LOCUS       sequence-380202-        4016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh4 gene with CDS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4016)
  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 4016)
  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-mMyh4CDS
FEATURES             Location/Qualifiers
     source          1..4016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      578..1166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1067..1086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1320..1337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1454..1475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1490..1495,1496..1513,1514..1520)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1528..1544
                     /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     1533..1555
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1552..1568
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1552..1568
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1586..1604
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1586..1603
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      2699..2708
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2703..2712
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2742..2761)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2743..2761)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2768..2785)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2768..2784)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2777..2799)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2925..3380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3012..3031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3222..3243
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3458..3562
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3563..3631,3632..4016,1..407)
                     /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(3781..3800)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
       61 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
      121 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
      181 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
      241 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
      301 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
      361 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
      421 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
      481 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
      541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      721 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      961 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1261 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1321 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     1381 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     1441 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     1501 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     1561 gctatgacca tgattacgcc aagctattta ggtgacacta tagaatactc aagctatgca
     1621 tccaacgcgt tgggagctct cccatatggt cgacctgcag gcggccgcga attcactagt
     1681 gatttgacga ccttgagctg acactggcca aggttgagaa ggagaagcac gccacggaga
     1741 acaaggtgaa aaacctgaca gaagagatgg caggcctgga tgaaaatatc gccaagctga
     1801 ccaaggagaa gaaggccctc caggaggccc accagcagac cctggatgac ctgcaggcag
     1861 aggaggacaa agtcaacacc ctgaccaaag ccaaaaccaa gctggaacag caagtggatg
     1921 accttgaagg atcactggaa caggaaaaga agctgaggat ggacttagaa agagccaaga
     1981 ggaaactgga gggggacctg aaactggccc aagagtccac aatggacata gaaaacgata
     2041 aacagcagct cgatgagaaa ctcaaaaaga aagagtttga aatgagcaat ctgcagagca
     2101 agattgagga cgagcaggcc ctcggcatgc agctgcagaa gaagatcaag gagttgcagg
     2161 cccgcatcga ggagctggag gaggaaatcg aggcagagcg ggcctccagg gctaaagcag
     2221 agaagcagcg ctctgacctc tcccgggaac tggaggagat cagcgagagg ctggaagaag
     2281 ctggcggagc cacttcagcc cagatcgaga tgaacaagaa gagagaggct gagttccaga
     2341 aaatgcgcag ggacctggag gaggccacgc tgcagcatga agccacagca gccgctcttc
     2401 ggaagaagca cgcggacagc gtggctgagc tcggggagca gatcgacaac ctgcagcggg
     2461 tgaagcagaa gctggagaag gagaagagcg agctgaagat ggagatcgat gatctcgcta
     2521 gtaacatgga gacagtctcc aaagccaagg ggaaccttga gaagatgtgc cgcaccctgg
     2581 aggaccaact gagtgaagtg aagaccaagg aggaggagca gcagcgcctg atcaatgaac
     2641 tgtcaaccca gaaggcacgc ttgcacacag agtcaggcga gttaatcgaa ttcccgcggc
     2701 cgccatggcg gccgggagca tgcgacgtcg ggcccaattc gccctatagt gagtcgtatt
     2761 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     2821 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     2881 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     2941 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3001 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3061 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3121 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3181 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3241 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3301 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3361 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3421 tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3481 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3541 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3601 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3661 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3721 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3781 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3841 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3901 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3961 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcaca
//
