LOCUS       sequence-174247-        8957 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 8957)
  TITLE     Jain Foundation plasmids
REFERENCE   2  (bases 1 to 8957)
  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, 17
FEATURES             Location/Qualifiers
     source          1..8957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..35
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     27..44
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     28..44
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             831..842
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6430..6446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6430..6446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6443..6465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6454..6470
                     /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(6478..6484,6485..6502,6503..6508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6523..6544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6661..6678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6832..7420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6912..6931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7591..8406)
                     /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(7591..8406)
                     /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     8314..8333
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(8407..8511)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8579..8597
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8635..8657)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8757..8776
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg acgcgtatgc
       61 tgtgctgcct gctggtgagg gccagcaacc tccccagtgc gaagaaggac cggcgcagcg
      121 accctgtcgc aagcctgact ttccgagggg tgaagaagag aaccaaagtc atcaagaaca
      181 gcgtgaaccc tgtatggaat gagggatttg aatgggacct caagggcatc cccctggacc
      241 agggctctga gcttcatgtg gtggtcaaag accatgagac gatggggagg aacaggttcc
      301 tgggggaagc caaggtccca ctccgagagg tcctcgccac ccctagtctg tccgccagct
      361 tcaatgcccc cctgctggac accaagaagc agcccacagg ggcctcgctg gtcctgcagg
      421 tgtcctacac accgctgcct ggagctgtgc ccctgttccc gccccctact cctctggagc
      481 cctccccgac tctgcctgac ctggatgtag tggcaggcgg gggacagagc cgggccgaga
      541 cttggtccct gctcagtgac agcaccatgg acacgagata ctctggaaag aagtggccgg
      601 cccccacgga cacaggagga gaggaagaca cagaggacca gggactcact ggagatgagg
      661 cggagccatt cctggatcaa agcggaggcc cgggggctcc caccacccca aggaaactac
      721 cttcacgtcc tccgccccac taccccggga tcaaaagaaa gcgaagtgcg cctacatcta
      781 gaaagctgct gtcagacaaa ccgcaggatt tccagatcag ggtccaggtg atcgaggggc
      841 gccagctgcc gggggtgaac atcaagcctg tggtcaaggt taccgctgca gggcagacca
      901 agcggacgcg gatccacaag ggaaacagcc cactcttcaa tgagactctt ttcttcaact
      961 tgtttgactc tcctggggag ctgtttgatg agcccatctt tatcacggtg gtagactctc
     1021 gttctctcag gacagatgct ctcctcgggg agttccggat ggacgtgggc accatttaca
     1081 gagagccccg gcacgcctat ctcaggaagt ggctgctgct ctcagaccct gatgacttct
     1141 ctgctggggc cagaggctac ctgaaaacaa gcctttgtgt gctggggcct ggggacgaag
     1201 cgcctctgga gagaaaagac ccctctgaag acaaggagga cattgaaagc aacctgctcc
     1261 ggcccacagg cgtagccctg cgaggagccc acttctgcct gaaggtcttc cgggccgagg
     1321 acttgccgca gatggacgat gccgtgatgg acaacgtgaa acagatcttt ggcttcgaga
     1381 gtaacaagaa gaacttggtg gacccctttg tggaggtcag ctttgcgggg aaaatgctgt
     1441 gcagcaagat cttggagaag acggccaacc ctcagtggaa ccagaacatc acactgcctg
     1501 ccatgtttcc ctccatgtgc gaaaaaatga ggattcgtat catagactgg gaccgcctga
     1561 ctcacaatga catcgtggct accacctacc tgagtatgtc gaaaatctct gcccctggag
     1621 gagaaataga agaggagcct gcaggtgctg tcaagccttc gaaagcctca gacttggatg
     1681 actacctggg cttcctcccc acttttgggc cctgctacat caacctctat ggcagtccca
     1741 gagagttcac aggcttccca gacccctaca cagagctcaa cacaggcaag ggggaaggtg
     1801 tggcttatcg tggccggctt ctgctctccc tggagaccaa gctggtggag cacagtgaac
     1861 agaaggtgga ggaccttcct gcggatgaca tcctccgggt ggagaagtac cttaggaggc
     1921 gcaagtactc cctgtttgcg gccttctact cagccaccat gctgcaggat gtggatgacg
     1981 ccatccagtt tgaggtcagc atcgggaact acgggaacaa gttcgacatg acctgcctgc
     2041 cgctggcctc caccactcag tacagccgtg cagtctttga cgggtgccac tactactacc
     2101 taccctgggg taacgtgaaa cccgtggtgg tgctgtcatc ctactgggag gacatcagcc
     2161 atagaatcga gactcagaac cagctgcttg ggattgctga ccggctggaa gctggcctgg
     2221 agcaggtcca cctggccctg aaggcgcagt gctccacgga ggacgtggac tcgctggtgg
     2281 ctcagctgac ggatgagctc atcgcaggct gcagccagcc tctgggtgac atccatgaga
     2341 caccctctgc cacccacctg gaccagtacc tgtaccagct gcgcacccat cacctgagcc
     2401 aaatcactga ggctgccctg gccctgaagc tcggccacag tgagctccct gcagctctgg
     2461 agcaggcgga ggactggctc ctgcgtctgc gtgccctggc agaggagccc cagaacagcc
     2521 tgccggacat cgtcatctgg atgctgcagg gagacaagcg tgtggcatac cagcgggtgc
     2581 ctgcccacca agtcctcttc tcccggcggg gtgccaacta ctgtggcaag aattgtggga
     2641 agctacagac aatctttctg aaatatccga tggagaaggt gcctggcgcc cggatgccag
     2701 tgcagatacg ggtcaagctg tggtttgggc tctctgtgga tgagaaggag ttcaaccagt
     2761 ttgctgaggg gaagctgtct gtctttgctg aaacctatga gaacgagact aagttggccc
     2821 ttgttgggaa ctggggcaca acgggcctca cctaccccaa gttttctgac gtcacgggca
     2881 agatcaagct acccaaggac agcttccgcc cctcggccgg ctggacctgg gctggagatt
     2941 ggttcgtgtg cccggagaag actctgctcc atgacatgga cgccggtcac ctgagcttcg
     3001 tggaagaggt gtttgagaac cagacccggc ttcccggagg ccagtggatc tacatgagtg
     3061 acaactacac cgatgtgaac ggggagaagg tgcttcccaa ggatgacatt gagtgcccac
     3121 tgggctggaa gtgggaagat gaggaatggt ccacagacct caaccgggct gtcgatgagc
     3181 aaggctggga gtatagcatc accatccccc cggagcggaa gccgaagcac tgggtccctg
     3241 ctgagaagat gtactacaca caccgacggc ggcgctgggt gcgcctgcgc aggagggatc
     3301 tcagccaaat ggaagcactg aaaaggcaca ggcaggcgga ggcggagggc gagggctggg
     3361 agtacgcctc tctttttggc tggaagttcc acctcgagta ccgcaagaca gatgccttcc
     3421 gccgccgccg ctggcgccgt cgcatggagc cactggagaa gacggggcct gcagctgtgt
     3481 ttgcccttga gggggccctg ggcggcgtga tggatgacaa gagtgaagat tccatgtccg
     3541 tctccacctt gagcttcggt gtgaacagac ccacgatttc ctgcatattc gactatggga
     3601 accgctacca tctacgctgc tacatgtacc aggcccggga cctggctgcg atggacaagg
     3661 actctttttc tgatccctat gccatcgtct ccttcctgca ccagagccag aagacggtgg
     3721 tggtgaagaa cacccttaac cccacctggg accagacgct catcttctac gagatcgaga
     3781 tctttggcga gccggccaca gttgctgagc aaccgcccag cattgtggtg gagctgtacg
     3841 accatgacac ttatggtgca gacgagttta tgggtcgctg catctgtcaa ccgagtctgg
     3901 aacggatgcc acggctggcc tggttcccac tgacgagggg cagccagccg tcgggggagc
     3961 tgctggcctc ttttgagctc atccagagag agaagccggc catccaccat attcctggtt
     4021 ttgaggtgca ggagacatca aggatcctgg atgagtctga ggacacagac ctgccctacc
     4081 caccacccca gagggaggcc aacatctaca tggttcctca gaacatcaag ccagcgctcc
     4141 agcgtaccgc catcgagatc ctggcatggg gcctgcggaa catgaagagt taccagctgg
     4201 ccaacatctc ctcccccagc ctcgtggtag agtgtggggg ccagacggtg cagtcctgtg
     4261 tcatcaggaa cctccggaag aaccccaact ttgacatctg caccctcttc atggaagtga
     4321 tgctgcccag ggaggagctc tactgccccc ccatcaccgt caaggtcatc gataaccgcc
     4381 agtttggccg ccggcctgtg gtgggccagt gtaccatccg ctccctggag agcttcctgt
     4441 gtgaccccta ctcggcggag agtccatccc cacagggtgg cccagacgat gtgagcctac
     4501 tcagtcctgg ggaagacgtg ctcatcgaca ttgatgacaa ggagcccctc atccccatcc
     4561 aggaggaaga gttcatcgat tggtggagca aattctttgc ctccataggg gagagggaaa
     4621 agtgcggctc ctacctggag aaggattttg acaccctgaa ggtctatgac acacagctgg
     4681 agaatgtgga ggcctttgag ggcctgtctg acttttgtaa caccttcaag ctgtaccggg
     4741 gcaagacgca ggaggagaca gaagatccat ctgtgattgg tgaatttaag ggcctcttca
     4801 aaatttatcc cctcccagaa gacccagcca tccccatgcc cccaagacag ttccaccagc
     4861 tggccgccca gggaccccag gagtgcttgg tccgtatcta cattgtccga gcatttggcc
     4921 tgcagcccaa ggaccccaat ggaaagtgtg atccttacat caagatctcc atagggaaga
     4981 aatcagtgag tgaccaggat aactacatcc cctgcacgct ggagcccgta tttggaaaga
     5041 tgttcgagct gacctgcact ctgcctctgg agaaggacct aaagatcact ctctatgact
     5101 atgacctcct ctccaaggac gaaaagatcg gtgagacggt cgtcgacctg gagaacaggc
     5161 tgctgtccaa gtttggggct cgctgtggac tcccacagac ctactgtgtc tctggaccga
     5221 accagtggcg ggaccagctc cgcccctccc agctcctcca cctcttctgc cagcagcata
     5281 gagtcaaggc acctgtgtac cggacagacc gtgtaatgtt tcaggataaa gaatattcca
     5341 ttgaagagat agaggctggc aggatcccaa acccacacct gggcccagtg gaggagcgtc
     5401 tggctctgca tgtgcttcag cagcagggcc tggtcccgga gcacgtggag tcacggcccc
     5461 tctacagccc cctgcagcca gacatcgagc aggggaagct gcagatgtgg gtcgacctat
     5521 ttccgaaggc cctggggcgg cctggacctc ccttcaacat caccccacgg agagccagaa
     5581 ggtttttcct gcgttgtatt atctggaata ccagagatgt gatcctggat gacctgagcc
     5641 tcacggggga gaagatgagc gacatttatg tgaaaggttg gatgattggc tttgaagaac
     5701 acaagcaaaa gacagacgtg cattatcgtt ccctgggagg tgaaggcaac ttcaactgga
     5761 ggttcatttt ccccttcgac tacctgccag ctgagcaagt ctgtaccatt gccaagaagg
     5821 atgccttctg gaggctggac aagactgaga gcaaaatccc agcacgagtg gtgttccaga
     5881 tctgggacaa tgacaagttc tcctttgatg attttctggg ctccctgcag ctcgatctca
     5941 accgcatgcc caagccagcc aagacagcca agaagtgctc cttggaccag ctggatgatg
     6001 ctttccaccc agaatggttt gtgtcccttt ttgagcagaa aacagtgaag ggctggtggc
     6061 cctgtgtagc agaagagggt gagaagaaaa tactggcggg caagctggaa atgaccttgg
     6121 agattgtagc agagagtgag catgaggagc ggcctgctgg ccagggccgg gatgagccca
     6181 acatgaaccc taagcttgag gacccaaggc gccccgacac ctccttcctg tggtttacct
     6241 ccccatacaa gaccatgaag ttcatcctgt ggcggcgttt ccggtgggcc atcatcctct
     6301 tcatcatcct cttcatcctg ctgctgttcc tggccatctt catctacgcc ttcccgaact
     6361 atgctgccat gaagctggtg aagcccttca gctgaatccc aatggcgcgc cgagcttggc
     6421 tcgagcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6601 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6661 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7261 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7381 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7501 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7561 aagtatatat gagtaaactt ggtctgacag ttagaaaaac tcatcgagca tcaaatgaaa
     7621 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     7681 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     7741 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     7801 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     7861 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     7921 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     7981 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     8041 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     8101 agggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     8161 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     8221 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     8281 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     8341 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     8401 gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8461 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8521 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8581 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8641 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8701 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     8761 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     8821 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     8881 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     8941 gcgattaagt tgggtaa
//
