LOCUS       sequence-380215-        4027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh13 gene with CDS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4027)
  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 4027)
  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-mMyh13CDS
FEATURES             Location/Qualifiers
     source          1..4027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     289..308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(527..631)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(709..1164)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(846..867)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1058..1077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1290..1312
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1304..1321
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1305..1321
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1328..1347
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1328..1346
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1377..1386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1381..1390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2496..2514)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2497..2514)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2532..2548)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2532..2548)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2545..2567)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2556..2572
                     /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(2580..2586,2587..2604,2605..2610))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2625..2646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2763..2780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2934..3522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3014..3033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3693..4027,1..457,458..526))
                     /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 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
       61 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
      181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
      241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
      301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
      361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
      421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
      481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
      541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
      601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgatg
      661 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
      721 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
      781 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      841 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
      901 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      961 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1021 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1081 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1141 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
     1201 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     1261 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     1321 tgaattgtaa tacgactcac tatagggcga attgggcccg acgtcgcatg ctcccggccg
     1381 ccatggcggc cgcgggaatt cgattagtgc atgttcccta aggccacaga cacctccttc
     1441 aagaacaagt tgtatgacca gcacctggga aagtccaaca acttccagaa gcctaaaccc
     1501 accaaaggca aggccgaagc ccacttctcc ctggtgcact atgcgggcac tgtggattac
     1561 aacatcgctg gctggctgga taagaacaag gacccactga atgagaccgt ggtggggctg
     1621 taccagaagt cctccctgaa gttgttgtcc ttcctctttt ccaactacgc aggagcagaa
     1681 gcaggtgact ccgcaggagg caagaaggga ggaaagaaga agggctcctc cttccagact
     1741 gtgtctgctg tgttcaggga aaacttaaac aaattgatga ccaacctaag gagcacccat
     1801 cctcactttg tgcgatgcct gattcccaat gagaccaaga ctcctggtgt gatggaccac
     1861 tgcttggtca tgcatcagct gcgctgtaat ggggtcctgg agggcatccg gatctgcagg
     1921 aaagggttcc ccagtcgaat cctctatgct gacttcaaac aacggtaccg gatccttaac
     1981 gccagtgcca tcccggaagg gcagttcatc gacagcaaga acgcctcgga gaagctcctg
     2041 aactccatcg atgtggaccg agagcagttc cgattcggcc ataccaaggt gtttttcaaa
     2101 gctgggctcc tggggcttct ggaagagatg agagacgaga agctggtgac cctaatgact
     2161 cgcacacagg cagtgtgcag gggctacctg atgagggtgg agttcaagaa aatgatggag
     2221 aggagggagt ccatcttctg catccagtac aatgttcgtt ccttcatgaa cgtcaagcac
     2281 tggccctgga tgaacctgtt cttcaagatc aagcccctgc tgaagagcgc ggaggcggag
     2341 aaggagatgg ccaccatgaa ggaggacttc gaacgagcca aggaagacct ggctagatct
     2401 gaggctcgtc ggaagaatca ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga
     2461 gagctcccaa cgcgttggat gcatagcttg agtattctat agtgtcacct aaatagcttg
     2521 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2581 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2641 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2701 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4021 gtgtcac
//
