LOCUS       sequence-238591-        3057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making cRNA appropriate for microinjection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3057)
  AUTHORS   Igarashi H, Knott JG, Schultz RM, Williams CJ
  TITLE     Alterations of PLCbeta1 in mouse eggs change calcium oscillatory 
            behavior following fertilization.
  JOURNAL   Dev Biol. 2007 Dec 1;312(1):321-30. Epub 2007 Sep 26.
  PUBMED    17961538
REFERENCE   2  (bases 1 to 3057)
  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 Biol.";
            date: "2007-12-1"; volume: "312(1)"; pages: "321-30. Epub 2007 Sep 
            26"
COMMENT     Sequence Label: pIVT
FEATURES             Location/Qualifiers
     source          1..3057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..33
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    150..171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(186..191,192..209,210..216)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    224..240
                     /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     229..251
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     248..264
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     248..264
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        288..306
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     288..305
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     318..337
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        318..336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           379..421
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     misc_feature    428..484
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     3'UTR           522..647
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(705..722)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(705..721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(714..736)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(930..949)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1049..1071
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1109..1127)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1195..1299
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1300..1368,1369..2160)
                     /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(1518..1537)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2331..2919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2820..2839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
       61 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      121 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      181 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      241 tttcacacag gaaacagcta tgaccatgat tacgccaagc tccaacgatt taggtgacac
      301 tatagaagag aaggaattaa tacgactcac tatagggaga gagagagaat taccctcact
      361 aaagggagga gaagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg
      421 gcagatcaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
      481 attcgccctt ctagtgactg actaggatct ggttaccact aaaccagcct caagaacacc
      541 cgaatggagt ctctaagcta cataatacca acttacactt tacaaaatgt tgtcccccaa
      601 aatgtagcca ttcgtatctg ctcctaataa aaagaaagtt tcttcacatt ctaaaaaaaa
      661 aaaaaaaaaa aaaaaaaaaa mccccccccc ccaagggcga attcactggc cgtcgtttta
      721 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      781 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      841 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      901 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      961 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     1021 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     1081 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     1141 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1201 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1261 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1321 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1381 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1441 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1501 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1561 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1621 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1681 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1741 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1801 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1861 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1921 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1981 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2041 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2101 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2161 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2221 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2281 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2341 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2401 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2461 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2521 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2581 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2641 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2701 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2761 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2821 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2881 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2941 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3001 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagc
//
