LOCUS       sequence-380184-        3567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh1 gene with CDS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3567)
  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 3567)
  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-mMyh1CDS
FEATURES             Location/Qualifiers
     source          1..3567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     574..593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(812..916)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(994..1449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1131..1152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1343..1362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1575..1597
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1589..1606
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1590..1606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1613..1632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1613..1631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1662..1671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1666..1675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2321..2339)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2322..2339)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2357..2373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2357..2373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2370..2392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2381..2397
                     /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(2405..2411,2412..2429,2430..2435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2450..2471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2588..2605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2759..3347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2839..2858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3518..3567,1..742,743..811))
                     /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 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
       61 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
      301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
      361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
      421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
      481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
      541 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
      601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
      661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
      721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
      781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
      841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg
      961 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa
     1021 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     1081 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     1141 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     1201 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     1261 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     1321 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     1381 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     1441 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     1501 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     1561 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     1621 ctcactatag ggcgaattgg gcccgacgtc gcatgctccc ggccgccatg gcggccgcgg
     1681 gaattcgatt ggaggagctg aagaaggagc aggataccag cgcccacctg gagcggatga
     1741 agaagaacct ggagcagacg gtgaaggacc tgcagcaccg tctggatgag gctgagcagc
     1801 tggcgctgaa gggcggcaag aagcagatcc agaaactgga ggccagggtc cgtgaacttg
     1861 aaggagaagt ggaaaatgaa cagaagcgca acgtggaagc tatcaagggt ctgcgcaaac
     1921 atgagagaag agtgaaagaa ctcacttatc aaactgagga agaccgcaag aatgttctca
     1981 ggctccagga ccttgtggac aaactgcaat caaaggtcaa ggcctacaag agacaagctg
     2041 aggaagcgga ggaacaatcc aatgtcaacc tggccaaatt ccgcaagatc cagcacgagc
     2101 tggaggaagc cgaggagcgg gctgacatcg cggagtccca ggtcaacaag ctgcgggtga
     2161 agagccggga ggttcacacc aaaatcataa gcgaagagtg attgatccaa gtgcaggaaa
     2221 gtgaccaaag agatgagcaa aatcactagt gaattcgcgg ccgcctgcag gtcgaccata
     2281 tgggagagct cccaacgcgt tggatgcata gcttgagtat tctatagtgt cacctaaata
     2341 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2401 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2461 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2521 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2581 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2641 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2941 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3181 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3541 cacctatctc agcgatctgt ctatttc
//
