LOCUS       sequence-380190-        3143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh2 gene with UTR sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3143)
  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 3143)
  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-mMyh2UTR
FEATURES             Location/Qualifiers
     source          1..3143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(535..540,541..558,559..565)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    573..589
                     /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     578..600
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     597..613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     597..613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             609..1085
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDSVPML
                     WNDRSEAQNVKPLVMRLCDSIPSCCKEIKSPSSFASNQSNSRGRHGGREHATSGPIRPI
                     VSRITIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     promoter        631..649
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     631..648
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      871..880
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      875..884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(914..933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(915..933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(940..957)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(940..956)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(949..971)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1097..1552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1184..1203)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1394..1415
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1630..1734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1735..1803,1804..2595)
                     /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(1953..1972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2766..3143,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      541 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      601 aaacagctat gaccatgatt acgccaagct atttaggtga cactatagaa tactcaagct
      661 atgcatccaa cgcgttggga gctctcccat atggtcgacc tgcaggcggc cgcgaattca
      721 ctagtgattc tgtcccgatg ctgtggaatg accgaagcga ggcacaaaat gtgaaacctt
      781 tggtcatgcg cctgtgtgat tctattccat cctgttgtaa ggaaataaaa agcccaagtt
      841 cctttgcaag caatcaatcg aattcccgcg gccgccatgg cggccgggag catgcgacgt
      901 cgggcccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
      961 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1021 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1081 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1141 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1201 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1261 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1321 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1381 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1441 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1501 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     1561 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catcaggtgg cacttttcgg
     1621 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1681 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1741 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1801 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1861 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1921 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1981 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2041 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2101 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2161 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2221 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2281 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2341 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2401 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2461 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2521 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2581 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2881 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3121 acgacctaca ccgaactgag ata
//
