LOCUS       sequence-304861-        6176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector encoding mKate2-2xFLAG-OGT (O-GlcNAc Transferase; 5N5A 
            variant, no HCF-1 cleavage).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6176)
  AUTHORS   Levine ZG, Potter SC, Joiner CM, Fei GQ, Nabet B, Sonnett M, Zachara
            NE, Gray NS, Paulo JA, Walker S
  TITLE     Mammalian cell proliferation requires noncatalytic functions of 
            O-GlcNAc transferase.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jan 26;118(4). pii: 2016778118. doi: 
            10.1073/pnas.2016778118.
  PUBMED    33419956
REFERENCE   2  (bases 1 to 6176)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-01-26"; volume: "118(4). pii: 
            2016778118. doi"; pages: " 10.1073/pnas.2016778118"
COMMENT     Sequence Label: pENTR1A mKate2-hu_ncOGT 5N5A
FEATURES             Location/Qualifiers
     source          1..6176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             488..1183
                     /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             1190..1213
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1217..1240
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1256..1280
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4405..4504)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4616..4635)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4627..5436
                     /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
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4694..4713)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5526..6114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6015..6034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccatg gtgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg
      541 caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
      601 cacccagacc atgagaatca aggcggtcga gggcggccct ctccccttcg ccttcgacat
      661 cctggctacc agcttcatgt acggcagcaa aaccttcatc aaccacaccc agggcatccc
      721 cgacttcttt aagcagtcct tccccgaggg cttcacatgg gagagagtca ccacatacga
      781 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
      841 caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac
      901 actcggctgg gaggcctcca ccgagaccct gtaccccgct gacggcggcc tggaaggcag
      961 agccgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact tgaagaccac
     1021 atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggacag
     1081 aagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc acgaggtggc
     1141 tgtggccaga tactgcgacc tccctagcaa actggggcac agaggcggag actacaaaga
     1201 tgatgacgac aaaggcgact acaaagacga tgacgacaag ggcatcgata gatcaacaag
     1261 tttgtacaaa aaagcaggct ttgcgtcttc cgtgggcaac gtggccgaca gcacagggtt
     1321 agctgagttg gcacatcgag aatatcaggc aggagatttt gaggcagctg agagacactg
     1381 catgcagctc tggagacaag agccagacaa tactggtgtg cttttattac tttcatctat
     1441 acacttccag tgtcgaaggc tggacagatc tgctcacttt agcactctgg caattaaaca
     1501 gaaccccctt ctggcagaag cttattcgaa tttggggaat gtgtacaagg aaagagggca
     1561 gttgcaggag gcaattgagc attatcgaca tgcattgcgt ctcaaacctg atttcatcga
     1621 tggttatatt aacctggcag ccgccttggt agcagcgggt gacatggaag gggcagtaca
     1681 agcttacgtc tctgctcttc agtacaatcc tgatttgtac tgtgttcgca gtgacctggg
     1741 gaacctgctc aaagccctgg gtcgcttgga agaagccaag gcatgttatt tgaaagcaat
     1801 tgagacgcaa ccgaactttg cagtagcttg gagtaatctt ggctgtgttt tcaatgcaca
     1861 aggggaaatt tggcttgcaa ttcatcactt tgaaaaggct gtcacccttg acccaaactt
     1921 tctggatgct tatatcaatt taggaaatgt cttgaaagag gcacgcattt ttgacagagc
     1981 tgtggcagct tatcttcgtg ccctaagttt gagtccaaat cacgcagtgg tgcacggcaa
     2041 cctggcttgt gtatactatg agcaaggcct gatagatctg gcaatagaca cctacaggcg
     2101 ggctatcgaa ctacaaccac atttccctga tgcttactgc aacctagcca atgctctcaa
     2161 agagaagggc agtgttgctg aagcagaaga ttgttataat acagctctcc gtctgtgtcc
     2221 cacccatgca gactctctga atgcactagc caatatcaaa cgagaacagg gaaacattga
     2281 agaggcagtt cgcttgtatc gtaaagcatt agaagtcttc ccagagtttg ctgctgccca
     2341 ttcagcatta gcaagtgtac tgcagcagca gggaaaactg caggaagctc tgatgcatta
     2401 taaggaggct attcgaatca gtcctacctt tgctgatgcc tactctgcaa tgggaaacac
     2461 tctaaaggag atgcaggatg ttcagggagc cttgcagtgt tatacgcgtg ccatccaaat
     2521 taatcctgca tttgcagatg cacatagcgc actggcttcc attcataagg attcagggaa
     2581 tattccagaa gccatagctt cttaccgcac ggctctgaaa cttaagcctg attttcctga
     2641 tgcttattgt gcattggctc attgcctgca gattgtctgt gattggacag actatgatga
     2701 gcgaatgaag aagttggtca gtattgtggc tgaccagtta gagaagaata ggttgccttc
     2761 tgtgcatcct catcatagta tgctatatcc tctttctcat ggcttcagga aggctattgc
     2821 tgagaggcac ggcaacctgt gcttagataa gattaatgtt cttcataaac caccatatga
     2881 acatccaaaa gacttgaagc tcagtgatgg tcggctgcgt gtaggatatg tgagttccga
     2941 ctttgggaat catcctactt ctcaccttat gcagtctatt ccaggcatgc acaatcctga
     3001 taaatttgag gtgttctgtt atgccctgag cccagacgat ggcacaaact tccgagtgaa
     3061 ggtgatggca gaagccaatc atttcattga tctttctcag attccatgca atggaaaagc
     3121 agctgatcgc atccatcagg atggaattca tatccttgta aatatgaatg gctatactaa
     3181 gggcgctcga aatgagcttt ttgctctcag gccagctcct attcaggcaa tgtggctggg
     3241 ataccctggg acgagtggtg cgcttttcat ggattatatt atcactgatc aggaaacttc
     3301 gccagctgaa gttgctgagc agtattccga gaaattggct tatatgcccc acactttttt
     3361 tattggtgat catgctaata tgttccctca cctgaagaaa aaagcagtca tcgattttaa
     3421 gtccaatggg cacatttatg acaatcggat agttctgaat ggcatcgacc tcaaagcatt
     3481 tcttgatagt ctaccagatg tgaaaattgt caagatgaag tgtcctgatg gaggagacaa
     3541 tgcagatagc agtaacacag ctcttaatat gcctgttatt cctatgaata ctattgcaga
     3601 agcagttatt gaaatgatta accgaggaca gattcaaata acaattaatg gattcagtat
     3661 tagcaatgga ctggcaacta ctcagatcaa caataaggct gcaactggag aggaggttcc
     3721 ccgtaccatt attgtaacca cccgttctca gtacgggtta ccagaagatg ccatcgtata
     3781 ctgtaacttt aatcagttgt ataaaattga cccttctact ttgcagatgt gggcaaacat
     3841 tctgaagcgt gttcccaata gtgtactctg gctgttgcgt tttccagcag taggagaacc
     3901 taatattcaa cagtatgcac aaaacatggg cctgccccag aaccgtatca ttttttcacc
     3961 tgttgctcct aaagaggaac acgtcaggag aggccagctg gctgatgtct gcttggacac
     4021 tccactctgt aatgggcaca ccacagggat ggatgtcctc tgggcaggga cccccatggt
     4081 gactatgcca ggagagactc ttgcttctcg agttgcagca tcccagctca cttgcttagg
     4141 ttgtcttgag cttattgcta aaaacagaca agaatatgaa gacatagctg tgaagctggg
     4201 aactgatcta gaatacctga agaaagttcg tggcaaagtc tggaagcaaa gaatatctag
     4261 ccctctgttc aacaccaaac aatacacaat ggaactagag cggctctatc tacagatgtg
     4321 ggagcattat gcagctggca acaaacctga ccacatgatt aagcctgttg aagtcactga
     4381 gtcagcataa ctcgagatat ctagacccag ctttcttgta caaagttggc attataagaa
     4441 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     4501 tttgccatcc agctgcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     4561 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     4621 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     4681 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     4741 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     4801 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     4861 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     4921 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     4981 gttgatgcgc tggcagtgtc cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     5041 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     5101 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     5161 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     5221 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     5281 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     5341 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     5401 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     5461 cattattcag attgggcccc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5641 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
