LOCUS       sequence-193079-        4126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of shRNA targeting MYOCD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4126)
  AUTHORS   Mughal W, Martens M, Field J, Chapman D, Huang J, Rattan S, Hai Y, 
            Cheung KG, Kereliuk S, West AR, Cole LK, Hatch GM, Diehl-Jones W, 
            Keijzer R, Dolinsky VW, Dixon IM, Parmacek MS, Gordon JW
  TITLE     Myocardin regulates mitochondrial calcium homeostasis and prevents 
            permeability transition.
  JOURNAL   Cell Death Differ. 2018 Mar 6. pii: 10.1038/s41418-018-0073-z. doi: 
            10.1038/s41418-018-0073-z.
  PUBMED    29511336
REFERENCE   2  (bases 1 to 4126)
  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: "Cell Death 
            Differ. 2018 Mar 6. pii: 10.1038/s41418-018-0073-z. doi: 
            10.1038/s41418-018-0073-z."
COMMENT     Sequence Label: sh-Myocardin
FEATURES             Location/Qualifiers
     source          1..4126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..28
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     20..37
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     21..37
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        44..134
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     55..74
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(218..234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(218..234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(231..253)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    242..258
                     /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(266..272,273..290,291..296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    311..332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(449..466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(620..1208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(700..719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1379..2170,2171..2239))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2002..2021
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        2357..2567
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2379..2575
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2426..2561
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2672..3271
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2672..2691)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3168..3188
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3501..3582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3538..3557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3741..3759
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3797..3819)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3919..3938
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcatatttg catgtcgcta
       61 tgtgttctgg gaaatcacca taaacgtgaa atgtctttgg atttgggaat cttataagtt
      121 ctgtatgaga ccactcggat ccgttccgat cagtcttaca gttcaagaga ctgtaagact
      181 gatcggaact tttttggaaa agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      241 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      301 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      361 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      421 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      481 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      541 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      601 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      661 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      721 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      781 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      841 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      901 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      961 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1021 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1081 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1141 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1201 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1261 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1321 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1381 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1441 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1501 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1561 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1621 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1681 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1741 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1801 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1861 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1921 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1981 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2041 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2101 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2161 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2221 ggaaatgttg aatactcata ctcttccttt ttcaatattg gttggaaagt ccccaggctc
     2281 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2401 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2461 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2521 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2581 agcttgcatg cctgcaggtc ggccgccacg accggtgccg ccaccatccc ctgacccacg
     2641 cccctgaccc ctcacaagga gacgaccttc catgaccgag tacaagccca cggtgcgcct
     2701 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     2761 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     2821 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     2881 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     2941 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     3001 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     3061 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     3121 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     3181 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     3241 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     3301 cgaaaggagc gcacgacccc atggctccga ccgaagccac ccggggcggc cccgccgacc
     3361 ccgcacccgc ccccgaggcc caccgactct agaggatcat aatcagccat accacatttg
     3421 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3481 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3541 atagcatcac aaatttcaca aataaagcat ttttttcact gcaatattat tgaagcattt
     3601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3721 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     3781 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3841 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3901 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     3961 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     4021 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     4081 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagg
//
