LOCUS       sequence-174249-        8915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  research of alternative isoforms of the protein dysferlin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8915)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8915)
  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"
COMMENT     Sequence Label: pUC57 DYSF, alt exon 1a, 5a
FEATURES             Location/Qualifiers
     source          1..8915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..36
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     28..45
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     29..45
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             832..843
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6389..6405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6389..6405)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6402..6424)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6413..6429
                     /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(6437..6443,6444..6461,6462..6467))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6482..6503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6620..6637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6791..7379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6871..6890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7550..8365)
                     /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(7550..8365)
                     /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     8273..8292
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8366..8470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8538..8556
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8594..8616)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8716..8735
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatg
       61 ctgtgctgcc tgctggtgag ggccagcaac ctccccagtg cgaagaagga ccggcgcagc
      121 gaccctgtcg caagcctgac tttccgaggg gtgaagaaga gaaccaaagt catcaagaac
      181 agcgtgaacc ctgtatggaa tgagggattt gaatgggacc tcaagggcat ccccctggac
      241 cagggctctg agcttcatgt ggtggtcaaa gaccatgaga cgatggggag gaacaggttc
      301 ctgggggaag ccaaggtccc actccgagag gtcctcgcca cccctagtct gtccgccagc
      361 ttcaatgccc ccctgctgga caccaagaag cagcccacag gggcctcgct ggtcctgcag
      421 gtgtcctaca caccgctgcc tggagctgtg cccctgttcc cgccccctac tcctctggag
      481 ccctccccga ctctgcctga cctggatgta gtggcaggcg ggggacagag ccgggccgag
      541 acttggtccc tgctcagtga cagcaccatg gacacgagat actctggaaa gaagtggccg
      601 gcccccacgg acacaggagg agaggaagac acagaggacc agggactcac tggagatgag
      661 gcggagccat tcctggatca aagcggaggc ccgggggctc ccaccacccc aaggaaacta
      721 ccttcacgtc ctccgcccca ctaccccggg atcaaaagaa agcgaagtgc gcctacatct
      781 agaaagctgc tgtcagacaa accgcaggat ttccagatca gggtccaggt gatcgagggg
      841 cgccagctgc cgggggtgaa catcaagcct gtggtcaagg ttaccgctgc agggcagacc
      901 aagcggacgc ggatccacaa gggaaacagc ccactcttca atgagactct tttcttcaac
      961 ttgtttgact ctcctgggga gctgtttgat gagcccatct ttatcacggt ggtagactct
     1021 cgttctctca ggacagatgc tctcctcggg gagttccgga tggacgtggg caccatttac
     1081 agagagcccc ggcacgccta tctcaggaag tggctgctgc tctcagaccc tgatgacttc
     1141 tctgctgggg ccagaggcta cctgaaaaca agcctttgtg tgctggggcc tggggacgaa
     1201 gcgcctctgg agagaaaaga cccctctgaa gacaaggagg acattgaaag caacctgctc
     1261 cggcccacag gcgtagccct gcgaggagcc cacttctgcc tgaaggtctt ccgggccgag
     1321 gacttgccgc agatggacga tgccgtgatg gacaacgtga aacagatctt tggcttcgag
     1381 agtaacaaga agaacttggt ggaccccttt gtggaggtca gctttgcggg gaaaatgctg
     1441 tgcagcaaga tcttggagaa gacggccaac cctcagtgga accagaacat cacactgcct
     1501 gccatgtttc cctccatgtg cgaaaaaatg aggattcgta tcatagactg ggaccgcctg
     1561 actcacaatg acatcgtggc taccacctac ctgagtatgt cgaaaatctc tgcccctgga
     1621 ggagaaatag aagtggatga ctacctgggc ttcctcccca cttttgggcc ctgctacatc
     1681 aacctctatg gcagtcccag agagttcaca ggcttcccag acccctacac agagctcaac
     1741 acaggcaagg gggaaggtgt ggcttatcgt ggccggcttc tgctctccct ggagaccaag
     1801 ctggtggagc acagtgaaca gaaggtggag gaccttcctg cggatgacat cctccgggtg
     1861 gagaagtacc ttaggaggcg caagtactcc ctgtttgcgg ccttctactc agccaccatg
     1921 ctgcaggatg tggatgacgc catccagttt gaggtcagca tcgggaacta cgggaacaag
     1981 ttcgacatga cctgcctgcc gctggcctcc accactcagt acagccgtgc agtctttgac
     2041 gggtgccact actactacct accctggggt aacgtgaaac ccgtggtggt gctgtcatcc
     2101 tactgggagg acatcagcca tagaatcgag actcagaacc agctgcttgg gattgctgac
     2161 cggctggaag ctggcctgga gcaggtccac ctggccctga aggcgcagtg ctccacggag
     2221 gacgtggact cgctggtggc tcagctgacg gatgagctca tcgcaggctg cagccagcct
     2281 ctgggtgaca tccatgagac accctctgcc acccacctgg accagtacct gtaccagctg
     2341 cgcacccatc acctgagcca aatcactgag gctgccctgg ccctgaagct cggccacagt
     2401 gagctccctg cagctctgga gcaggcggag gactggctcc tgcgtctgcg tgccctggca
     2461 gaggagcccc agaacagcct gccggacatc gtcatctgga tgctgcaggg agacaagcgt
     2521 gtggcatacc agcgggtgcc tgcccaccaa gtcctcttct cccggcgggg tgccaactac
     2581 tgtggcaaga attgtgggaa gctacagaca atctttctga aatatccgat ggagaaggtg
     2641 cctggcgccc ggatgccagt gcagatacgg gtcaagctgt ggtttgggct ctctgtggat
     2701 gagaaggagt tcaaccagtt tgctgagggg aagctgtctg tctttgctga aacctatgag
     2761 aacgagacta agttggccct tgttgggaac tggggcacaa cgggcctcac ctaccccaag
     2821 ttttctgacg tcacgggcaa gatcaagcta cccaaggaca gcttccgccc ctcggccggc
     2881 tggacctggg ctggagattg gttcgtgtgc ccggagaaga ctctgctcca tgacatggac
     2941 gccggtcacc tgagcttcgt ggaagaggtg tttgagaacc agacccggct tcccggaggc
     3001 cagtggatct acatgagtga caactacacc gatgtgaacg gggagaaggt gcttcccaag
     3061 gatgacattg agtgcccact gggctggaag tgggaagatg aggaatggtc cacagacctc
     3121 aaccgggctg tcgatgagca aggctgggag tatagcatca ccatcccccc ggagcggaag
     3181 ccgaagcact gggtccctgc tgagaagatg tactacacac accgacggcg gcgctgggtg
     3241 cgcctgcgca ggagggatct cagccaaatg gaagcactga aaaggcacag gcaggcggag
     3301 gcggagggcg agggctggga gtacgcctct ctttttggct ggaagttcca cctcgagtac
     3361 cgcaagacag atgccttccg ccgccgccgc tggcgccgtc gcatggagcc actggagaag
     3421 acggggcctg cagctgtgtt tgcccttgag ggggccctgg gcggcgtgat ggatgacaag
     3481 agtgaagatt ccatgtccgt ctccaccttg agcttcggtg tgaacagacc cacgatttcc
     3541 tgcatattcg actatgggaa ccgctaccat ctacgctgct acatgtacca ggcccgggac
     3601 ctggctgcga tggacaagga ctctttttct gatccctatg ccatcgtctc cttcctgcac
     3661 cagagccaga agacggtggt ggtgaagaac acccttaacc ccacctggga ccagacgctc
     3721 atcttctacg agatcgagat ctttggcgag ccggccacag ttgctgagca accgcccagc
     3781 attgtggtgg agctgtacga ccatgacact tatggtgcag acgagtttat gggtcgctgc
     3841 atctgtcaac cgagtctgga acggatgcca cggctggcct ggttcccact gacgaggggc
     3901 agccagccgt cgggggagct gctggcctct tttgagctca tccagagaga gaagccggcc
     3961 atccaccata ttcctggttt tgaggtgcag gagacatcaa ggatcctgga tgagtctgag
     4021 gacacagacc tgccctaccc accaccccag agggaggcca acatctacat ggttcctcag
     4081 aacatcaagc cagcgctcca gcgtaccgcc atcgagatcc tggcatgggg cctgcggaac
     4141 atgaagagtt accagctggc caacatctcc tcccccagcc tcgtggtaga gtgtgggggc
     4201 cagacggtgc agtcctgtgt catcaggaac ctccggaaga accccaactt tgacatctgc
     4261 accctcttca tggaagtgat gctgcccagg gaggagctct actgcccccc catcaccgtc
     4321 aaggtcatcg ataaccgcca gtttggccgc cggcctgtgg tgggccagtg taccatccgc
     4381 tccctggaga gcttcctgtg tgacccctac tcggcggaga gtccatcccc acagggtggc
     4441 ccagacgatg tgagcctact cagtcctggg gaagacgtgc tcatcgacat tgatgacaag
     4501 gagcccctca tccccatcca ggaggaagag ttcatcgatt ggtggagcaa attctttgcc
     4561 tccatagggg agagggaaaa gtgcggctcc tacctggaga aggattttga caccctgaag
     4621 gtctatgaca cacagctgga gaatgtggag gcctttgagg gcctgtctga cttttgtaac
     4681 accttcaagc tgtaccgggg caagacgcag gaggagacag aagatccatc tgtgattggt
     4741 gaatttaagg gcctcttcaa aatttatccc ctcccagaag acccagccat ccccatgccc
     4801 ccaagacagt tccaccagct ggccgcccag ggaccccagg agtgcttggt ccgtatctac
     4861 attgtccgag catttggcct gcagcccaag gaccccaatg gaaagtgtga tccttacatc
     4921 aagatctcca tagggaagaa atcagtgagt gaccaggata actacatccc ctgcacgctg
     4981 gagcccgtat ttggaaagat gttcgagctg acctgcactc tgcctctgga gaaggaccta
     5041 aagatcactc tctatgacta tgacctcctc tccaaggacg aaaagatcgg tgagacggtc
     5101 gtcgacctgg agaacaggct gctgtccaag tttggggctc gctgtggact cccacagacc
     5161 tactgtgtct ctggaccgaa ccagtggcgg gaccagctcc gcccctccca gctcctccac
     5221 ctcttctgcc agcagcatag agtcaaggca cctgtgtacc ggacagaccg tgtaatgttt
     5281 caggataaag aatattccat tgaagagata gaggctggca ggatcccaaa cccacacctg
     5341 ggcccagtgg aggagcgtct ggctctgcat gtgcttcagc agcagggcct ggtcccggag
     5401 cacgtggagt cacggcccct ctacagcccc ctgcagccag acatcgagca ggggaagctg
     5461 cagatgtggg tcgacctatt tccgaaggcc ctggggcggc ctggacctcc cttcaacatc
     5521 accccacgga gagccagaag gtttttcctg cgttgtatta tctggaatac cagagatgtg
     5581 atcctggatg acctgagcct cacgggggag aagatgagcg acatttatgt gaaaggttgg
     5641 atgattggct ttgaagaaca caagcaaaag acagacgtgc attatcgttc cctgggaggt
     5701 gaaggcaact tcaactggag gttcattttc cccttcgact acctgccagc tgagcaagtc
     5761 tgtaccattg ccaagaagga tgccttctgg aggctggaca agactgagag caaaatccca
     5821 gcacgagtgg tgttccagat ctgggacaat gacaagttct cctttgatga ttttctgggc
     5881 tccctgcagc tcgatctcaa ccgcatgccc aagccagcca agacagccaa gaagtgctcc
     5941 ttggaccagc tggatgatgc tttccaccca gaatggtttg tgtccctttt tgagcagaaa
     6001 acagtgaagg gctggtggcc ctgtgtagca gaagagggtg agaagaaaat actggcgggc
     6061 aagctggaaa tgaccttgga gattgtagca gagagtgagc atgaggagcg gcctgctggc
     6121 cagggccggg atgagcccaa catgaaccct aagcttgagg acccaaggcg ccccgacacc
     6181 tccttcctgt ggtttacctc cccatacaag accatgaagt tcatcctgtg gcggcgtttc
     6241 cggtgggcca tcatcctctt catcatcctc ttcatcctgc tgctgttcct ggccatcttc
     6301 atctacgcct tcccgaacta tgctgccatg aagctggtga agcccttcag ctgaatccca
     6361 atggcgcgcc gagcttggct cgagcatggt catagctgtt tcctgtgtga aattgttatc
     6421 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6481 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6541 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6601 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6661 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6721 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6781 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6841 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6901 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6961 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7021 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7081 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7141 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7201 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7261 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7321 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7381 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7441 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7501 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt tagaaaaact
     7561 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     7621 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     7681 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     7741 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     7801 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     7861 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     7921 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     7981 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     8041 cctggaatgc tgttttccca gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     8101 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     8161 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     8221 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     8281 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     8341 acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga agcatttatc
     8401 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8461 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     8521 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     8581 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     8641 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     8701 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     8761 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     8821 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     8881 aagggggatg tgctgcaagg cgattaagtt gggta
//
