LOCUS       Exported                1688 bp ds-DNA     linear   UNA 02-MAR-2016
DEFINITION  To construct pSP64TSN-mem-mCherry-Plk1 from Original plasmid pTK24: 
            Mem-mCherry-Plk1, Apa1 and Spe1 sites were used.
            
            n fusion
            cloning site for pSP64TSN vector- Apa1-Spe1: 
            atgccatgggggccctcgagactagtgactgacta 
            
             
            
            Primers:
            
            Mem-SP64-fusionF- atgccatgggggccc-atgctgtgctgtatgagaa
            
            Plk-SP64-fusionR -ccgtctcaaggcctcctaa?actagtgactgacta
            
             (primer: tagtcagtcactagtttaggaggccttgagacgg )  
            
            
            For Membrane-mCherry-LOV, PLK1 was replaced to LOV by Not1 and Spe1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1688)
  AUTHORS   Mamiko
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Nov 1, 2019 from SnapGene Viewer 5.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..1688
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     misc_feature    1..64
                     /label=pSP64-vector sequence
     5'UTR           6..48
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     misc_feature    74..133
                     /label=membrane-targeted domain
     CDS             140..847
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     451..461
                     /label=mCherry-Cf1
     primer_bind     601..620
                     /label=mCherry-Cf1
     primer_bind     637..655
                     /label=mCherry-Cf2
     primer_bind     864..905
                     /label=InFus-LOVf
     misc_feature    887..913
                     /label=9aa-linker
     misc_feature    914..1354
                     /label=LovPep
     primer_bind     complement(1335..1376)
                     /label=InFus-LOVr
     misc_feature    1355
                     /label=Human-PLK1
     misc_feature    1356..1688
                     /label=pSP64-vector sequence
     primer_bind     complement(1629..1645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcttgcttgt tctttttgca gaagctcaga ataaacgctc aactttggca gatctgatca
       61 tgccatgggg cccatgctgt gctgtatgag aagaaccaaa caggttgaaa aaaatgatga
      121 cgaccaaaag attggcggca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
      181 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
      241 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      301 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
      361 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
      421 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
      481 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
      541 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
      601 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
      661 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
      721 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
      781 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
      841 gtacaagtcc ggaggggatc ggtggagctc caccggtggc ggccgcggtg gatctggagg
      901 ttcaggtgga agtcctaggt tggctactac acttgaacgt attgagaaga actttgtcat
      961 tactgaccca agattgccag ataatcccat tatattcgcg tccgatagtt tcttgcagtt
     1021 gacagaatat agccgtgaag aaattttggg aagaaactgc aggtttctac aaggtcctga
     1081 aactgatcgc gcgacagtga gaaaaattag agatgccata gataaccaaa cagaggtcac
     1141 tgttcagctg attaattata caaagagtgg taaaaagttc tggaacctct ttcacttgca
     1201 gcctatgcga gatcagaagg gagatgtcca gtactttatt ggggttcagt tggatggaac
     1261 tgagcatgtc cgagatgctg ccgagagaga ggcagtcatg ctgattaaga aaactgcaga
     1321 agagattgat aaagcggtgg atacctgggt gtaaactagt gactgactag gatctggtta
     1381 ccactaaacc agcctcaaga acacccgaat ggagtctcta agctacataa taccaactta
     1441 cactttacaa aatgttgtcc ccccaaaatg tagccattcg tatctgctcc taataaaaaa
     1501 gaaagtttct tcacattcta aaaaaaaaaa aaaaaaaaaa aaaaaaaaac cccccccccc
     1561 cccccccccc ccccccccct ggaggtcgac tctanaggat ccccgggcga gctcgaattc
     1621 gtaatcatgt catagctgtt tcctgtgtga aattgttatc ccgctcacaa ttccncacaa
     1681 catacgag
//
