LOCUS       sequence-193244-        4127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of shRNA targeting SRF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4127)
  AUTHORS   Mughal W, Nguyen L, Pustylnik S, da Silva Rosa SC, Piotrowski S, 
            Chapman D, Du M, Alli NS, Grigull J, Halayko AJ, Aliani M, Topham 
            MK, Epand RM, Hatch GM, Pereira TJ, Kereliuk S, McDermott JC, 
            Rampitsch C, Dolinsky VW, Gordon JW
  TITLE     A conserved MADS-box phosphorylation motif regulates differentiation
            and mitochondrial function in skeletal, cardiac, and smooth muscle 
            cells.
  JOURNAL   Cell Death Dis. 2015 Oct 29;6:e1944. doi: 10.1038/cddis.2015.306.
  PUBMED    26512955
REFERENCE   2  (bases 1 to 4127)
  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 
            Dis."; date: "2015-10-29"; volume: "6:e1944. doi"; pages: " 
            10.1038/cddis.2015.306"
COMMENT     Sequence Label: sh-SRF
FEATURES             Location/Qualifiers
     source          1..4127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     760..777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    894..915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(930..935,936..953,954..960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    968..984
                     /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)."
     primer_bind     973..995
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     992..1008
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     992..1008
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(1093..1183)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(1153..1172)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(1190..1207)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1190..1206)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1199..1221)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1415..1434)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1534..1556
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1594..1612)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    1771..1852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1796..1815
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2082..2681)
                     /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(2165..2185)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2662..2681
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2778..2974)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(2786..2996)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2792..2927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(3114..3182,3183..3974)
                     /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     complement(3332..3351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      781 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      841 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      901 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      961 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     1021 aagcttttcc aaaaaactgc agcccatgat caccatctct tgaatggtga tcatgggctg
     1081 cagcggatcc gagtggtctc atacagaact tataagattc ccaaatccaa agacatttca
     1141 cgtttatggt gatttcccag aacacatagc gacatgcaaa tatgaattca ctggccgtcg
     1201 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     1261 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1321 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     1381 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     1441 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     1501 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     1561 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     1621 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1681 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1741 aataaccctg ataaatgctt caataatatt gcagtgaaaa aaatgcttta tttgtgaaat
     1801 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1861 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1921 gtaaaacctc tacaaatgtg gtatggctga ttatgatcct ctagagtcgg tgggcctcgg
     1981 gggcgggtgc ggggtcggcg gggccgcccc gggtggcttc ggtcggagcc atggggtcgt
     2041 gcgctccttt cggtcgggcg ctgcgggtcg tggggcgggc gtcaggcacc gggcttgcgg
     2101 gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg
     2161 agccgctcgt agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg
     2221 gcgcgctcgg ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg
     2281 tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc
     2341 gccaggaggc cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg
     2401 cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc
     2461 gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg
     2521 aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccgggtc gacggtgtgg
     2581 cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggcccg ggggacgtcg
     2641 tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca tggaaggtcg tctccttgtg
     2701 aggggtcagg ggcgtgggtc aggggatggt ggcggcaccg gtcgtggcgg ccgacctgca
     2761 ggcatgcaag ctagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     2821 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     2881 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     2941 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     3001 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     3061 ctgcctgctg gggagcctgg ggactttcca accaatattg aaaaaggaag agtatgagta
     3121 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3181 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3241 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3301 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3361 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3421 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3481 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3541 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3601 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3661 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3721 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3781 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3841 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3901 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3961 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4021 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4081 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaa
//
