LOCUS       sequence-238588-        3930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making constitutively active CamKII gamma RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3930)
  AUTHORS   Miao YL, Stein P, Jefferson WN, Padilla-Banks E, Williams CJ
  TITLE     Calcium influx-mediated signaling is required for complete mouse egg
            activation.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Mar 13;109(11):4169-74. Epub 2012 Feb
            27.
  PUBMED    22371584
REFERENCE   2  (bases 1 to 3930)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2012-03-13"; volume: "109(11)"; pages: 
            "4169-74. Epub 2012 Feb 27"
COMMENT     Sequence Label: pIVT/CamKII gamma CA
FEATURES             Location/Qualifiers
     source          1..3930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    80..101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(116..121,122..139,140..146)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    154..170
                     /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     159..181
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     178..194
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     178..194
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        218..236
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     218..235
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     248..267
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        248..266
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           309..351
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1325..1450
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1508..1525)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1508..1524)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1517..1539)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1733..1752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1852..1874
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1912..1930)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1998..2102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2103..2171,2172..2963)
                     /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(2321..2340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3134..3722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3623..3642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3876..3893
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
       61 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      121 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      181 gaaacagcta tgaccatgat tacgccaagc tccaacgatt taggtgacac tatagaagag
      241 aaggaattaa tacgactcac tatagggaga gagagagaat taccctcact aaagggagga
      301 gaagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg gcagatcaag
      361 cttgcatgcc tgcaggccag tatggccacc accgccacct gcacccgctt caccgacgac
      421 taccagcttt tcgaggagct cggcaagggt gccttctcag tggtccggag gtgtgtaaag
      481 aaaacatcta cgcaggaata tgctgcaaaa atcatcaata ccaagaagtt gtccgcccga
      541 gatcatcaga aactagaacg agaggcccgg atatgccgac ttctgaaaca tccaaacatt
      601 gtgcgtctcc atgacagtat ttctgaagaa gggtttcact acctcgtgtt tgaccttgtt
      661 accggagggg agttgtttga agacattgtg gccagagagt actacagtga agctgatgcc
      721 agccactgta tacatcagat cctggagagt gttaaccaca tccaccagca tgacatcgtc
      781 caccgggatc tgaagcctga gaacttgctg ctggcgagta aatgcaaggg tgctgcggtc
      841 aagctagctg attttggcct ggccatcgaa gtacagggag agcagcaggc ttggtttggt
      901 tttgccggca ccccaggtta cttgtcccct gaggtcttga ggaaagatcc ctatggaaaa
      961 cctgtggaca tctgggcctg cggggtcatc ctgtatatcc tcctggtggg ctaccctccc
     1021 ttctgggatg aggatcagca caagctgtat cagcagatca aagctggagc ctacgatttc
     1081 ccatcaccag aatgggacac agtcactcct gaagctaaga acttgatcaa ccagatgctg
     1141 accataaacc ctgcaaagcg catcacggcc gaccaggctc tcaagcaccc atgggtctgt
     1201 caacggtcta cggtggcatc catgatgcat cgccaagaga cggtagagtg cttatagtaa
     1261 ggatccccgg gtaccgagct cgaattcgcc cttctagtga ctgactagga tctggttacc
     1321 actaaaccag cctcaagaac acccgaatgg agtctctaag ctacataata ccaacttaca
     1381 ctttacaaaa tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa taaaaagaaa
     1441 gtttcttcac attctaaaaa aaaaaaaaaa aaaaaaaaaa aaaccccccc cccccaaggg
     1501 cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1561 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1621 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     1681 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     1741 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     1801 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1861 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     1921 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1981 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2041 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2101 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2161 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2221 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2281 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2341 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2401 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2461 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2521 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2581 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2641 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2701 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2761 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2821 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2881 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2941 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3001 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3061 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3241 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3781 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3841 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3901 aagagcgccc aatacgcaaa ccgcctctcc
//
