LOCUS       sequence-380188-        3641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh2 gene with CDS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3641)
  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 3641)
  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-mMyh2CDS
FEATURES             Location/Qualifiers
     source          1..3641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      492..1080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     981..1000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1234..1251
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1368..1389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1404..1409,1410..1427,1428..1434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1442..1458
                     /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     1447..1469
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1466..1482
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1466..1482
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1500..1518
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1500..1517
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      2238..2247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2242..2251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2281..2300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2282..2300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2307..2324)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2307..2323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2316..2338)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2464..2919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2551..2570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2761..2782
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2997..3101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3102..3170,3171..3641,1..321)
                     /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(3320..3339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
       61 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      121 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      181 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      241 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      301 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      361 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      421 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
      481 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      541 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      601 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      661 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      721 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      781 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      841 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      901 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      961 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1021 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1081 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1141 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     1201 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     1261 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     1321 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     1381 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcta
     1501 tttaggtgac actatagaat actcaagcta tgcatccaac gcgttgggag ctctcccata
     1561 tggtcgacct gcaggcggcc gcgaattcac tagtgattgg agctgaagaa ggagcaggac
     1621 actagcgccc acctggagcg gatgaagaag aacatggagc agaccgtgaa ggacctgcag
     1681 ctgcgtctgg atgaggctga gcagctggcg ctgaagggcg gcaagaagca gatccagaaa
     1741 ctggaggcca gggtgcgtga actggagggt gaggtagaga gtgagcagaa gcggaatgct
     1801 gaggccgtca aaggtctgcg caaacacgag agacgaatga aggagcttac ttaccagaca
     1861 gaagaagacc gaaaaaatat tctcaggctt caggatttgg tggataaact ccaggcaaaa
     1921 gtgaaatctt acaagagaca agctgaggag gctgaggaac aatccaacac aaatctatcc
     1981 aagttccgca agatccagca cgagctggag gaagccgagg agcgggctga catcgcggag
     2041 tcccaggtca acaagctgcg ggtgaagagc cgggaggttc acaccaaaat cataagcgaa
     2101 gagtaaggct gtcccgatgc tgtggaatga ccgaagcgag gcacaaaatg tgaaaccttt
     2161 ggtcatgcgc ctgtgtgatt ctattccatc ctgttgtaag gaaataaaaa gcccaagttc
     2221 ctaatcgaat tcccgcggcc gccatggcgg ccgggagcat gcgacgtcgg gcccaattcg
     2281 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2341 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2401 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2461 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2521 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2581 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2641 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2701 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2761 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2821 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2881 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     2941 tacgcatctg tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg
     3001 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3061 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3121 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3181 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3241 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3301 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3361 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3421 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3481 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3541 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3601 tgaatgaagc cataccaaac gacgagcgtg acaccacgat g
//
