LOCUS       sequence-96270-e        8611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains human dysferlin with exon 8 deleted. Deletion 
            maintains the open reading frame..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8611)
  TITLE     Human dysferlin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8611)
  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            "Unpublished"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-Exon8-del
FEATURES             Location/Qualifiers
     source          1..8611
                     /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      6426..6435
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6435..6451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6435..6451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6448..6470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6459..6475
                     /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(6483..6489,6490..6507,6508..6513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6528..6549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6666..6683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6837..7425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6917..6936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7596..8411)
                     /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(7596..8411)
                     /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     8319..8338
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8412..8516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8584..8602
                     /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 gtggtagact
     1021 ctcgttctct caggacagat gctctcctcg gggagttccg gatggacgtg ggcaccattt
     1081 acagagagcc ccggcacgcc tatctcagga agtggctgct gctctcagac cctgatgact
     1141 tctctgctgg ggccagaggc tacctgaaaa caagcctttg tgtgctgggg cctggggacg
     1201 aagcgcctct ggagagaaaa gacccctctg aagacaagga ggacattgaa agcaacctgc
     1261 tccggcccac aggcgtagcc ctgcgaggag cccacttctg cctgaaggtc ttccgggccg
     1321 aggacttgcc gcagatggac gatgccgtga tggacaacgt gaaacagatc tttggcttcg
     1381 agagtaacaa gaagaacttg gtggacccct ttgtggaggt cagctttgcg gggaaaatgc
     1441 tgtgcagcaa gatcttggag aagacggcca accctcagtg gaaccagaac atcacactgc
     1501 ctgccatgtt tccctccatg tgcgaaaaaa tgaggattcg tatcatagac tgggaccgcc
     1561 tgactcacaa tgacatcgtg gctaccacct acctgagtat gtcgaaaatc tctgcccctg
     1621 gaggagaaat agaagaggag cctgcaggtg ctgtcaagcc ttcgaaagcc tcagacttgg
     1681 atgactacct gggcttcctc cccacttttg ggccctgcta catcaacctc tatggcagtc
     1741 ccagagagtt cacaggcttc ccagacccct acacagagct caacacaggc aagggggaag
     1801 gtgtggctta tcgtggccgg cttctgctct ccctggagac caagctggtg gagcacagtg
     1861 aacagaaggt ggaggacctt cctgcggatg acatcctccg ggtggagaag taccttagga
     1921 ggcgcaagta ctccctgttt gcggccttct actcagccac catgctgcag gatgtggatg
     1981 atgccatcca gtttgaggtc agcatcggga actacgggaa caagttcgac atgacctgcc
     2041 tgccgctggc ctccaccact cagtacagcc gtgcagtctt tgacgggtgc cactactact
     2101 acctaccctg gggtaacgtg aaacctgtgg tggtgctgtc atcctactgg gaggacatca
     2161 gccatagaat cgagactcag aaccagctgc ttgggattgc tgaccggctg gaagctggcc
     2221 tggagcaggt ccacctggcc ctgaaggcgc agtgctccac ggaggacgtg gactcgctgg
     2281 tggctcagct gacggatgag ctcatcgcag gctgcagcca gcctctgggt gacatccatg
     2341 agacaccctc tgccacccac ctggaccagt acctgtacca gctgcgcacc catcacctga
     2401 gccaaatcac tgaggctgcc ctggccctga agctcggcca cagtgagctc cctgcagctc
     2461 tggagcaggc ggaggactgg ctcctgcgtc tgcgtgccct ggcagaggag ccccagaaca
     2521 gcctgccgga catcgtcatc tggatgctgc agggagacaa gcgtgtggca taccagcggg
     2581 tgcccgccca ccaagtcctc ttctcccggc ggggtgccaa ctactgtggc aagaattgtg
     2641 ggaagctaca gacaatcttt ctgaaatatc cgatggagaa ggtgcctggc gcccggatgc
     2701 cagtgcagat acgggtcaag ctgtggtttg ggctctctgt ggatgagaag gagttcaacc
     2761 agtttgctga ggggaagctg tctgtctttg ctgaaaccta tgagaacgag actaagttgg
     2821 cccttgttgg gaactggggc acaacgggcc tcacctaccc caagttttct gacgtcacgg
     2881 gcaagatcaa gctacccaag gacagcttcc gcccctcggc cggctggacc tgggctggag
     2941 attggttcgt gtgtccggag aagactctgc tccatgacat ggacgccggt cacctgagct
     3001 tcgtggaaga ggtgtttgag aaccagaccc ggcttcccgg aggccagtgg atctacatga
     3061 gtgacaacta caccgatgtg aacggggaga aggtgcttcc caaggatgac attgagtgcc
     3121 cactgggctg gaagtgggaa gatgaggaat ggtccacaga cctcaaccgg gctgtcgatg
     3181 agcaaggctg ggagtatagc atcaccatcc ccccggagcg gaagccgaag cactgggtcc
     3241 ctgctgagaa gatgtactac acacaccgac ggcggcgctg ggtgcgcctg cgcaggaggg
     3301 atctcagcca aatggaagca ctgaaaaggc acaggcaggc ggaggcggag ggcgagggct
     3361 gggagtacgc ctctcttttt ggctggaagt tccacctcga gtaccgcaag acagatgcct
     3421 tccgccgccg ccgctggcgc cgtcgcatgg agccactgga gaagacgggg cctgcagctg
     3481 tgtttgccct tgagggggcc ctgggcggcg tgatggatga caagagtgaa gattccatgt
     3541 ccgtctccac cttgagcttc ggtgtgaaca gacccacgat ttcctgcata ttcgactatg
     3601 ggaaccgcta ccatctacgc tgctacatgt accaggcccg ggacctggct gcgatggaca
     3661 aggactcttt ttctgatccc tatgccatcg tctccttcct gcaccagagc cagaagacgg
     3721 tggtggtgaa gaacaccctt aaccccacct gggaccagac gctcatcttc tacgagatcg
     3781 agatctttgg cgagccggcc acagttgctg agcaaccgcc cagcattgtg gtggagctgt
     3841 acgaccatga cacttatggt gcagacgagt ttatgggtcg ctgcatctgt caaccgagtc
     3901 tggaacggat gccacggctg gcctggttcc cactgacgag gggcagccag ccgtcggggg
     3961 agctgctggc ctcttttgag ctcatccaga gagagaagcc ggccatccac catattcctg
     4021 gttttgaggt gcaggagaca tcaaggatcc tggatgagtc tgaggacaca gacctgccct
     4081 acccaccacc ccagagggag gccaacatct acatggttcc tcagaacatc aagccagcgc
     4141 tccagcgtac cgccatcgag atcctggcat ggggcctgcg gaacatgaag agttaccagc
     4201 tggccaacat ctcctccccc agcctcgtgg tagagtgtgg gggccagacg gtgcagtcct
     4261 gtgtcatcag gaacctccgg aagaacccca actttgacat ctgcaccctc ttcatggaag
     4321 tgatgctgcc cagggaggag ctctactgcc cccccatcac cgtcaaggtc atcgataacc
     4381 gccagtttgg ccgccggcct gtggtgggcc agtgtaccat ccgctccctg gagagcttcc
     4441 tgtgtgaccc ctactcggcg gagagtccat ccccacaggg tggcccagac gatgtgagcc
     4501 tactcagtcc tggggaagac gtgctcatcg acattgatga caaggagccc ctcatcccca
     4561 tccaggagga agagttcatc gattggtgga gcaaattctt tgcctccata ggggagaggg
     4621 aaaagtgcgg ctcctacctg gagaaggatt ttgacaccct gaaggtctat gacacacagc
     4681 tggagaatgt ggaggccttt gagggcctgt ctgacttttg taacaccttc aagctgtacc
     4741 ggggcaagac gcaggaggag acagaagatc catctgtgat tggtgaattt aagggcctct
     4801 tcaaaattta tcccctccca gaagacccag ccatccccat gcccccaaga cagttccacc
     4861 agctggccgc ccagggaccc caggagtgct tggtccgtat ctacattgtc cgagcatttg
     4921 gcctgcagcc caaggacccc aatggaaagt gtgatcctta catcaagatc tccataggga
     4981 agaaatcagt gagtgaccag gataactaca tcccctgcac gctggagccc gtatttggaa
     5041 agatgttcga gctgacctgc actctgcctc tggagaagga cctaaagatc actctctatg
     5101 actatgacct cctctccaag gacgaaaaga tcggtgagac ggtcgtcgac ctggagaaca
     5161 ggctgctgtc caagtttggg gctcgctgtg gactcccaca gacctactgt gtctctggac
     5221 cgaaccagtg gcgggaccag ctccgcccct cccagctcct ccacctcttc tgccagcagc
     5281 atagagtcaa ggcacctgtg taccggacag accgtgtaat gtttcaggat aaagaatatt
     5341 ccattgaaga gatagaggct ggcaggatcc caaacccaca cctgggccca gtggaggagc
     5401 gtctggctct gcatgtgctt cagcagcagg gcctggtccc ggagcacgtg gagtcacggc
     5461 ccctctacag ccccctgcag ccagacatcg agcaggggaa gctgcagatg tgggtcgacc
     5521 tatttccgaa ggccctgggg cggcctggac ctcccttcaa catcacccca cggagagcca
     5581 gaaggttttt cctgcgttgt attatctgga ataccagaga tgtgatcctg gatgacctga
     5641 gcctcacggg ggagaagatg agcgacattt atgtgaaagg ttggatgatt ggctttgaag
     5701 aacacaagca aaagacagac gtgcattatc gttccctggg aggtgaaggc aacttcaact
     5761 ggaggttcat tttccccttc gactacctgc cagctgagca agtctgtacc attgccaaga
     5821 aggatgcctt ctggaggctg gacaagactg agagcaaaat cccagcacga gtggtgttcc
     5881 agatctggga caatgacaag ttctcctttg atgattttct gggctccctg cagctcgatc
     5941 tcaaccgcat gcccaagcca gccaagacag ccaagaagtg ctccttggac cagctggatg
     6001 atgctttcca cccagaatgg tttgtgtccc tttttgagca gaaaacagtg aagggctggt
     6061 ggccctgtgt agcagaagag ggtgagaaga aaatactggc gggcaagctg gaaatgacct
     6121 tggagattgt agcagagagt gagcatgagg agcggcctgc tggccagggc cgggatgagc
     6181 ccaacatgaa ccctaagctt gaggacccaa ggcgccccga cacctccttc ctgtggttta
     6241 cctccccata caagaccatg aagttcatcc tgtggcggcg tttccggtgg gccatcatcc
     6301 tcttcatcat cctcttcatc ctgctgctgt tcctggccat cttcatctac gccttcccga
     6361 actatgctgc catgaagctg gtgaagccct tcagctgatg atcaccatgg catatggcta
     6421 gcgcggccgc catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6481 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6541 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6601 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6661 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6721 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6781 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6841 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6901 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6961 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7021 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7081 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7141 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7201 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7261 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7321 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7381 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7441 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7501 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7561 atctaaagta tatatgagta aacttggtct gacagttaga aaaactcatc gagcatcaaa
     7621 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     7681 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     7741 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     7801 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     7861 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     7921 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     7981 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     8041 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     8101 ttcccaggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     8161 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     8221 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     8281 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     8341 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     8401 atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8461 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8521 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     8581 aaaaataggc gtatcacgag gccctttcgt c
//
