LOCUS       sequence-380205-        3162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh6 gene with UTR sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3162)
  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 3162)
  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-mMyh6UTR
FEATURES             Location/Qualifiers
     source          1..3162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     97..116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     350..367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    484..505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(520..525,526..543,544..550)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    558..574
                     /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     563..585
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     582..598
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     582..598
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             594..1124
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDSGALR
                     AVRQRFLQPRISGLVSQPLLLLFLPVPLSVQLRHCGASFQLWSTIEFPRPPWRPGACDV
                     GPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLR
                     SLNGEWTRPVAAH"
     promoter        616..634
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     616..633
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      875..884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      879..888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(918..937)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(919..937)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(944..961)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(944..960)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(953..975)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1101..1556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1188..1207)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1398..1419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1634..1738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1739..1807,1808..2599)
                     /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(1957..1976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2770..3162,1..196)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
       61 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      301 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      361 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      421 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      481 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      541 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
      601 tgattacgcc aagctattta ggtgacacta tagaatactc aagctatgca tccaacgcgt
      661 tgggagctct cccatatggt cgacctgcag gcggccgcga attcactagt gattctggag
      721 cactgagagc tgtcagacag agatttctcc aacccaggat ctctggattg gtctcccagc
      781 ctctgctact cctcttcctg cctgttcctc tctccgtcca gctgcgccac tgtggtgcct
      841 cgttccagct gtggtccaca atcgaattcc cgcggccgcc atggcggccg ggagcatgcg
      901 acgtcgggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
      961 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1021 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1081 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1141 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1201 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1261 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1321 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1381 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1441 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1501 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     1561 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt
     1621 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1741 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1801 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1861 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1921 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1981 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2041 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2101 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2161 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2221 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2281 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2341 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2401 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2461 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2521 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2581 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2641 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2701 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2761 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2821 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2881 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ct
//
