatagag gctggcagga tcccaaaccc acacctgggc ccagtggagg
     5461 agcgtctggc tctgcatgtg cttcagcagc agggcctggt cccggagcac gtggagtcac
     5521 ggcccctcta cagccccctg cagccagaca tcgagcaggg gaagctgcag atgtgggtcg
     5581 acctatttcc gaaggccctg gggcggcctg gacctccctt caacatcacc ccacggagag
     5641 ccagaaggtt tttcctgcgt tgtattatct ggaataccag agatgtgatc ctggatgacc
     5701 tgagcctcac gggggagaag atgagcgaca tttatgtgaa aggttggatg attggctttg
     5761 aagaacacaa gcaaaagaca gacgtgcatt atcgttccct gggaggtgaa ggcaacttca
     5821 actggaggtt cattttcccc ttcgactacc tgccagctga gcaagtctgt accattgcca
     5881 agaaggatgc cttctggagg ctggacaaga ctgagagcaa aatcccagca cgagtggtgt
     5941 tccagatctg ggacaatgac aagttctcct ttgatgattt tctgggctcc ctgcagctcg
     6001 atctcaaccg catgcccaag ccagccaaga cagccaagaa gtgctccttg gaccagctgg
     6061 atgatgcttt ccacccagaa tggtttgtgt ccctttttga gcagaaaaca gtgaagggct
     6121 ggtggccctg tgtagcagaa gagggtgaga agaaaatact ggcgggcaag ctggaaatga
     6181 ccttggagat tgtagcagag agtgagcatg aggagcggcc tgctggccag ggccgggatg
     6241 agcccaacat gaaccctaag cttgaggacc caaggcgccc cgacacctcc ttcctgtggt
     6301 ttacctcccc atacaagacc atgaagttca tcctgtggcg gcgtttccgg tgggccatca
     6361 tcctcttcat catcctcttc atcctgctgc tgttcctggc catcttcatc tacgccttcc
     6421 cgaactatgc tgccatgaag ctggtgaagc ccttcagctg atgatcacca tggcatatgg
     6481 ctagcgcggc cgccatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6541 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6661 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6721 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6781 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6841 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6901 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6961 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7021 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7081 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7141 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7201 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7261 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7321 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7381 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7441 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7501 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7561 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7621 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt agaaaaactc atcgagcatc
     7681 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     7741 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     7801 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     7861 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     7921 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     7981 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     8041 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     8101 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     8161 gttttcccag ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     8221 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     8281 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     8341 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     8401 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     8461 tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8521 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8581 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     8641 tataaaaata ggcgtatcac gaggcccttt cgtc
//
