LOCUS       sequence-153013-        3246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  red fluorophore without stop codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3246)
  AUTHORS   Fowler DK, Stewart S, Seredick S, Eisen JS, Stankunas K, Washbourne 
            P
  TITLE     A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate 
            Expression Constructs with Diverse Research Applications.
  JOURNAL   PLoS One. 2016 Aug 8;11(8):e0159277. doi: 
            10.1371/journal.pone.0159277. eCollection 2016.
  PUBMED    27500400
REFERENCE   2  (bases 1 to 3246)
  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: "PLoS One.";
            date: "2016-08-8"; volume: "11(8):e0159277. doi"; pages: " 
            10.1371/journal.pone.0159277. eCollection 2016"
COMMENT     Sequence Label: pME-mKate2 no stop
FEATURES             Location/Qualifiers
     source          1..3246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..718
                     /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"
     protein_bind    complement(720..819)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(836..855)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(837..855)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(860..876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(860..876)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(978..997)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             989..1798
                     /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(1056..1075)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1945..2533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2434..2453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2687..2704
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2863..2890)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2982..3068)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3117..3139
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3131..3148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3132..3148
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(3164..3246,1..17)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gtacaaaaaa gcaggctcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa
       61 gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg
      121 caagccctac gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc
      181 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca
      241 cacccagggc atccccgact tctttaagca gtccttcccc gagggcttca catgggagag
      301 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
      361 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt
      421 gatgcagaag aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg
      481 cggcctggaa ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg
      541 caacttgaag accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt
      601 ctactatgtg gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga
      661 gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagata
      721 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
      781 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
      841 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
      901 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
      961 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     1021 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1081 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     1141 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1201 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1261 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     1321 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     1381 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     1441 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     1501 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     1561 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1621 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1681 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1741 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1801 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     1861 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     1921 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2101 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2161 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2221 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2281 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2341 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2401 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2461 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2521 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2581 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2641 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2701 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2761 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2821 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     2881 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     2941 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     3001 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     3061 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     3121 gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt
     3181 ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc
     3241 aacttt
//
