LOCUS       sequence-380192-        4077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh3 gene with CDS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4077)
  AUTHORS   Tanji M, Wada K, Sakamoto K, Ono Y, Inui M
  TITLE     Digoxigenin-labeled RNA probes for untranslated regions enable the 
            isoform-specific gene expression analysis of myosin heavy chains in 
            whole-mount in situ hybridization.
  JOURNAL   Dev Growth Differ. 2022 Dec 14. doi: 10.1111/dgd.12832.
  PUBMED    36517457
REFERENCE   2  (bases 1 to 4077)
  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: "Dev Growth 
            Differ. 2022 Dec 14. doi: 10.1111/dgd.12832."
COMMENT     Sequence Label: pGEMT-mMyh3CDS
FEATURES             Location/Qualifiers
     source          1..4077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     285..304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(523..627)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(705..1160)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(842..863)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1054..1073
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1286..1308
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1300..1317
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1301..1317
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1324..1343
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1324..1342
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1373..1382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1377..1386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2542..2560)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2543..2560)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2578..2594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2578..2594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2591..2613)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2602..2618
                     /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(2626..2632,2633..2650,2651..2656))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2671..2692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2809..2826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2980..3568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3060..3079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3739..4077,1..453,454..522))
                     /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"
ORIGIN
        1 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
       61 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      121 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      181 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      241 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
      301 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
      361 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
      421 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
      481 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
      541 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
      601 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt
      661 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
      721 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
      781 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
      841 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
      901 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
      961 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     1021 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     1081 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     1141 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1201 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1261 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     1321 ttgtaatacg actcactata gggcgaattg ggcccgacgt cgcatgctcc cggccgccat
     1381 ggcggccgcg ggaattcgat tcgttttgga cattgcgggt tttgaaattt ttgagtataa
     1441 cagcctggag cagctgtgca tcaacttcac caacgagaaa ctgcaacagt ttttcaacca
     1501 ccacatgttc gtgctggaac aggaggaata caagaaggag ggcatcgagt ggaccttcat
     1561 cgacttcggg atggacctgg cagcctgcat cgagctcatc gagaagccga tgggcatctt
     1621 ctccatcctg gaagaggagt gcatgttccc caaggcgaca gacacctcct tcaagaacaa
     1681 gctgtatgac cagcacctgg gaaagtccaa caacttccag aaacctaagg tggtcaaagg
     1741 caaggctgaa gcccacttct ccctggtgca ctatgcgggc accgtggact acagtgtctc
     1801 aggctggctg gagaagaaca aggaccccct gaatgagacc gtggtgggac tgtaccagaa
     1861 gtcctccaac agactcctgg cacatctcta tgccaccttc gctacaacag atgcggacgg
     1921 tgggaagaag aaagtcgcta agaagaaagg ctcttccttc caaaccgtct ctgcactgtt
     1981 cagggaaaac ttgaacaagc tgatgtccaa tttaagaacc acacacccac actttgtacg
     2041 ctgtataata cccaacgaaa ccaagacccc aggggccatg gaacacagcc ttgtcctgca
     2101 ccagctcagg tgtaacgggg tgctggaagg catccggatc tgcaggaagg gcttccccaa
     2161 caggatcctg tatggagact ttaaacagag ataccgagtg ctgaacgcca gtgccatccc
     2221 agaaggccag ttcattgaca gcaagaaagc ttgcgagaaa ctcttggctt ccattgacat
     2281 cgaccacaca cagtacaaat ttggacacac caaggtgttc tttaaggctg gcttgctagg
     2341 aaccctggaa gagatgcggg atgagcgttt ggccaaactg atcactcgga cacaagccgt
     2401 gtgcagaggg tttctcatgc gtgtggaatt ccagaagatg atgcagagga gggagtccat
     2461 aaatcactag tgaattcgcg gccgcctgca ggtcgaccat atgggagagc tcccaacgcg
     2521 ttggatgcat agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc
     2581 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2641 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2701 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2761 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2821 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2881 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2941 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3001 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3061 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3121 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3181 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3241 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3301 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3361 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3421 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3481 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3541 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3601 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3661 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3721 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3781 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3841 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3901 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3961 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4021 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctc
//
