LOCUS       sequence-380207-        4066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to produce ISH probe for mMyh7 gene with CDS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4066)
  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 4066)
  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-mMyh7CDS
FEATURES             Location/Qualifiers
     source          1..4066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     78..97
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(316..420)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(498..953)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(635..656)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     847..866
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1079..1101
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1093..1110
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1094..1110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1117..1136
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1117..1135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1166..1175
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1170..1179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2324..2342)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2325..2342)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2360..2376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2360..2376)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2373..2395)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2384..2400
                     /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(2408..2414,2415..2432,2433..2438))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2453..2474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2591..2608)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2762..3350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2842..2861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3521..4066,1..246,247..315))
                     /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 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
       61 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
      121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
      181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
      241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
      301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
      361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
      421 cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga
      481 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
      541 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      601 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
      661 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
      721 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
      781 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
      841 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
      901 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
      961 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     1021 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     1081 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     1141 ttgggcccga cgtcgcatgc tcccggccgc catggcggcc gcgggaattc gattgaacca
     1201 gacggcactg aagaggctga caaatcagcc tacctcatgg ggctgaactc agctgacctg
     1261 cttaaggggc tgtgccatcc tagagtcaaa gtgggcaacg agtacgtcac caaggggcag
     1321 aatgttcagc aggtgtcata tgccatcggg gcactggcca agtcagtgta cgagaagatg
     1381 ttcaactgga tggtgacacg catcaacgca accctggaga ccaagcagcc gcgccagtac
     1441 ttcataggtg tcctggacat tgccggcttt gagatcttcg atttcaacag ctttgagcag
     1501 ctgtgcatca acttcaccaa tgagaagctg cagcagttct tcaaccacca catgttcgtg
     1561 ctggagcagg aggagtacaa gaaggagggc attgagtgga cattcataga cttcggcatg
     1621 gacttgcagg cctgcattga cctcatcgag aagcccatgg gcatcatgtc catccttgag
     1681 gaggagtgca tgttccccaa ggccacagac atgaccttca aggccaagct gtacgacaac
     1741 cacctgggca agtccaacaa cttccagaag cctcgcaatg tcaaggggaa gcaggaagcc
     1801 cacttctcct tggtccacta tgctggcact gtggactaca atatcctggg ctggctacag
     1861 aagaacaagg acccactcaa tgagacagtg gtgggtttgt accagaagtc ttccctcaag
     1921 ctgctcagca atctatttgc caactatgct ggagctgatg ccccggcgga caaaggcaaa
     1981 ggcaaggcaa agaaaggctc atcctttcag accgtgtctg ctctgcacag agaaaatctg
     2041 aacaaactta tgacaaactt gcgctccacg caccctcact ttgtacgctg catcatcccc
     2101 agtgagacaa agtcgccagg ggtgatggac agtcccctgg tcatgcacca gctgcgatgc
     2161 aatggcgtgc tggagggtat ccgcatctgc aggaagggct ttcccaaccg cattctctat
     2221 ggggacttcc ggcagaggta tcgaatcact agtgaattcg cggccgcctg caggtcgacc
     2281 atatgggaga gctcccaacg cgttggatgc atagcttgag tattctatag tgtcacctaa
     2341 atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2401 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2461 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2521 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2581 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2641 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2701 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2761 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2821 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2881 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2941 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3001 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3061 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3121 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3181 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3241 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3301 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3361 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3421 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3481 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3541 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3601 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3661 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3721 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3781 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3841 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3901 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3961 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4021 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgac
//
