LOCUS       sequence-380204-        4179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh6 gene with CDS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4179)
  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 4179)
  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-mMyh6CDS
FEATURES             Location/Qualifiers
     source          1..4179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     414..433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(652..756)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(834..1289)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(971..992)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1183..1202
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1415..1437
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1429..1446
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1430..1446
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1453..1472
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1453..1471
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1502..1511
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1506..1515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2773..2791)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2774..2791)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2809..2825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2809..2825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2822..2844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2833..2849
                     /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(2857..2863,2864..2881,2882..2887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2902..2923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3040..3057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3211..3799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3291..3310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3970..4179,1..582,583..651))
                     /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 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
       61 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      121 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      181 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      241 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      301 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      361 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      421 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      481 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      541 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      601 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      661 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      721 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      781 tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
      841 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
      901 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
      961 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1021 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1081 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1141 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1201 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     1261 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     1321 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1381 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1441 gccagtgaat tgtaatacga ctcactatag ggcgaattgg gcccgacgtc gcatgctccc
     1501 ggccgccatg gcggccgcgg gaattcgatt ggatgccata cagaggacgg aggagctgga
     1561 ggaagccaag aagaagctgg ctcagaggct gcaggatgca gaggaggcag tggaggccgt
     1621 caacgccaag tgttcctccc tggagaagac caagcacagg ctgcagaatg agatcgagga
     1681 cctgatggtg gacgtggagc gctccaatgc cgccgccgca gccctggaca agaagcagag
     1741 gaactttgac aagatcctgg ctgagtggaa gcagaagtat gaggagtcgc agtcagagct
     1801 ggagtcttcc cagaaggagg cgcgctccct gagcacagag ctcttcaagc tcaagaacgc
     1861 ctatgaggag tctctggagc acctggagac cttcaagcgg gagaacaaga acctccagga
     1921 ggagatctca gacctgactg aacagctggg agaagggggg aaaaacgtgc acgagctgga
     1981 gaagatccgc aaacagctgg aggtggagaa gctggagctg cagtcagccc tggaggaggc
     2041 tgaggcctcc ctggagcacg aggagggcaa gatcctccgt gcccagctgg agttcaacca
     2101 gatcaaggca gagatcgaaa ggaagctggc agagaaggat gaggagatgg agcaggccaa
     2161 gcgcaaccac ctgcggatgg tggactccct gcagacctcc ctggatgcgg agacacgcag
     2221 ccgcaatgag gccctgcggg tgaagaagaa gatggagggc gacctcaacg agatggagat
     2281 ccagctcagc caggccaata gaatagcctc agaggcacag aaacacctga agaattctca
     2341 agctcacttg aaggacaccc agctccagct ggatgatgct gtccatgcca atgacgacct
     2401 gaaggagaac atcgccatcg tggaacggcg caacaacctg ctgcaggcgg agctggagga
     2461 gctgcgggct gtggtggagc agacggagcg gtctcggaag ctggcagagc aggagctgat
     2521 tgagaccagc gagcgggtgc agctgctgca ctcgcagaac accagcctca tcaaccagaa
     2581 gaagaagatg gagtcagacc tgacccaact ccagacagaa gtagaggagg cagtgcagga
     2641 gtgtaggaac gcagaggaga aggccaagaa ggccatcaca gatgccgcaa tgaatcacta
     2701 gtgaattcgc ggccgcctgc aggtcgacca tatgggagag ctcccaacgc gttggatgca
     2761 tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt catagctgtt
     2821 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2881 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2941 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3001 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3061 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3121 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3181 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3241 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3301 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3361 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3421 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3481 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3541 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3601 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3661 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3721 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3781 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3841 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3901 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3961 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4021 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4081 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4141 agcaataaac cagccagccg gaagggccga gcgcagaag
//
