LOCUS       sequence-304859-        6175 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Entry vector encoding mKate2-2xFLAG-OGT (O-GlcNAc Transferase; K842M
            variant, catalytic dead).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6175)
  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 6175)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 K842M
FEATURES             Location/Qualifiers
     source          1..6175
                     /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             487..1182
                     /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             1189..1212
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1216..1239
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1255..1279
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4404..4503)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4615..4634)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4626..5435
                     /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(4693..4712)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5525..6113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6014..6033
                     /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 gcaggtttaa aggaaccaat tcagtcgact
      481 ggatccatgg tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
      541 accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
      601 acccagacca tgagaatcaa ggcggtcgag ggcggccctc tccccttcgc cttcgacatc
      661 ctggctacca gcttcatgta cggcagcaaa accttcatca accacaccca gggcatcccc
      721 gacttcttta agcagtcctt ccccgagggc ttcacatggg agagagtcac cacatacgaa
      781 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
      841 aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca
      901 ctcggctggg aggcctccac cgagaccctg taccccgctg acggcggcct ggaaggcaga
      961 gccgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt gaagaccaca
     1021 tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggacaga
     1081 agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct
     1141 gtggccagat actgcgacct ccctagcaaa ctggggcaca gaggcggaga ctacaaagat
     1201 gatgacgaca aaggcgacta caaagacgat gacgacaagg gcatcgatag atcaacaagt
     1261 ttgtacaaaa aagcaggctt tgcgtcttcc gtgggcaacg tggccgacag cacagggtta
     1321 gctgagttgg cacatcgaga atatcaggca ggagattttg aggcagctga gagacactgc
     1381 atgcagctct ggagacaaga gccagacaat actggtgtgc ttttattact ttcatctata
     1441 cacttccagt gtcgaaggct ggacagatct gctcacttta gcactctggc aattaaacag
     1501 aacccccttc tggcagaagc ttattcgaat ttggggaatg tgtacaagga aagagggcag
     1561 ttgcaggagg caattgagca ttatcgacat gcattgcgtc tcaaacctga tttcatcgat
     1621 ggttatatta acctggcagc cgccttggta gcagcgggtg acatggaagg ggcagtacaa
     1681 gcttacgtct ctgctcttca gtacaatcct gatttgtact gtgttcgcag tgacctgggg
     1741 aacctgctca aagccctggg tcgcttggaa gaagccaagg catgttattt gaaagcaatt
     1801 gagacgcaac cgaactttgc agtagcttgg agtaatcttg gctgtgtttt caatgcacaa
     1861 ggggaaattt ggcttgcaat tcatcacttt gaaaaggctg tcacccttga cccaaacttt
     1921 ctggatgctt atatcaattt aggaaatgtc ttgaaagagg cacgcatttt tgacagagct
     1981 gtggcagctt atcttcgtgc cctaagtttg agtccaaatc acgcagtggt gcacggcaac
     2041 ctggcttgtg tatactatga gcaaggcctg atagatctgg caatagacac ctacaggcgg
     2101 gctatcgaac tacaaccaca tttccctgat gcttactgca acctagccaa tgctctcaaa
     2161 gagaagggca gtgttgctga agcagaagat tgttataata cagctctccg tctgtgtccc
     2221 acccatgcag actctctgaa taacctagcc aatatcaaac gagaacaggg aaacattgaa
     2281 gaggcagttc gcttgtatcg taaagcatta gaagtcttcc cagagtttgc tgctgcccat
     2341 tcaaatttag caagtgtact gcagcagcag ggaaaactgc aggaagctct gatgcattat
     2401 aaggaggcta ttcgaatcag tcctaccttt gctgatgcct actctaatat gggaaacact
     2461 ctaaaggaga tgcaggatgt tcagggagcc ttgcagtgtt atacgcgtgc catccaaatt
     2521 aatcctgcat ttgcagatgc acatagcaat ctggcttcca ttcataagga ttcagggaat
     2581 attccagaag ccatagcttc ttaccgcacg gctctgaaac ttaagcctga ttttcctgat
     2641 gcttattgta acttggctca ttgcctgcag attgtctgtg attggacaga ctatgatgag
     2701 cgaatgaaga agttggtcag tattgtggct gaccagttag agaagaatag gttgccttct
     2761 gtgcatcctc atcatagtat gctatatcct ctttctcatg gcttcaggaa ggctattgct
     2821 gagaggcacg gcaacctgtg cttagataag attaatgttc ttcataaacc accatatgaa
     2881 catccaaaag acttgaagct cagtgatggt cggctgcgtg taggatatgt gagttccgac
     2941 tttgggaatc atcctacttc tcaccttatg cagtctattc caggcatgca caatcctgat
     3001 aaatttgagg tgttctgtta tgccctgagc ccagacgatg gcacaaactt ccgagtgaag
     3061 gtgatggcag aagccaatca tttcattgat ctttctcaga ttccatgcaa tggaaaagca
     3121 gctgatcgca tccatcagga tggaattcat atccttgtaa atatgaatgg ctatactaag
     3181 ggcgctcgaa atgagctttt tgctctcagg ccagctccta ttcaggcaat gtggctggga
     3241 taccctggga cgagtggtgc gcttttcatg gattatatta tcactgatca ggaaacttcg
     3301 ccagctgaag ttgctgagca gtattccgag aaattggctt atatgcccca cacttttttt
     3361 attggtgatc atgctaatat gttccctcac ctgaagaaaa aagcagtcat cgattttaag
     3421 tccaatgggc acatttatga caatcggata gttctgaatg gcatcgacct caaagcattt
     3481 cttgatagtc taccagatgt gaaaattgtc aagatgaagt gtcctgatgg aggagacaat
     3541 gcagatagca gtaacacagc tcttaatatg cctgttattc ctatgaatac tattgcagaa
     3601 gcagttattg aaatgattaa ccgaggacag attcaaataa caattaatgg attcagtatt
     3661 agcaatggac tggcaactac tcagatcaac aataaggctg caactggaga ggaggttccc
     3721 cgtaccatta ttgtaaccac ccgttctcag tacgggttac cagaagatgc catcgtatac
     3781 tgtaacttta atcagttgta tatgattgac ccttctactt tgcagatgtg ggcaaacatt
     3841 ctgaagcgtg ttcccaatag tgtactctgg ctgttgcgtt ttccagcagt aggagaacct
     3901 aatattcaac agtatgcaca aaacatgggc ctgccccaga accgtatcat tttttcacct
     3961 gttgctccta aagaggaaca cgtcaggaga ggccagctgg ctgatgtctg cttggacact
     4021 ccactctgta atgggcacac cacagggatg gatgtcctct gggcagggac ccccatggtg
     4081 actatgccag gagagactct tgcttctcga gttgcagcat cccagctcac ttgcttaggt
     4141 tgtcttgagc ttattgctaa aaacagacaa gaatatgaag acatagctgt gaagctggga
     4201 actgatctag aatacctgaa gaaagttcgt ggcaaagtct ggaagcaaag aatatctagc
     4261 cctctgttca acaccaaaca atacacaatg gaactagagc ggctctatct acagatgtgg
     4321 gagcattatg cagctggcaa caaacctgac cacatgatta agcctgttga agtcactgag
     4381 tcagcataac tcgagatatc tagacccagc tttcttgtac aaagttggca ttataagaaa
     4441 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     4501 ttgccatcca gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     4561 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     4621 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     4681 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     4741 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     4801 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     4861 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     4921 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     4981 ttgatgcgct ggcagtgtcc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     5041 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     5101 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     5161 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     5221 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     5281 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     5341 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     5401 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     5461 attattcaga ttgggccccg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5521 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5581 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5641 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5701 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5761 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5821 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5881 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5941 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6001 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6061 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6121 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
