LOCUS       sequence-70388-e        3438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contributes the coding sequence for the mKate2 fluorescent protein 
            as the 5'-module during MultiSite Gateway-cloning of chimeric cDNAs 
            encoding three-part fusion proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3438)
  AUTHORS   Buj R, Iglesias N, Planas AM, Santalucia T
  TITLE     A plasmid toolkit for cloning chimeric cDNAs encoding customized 
            fusion proteins into any Gateway destination expression vector.
  JOURNAL   BMC Mol Biol. 2013 Aug 20;14(1):18. doi: 10.1186/1471-2199-14-18.
  PUBMED    23957834
REFERENCE   2  (bases 1 to 3438)
  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: "BMC Mol 
            Biol."; date: "2013-08-20"; volume: "14(1):18. doi"; pages: " 
            10.1186/1471-2199-14-18"
COMMENT     Sequence Label: pDONR P4-P1R-mKate2
FEATURES             Location/Qualifiers
     source          1..3438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      691..700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             697..1392
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    1405..1470
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    1483..1607
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1679..1698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1680..1698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1703..1719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1703..1719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1821..1840)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1832..2641
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1899..1918)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2788..3376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3277..3296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgcc gccaccatgg tgagcgagct gattaaggag
      721 aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca
      781 tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggcggtcgag
      841 ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaaa
      901 accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt ccccgagggc
      961 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac
     1021 accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca
     1081 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggcctccac cgagaccctg
     1141 taccccgctg acggcggcct ggaaggcaga gccgacatgg ccctgaagct cgtgggcggg
     1201 ggccacctga tctgcaactt gaagaccaca tacagatcca agaaacccgc taagaacctc
     1261 aagatgcccg gcgtctacta tgtggacaga agactggaaa gaatcaagga ggccgacaaa
     1321 gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa
     1381 ctggggcaca gaggtggagg aggttccgga ctcagatctc gagctcaagc ttcgaattct
     1441 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gaacaagttt gtacaaaaaa
     1501 gttgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     1561 aaacagacta cataatactg taaaacacaa catatgcagt cactatgaac caactactta
     1621 gatggtatta gtgacctgta gaattcgagc tctagagctg cagggcggcc gcgatatccc
     1681 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1741 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1801 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1861 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1921 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     1981 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2041 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2101 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2161 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2221 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2281 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2341 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2401 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2461 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2521 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2581 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2641 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2701 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2761 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2821 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2881 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2941 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3001 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3061 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca
     3181 ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3421 gccttttgct cacatgtt
//
