LOCUS       sequence-152953-        3370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hemagglutinin (HA) epitope fused to C-terminus of mKate2 without pA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3370)
  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 3370)
  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: p3E-mKate2-HA no-pA
FEATURES             Location/Qualifiers
     source          1..3370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             17..712
                     /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"
     CDS             713..739
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(746..841)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(878..897)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(879..897)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(902..918)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(902..918)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1020..1039)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1031..1840
                     /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(1098..1117)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1987..2575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2476..2495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2729..2746
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2905..2932)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3024..3110)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3159..3181
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3173..3190
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3174..3190
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(3261..3370,1..14))
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cttgtacaaa gtggccatgg tgagcgagct gattaaggag aacatgcaca tgaagctgta
       61 catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc
      121 ctacgagggc acccagacca tgagaatcaa ggcggtcgag ggcggccctc tccccttcgc
      181 cttcgacatc ctggctacca gcttcatgta cggcagcaaa accttcatca accacaccca
      241 gggcatcccc gacttcttta agcagtcctt ccccgagggc ttcacatggg agagagtcac
      301 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
      361 cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca
      421 gaagaaaaca ctcggctggg aggcctccac cgagaccctg taccccgctg acggcggcct
      481 ggaaggcaga gccgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt
      541 gaagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta
      601 tgtggacaga agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca
      661 cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca gatatccgta
      721 cgacgtacca gactacgcag catagcaact ttattataca aagttggcat tataaaaaag
      781 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
      841 tggagctcca tggtagcgtt aacgcggccg cgatatcccc tatagtgagt cgtattacat
      901 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
      961 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1021 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1081 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1141 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1201 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1261 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1321 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1381 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     1441 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     1501 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     1561 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     1621 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     1681 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     1741 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     1801 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     1861 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     1921 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1981 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2101 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2341 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     2401 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2641 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2701 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2761 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2821 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
     2881 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
     2941 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
     3001 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     3061 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
     3121 gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac
     3181 gacggccagt cttaagctcg ggccctgcag ctctagagct cgaattctac aggtcactaa
     3241 taccatctaa gtagttgatt catagtgact gcatatgttg tgttttacag tattatgtag
     3301 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     3361 gttcaacttt
//
