LOCUS       sequence-239433-        2115 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  SB-transposon plasmid for stable expression of unphosphorylatable 
            ""dead"" version of AktAR2 variant of FRET-based AKT activity 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2115)
  TITLE     Eric Davis SB transposon plasmids
REFERENCE   2  (bases 1 to 2115)
  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"
COMMENT     Sequence Label: pSBbi-Pur-AktAR2-D
FEATURES             Location/Qualifiers
     source          1..2115
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             22..39
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             43..75
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(79..87,88..102)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(109..111,112..114,115..825)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1378..2112
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
ORIGIN
        1 gccaccatgg tgcggggttc tcatcatcat catcatcatg gtatggctag catgactggt
       61 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatcccat ggtgagcaag
      121 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      181 ggccacaggt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      241 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      301 ctgagctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      361 ttcaagtccg ccatgcccga aggctacgtc caggagcgta ccatcttctt caaggacgac
      421 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      481 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      541 aacgccatcc acggcaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
      601 aacttcggcc tcaactgcaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      661 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
      721 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      781 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcgcat gcataagttt
      841 tctcaagaac agatcggcga aaacattgtg tgcagggtca tttgtaccac gggtcaaatt
      901 cccatccgag atttgtcagc tgatatttca caagtgctta aggaaaaacg atccataaag
      961 aaagtttgga catttggtag aaacccagcc tgtgactatc atttaggaaa catttcaaga
     1021 ctgtcaaata agcatttcca aatactacta ggagaagacg gtaacctttt attgaatgac
     1081 atttccacta atgggacctg gttaaatggg caaaaagtcg agaagaacag caatcagtta
     1141 ctgtctcaag gtgatgaaat aaccgttggt gtaggcgtgg aatcagatat tttatctctg
     1201 gtcattttca taaacgacaa atttaagcag tgcctcgagc agaacaaagt tgatcgctct
     1261 gcaggtaagc caggcagcgg cgagggcagc accaagggcc tggtcgaccc tcgtccgcgc
     1321 tcgtgcgtgt ggccggaccc caggccggag tttggaggta ccggcggcag cgagctcatg
     1381 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1441 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     1501 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1561 gacgagctgt acaagggagg taccggtgga tctatggtga gcaagggcga ggagctgttc
     1621 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1681 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     1741 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctt
     1801 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1861 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1921 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1981 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2041 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     2101 cacaacatcg agtaa
//
