LOCUS       sequence-379243-        8554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of wild-type full-length human ncOGT. The 
            ncOGT is not epitope-tagged, but is codon-optimized for human 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8554)
  AUTHORS   Zhu Y, Hart GW
  TITLE     Dual-specificity RNA aptamers enable manipulation of target-specific
            O-GlcNAcylation and unveil functions of O-GlcNAc on beta-catenin.
  JOURNAL   Cell. 2022 Dec 30:S0092-8674(22)01529-X. doi: 
            10.1016/j.cell.2022.12.016.
  PUBMED    36626902
REFERENCE   2  (bases 1 to 8554)
  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: "Cell. 2022 
            Dec 30:S0092-8674(22)01529-X. doi: 10.1016/j.cell.2022.12.016."
COMMENT     Sequence Label: pcDNA3.1-OGT_opt
FEATURES             Location/Qualifiers
     source          1..8554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3911..3928)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3917..4141
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4187..4615
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4274..4293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4484..4505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4624..4644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4629..4958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4809..4944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4871..4890
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5025..5819
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5079..5098)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5689..5708
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5993..6114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6030..6049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6084..6103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6163..6179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6163..6179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6176..6198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6187..6203
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6211..6217,6218..6235,6236..6241))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6256..6277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6394..6411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6565..7153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6645..6664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7324..8115,8116..8184))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7947..7966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8185..8289)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8364..8383)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      721 ttcgttaaca gatctatggc cagctctgtg ggcaatgtgg ccgattctac agagcctacc
      781 aagcggatgc tgagctttca gggactggcc gaactggccc acagggaata tcaggccggc
      841 gattttgagg ccgccgagag acactgtatg cagctgtggc ggcaagagcc cgataatact
      901 ggcgttctgc tgctgctgtc cagcatccac ttccagtgca gacggctgga tagaagcgcc
      961 cactttagca ccctggctat caagcagaac cctctgctgg ccgaggccta cagcaacctg
     1021 ggcaacgtgt acaaagagcg gggccagctg caagaggcca tcgagcatta tagacacgcc
     1081 ctgagactga agcccgactt catcgacggc tacatcaatc tggccgctgc tctggttgcc
     1141 gctggcgata tggaaggtgc tgtgcaggcc tatgtgtctg ccctgcagta caaccccgac
     1201 ctgtattgcg tcagaagcga tctgggcaac ctgctgaaag ccctgggcag actggaagag
     1261 gccaaggcct gttacctgaa ggccattgag acacagccca acttcgccgt ggcctggtct
     1321 aatctgggct gcgtgttcaa tgctcagggc gagatctggc tggccatcca ccactttgag
     1381 aaggccgtga ctctggaccc caacttcctg gatgcctata tcaacctggg gaatgtgctg
     1441 aaagaggccc ggatcttcga tagagccgtg gccgcttatc tgcgggccct gtctctgtct
     1501 cctaatcacg ctgtggtgca cggcaacctg gcctgcgtgt actatgagca gggcctgatc
     1561 gacctggcca tcgacacata cagacgggcc atcgaactgc agcctcactt ccctgacgcc
     1621 tactgcaatc tggctaacgc cctgaaagaa aagggcagcg tggccgaagc cgaggactgt
     1681 tacaatactg ccctgcggct gtgccccaca cacgccgatt ctctgaacaa cctggctaac
     1741 atcaagcgcg agcagggcaa catcgaagaa gccgtgcggc tgtacagaaa ggccctggaa
     1801 gtgttccccg agtttgccgc cgctcattct aacctggcaa gcgtgctgca acagcaggga
     1861 aaactccaag aggccctgat gcactacaaa gaagccatcc ggatcagccc caccttcgcc
     1921 gatgcctact ccaatatggg caataccctc aaagaaatgc aggacgtgca gggcgctctg
     1981 cagtgttaca caagggccat ccagatcaat cctgccttcg ctgacgccca cagtaacctg
     2041 gccagcattc acaaggactc cggcaacatc ccagaggcca ttgccagcta cagaacagcc
     2101 ctgaagctga aacctgactt ccccgatgct tactgtaatc tcgcccactg cctgcagatc
     2161 gtgtgcgact ggaccgacta cgacgagcgg atgaagaaac tggtgtccat cgtggccgac
     2221 cagctggaaa agaacagact gcctagcgtg caccctcacc actccatgct gtaccctctg
     2281 agccacggct tcagaaaggc tatcgccgag aggcacggca atctgtgtct ggacaagatc
     2341 aacgtgctgc acaagcctcc atacgagcac cccaaggatc tgaagctgtc cgacggcaga
     2401 ctgagagtgg gctacgtgtc cagcgacttc ggcaatcacc ctaccagcca cctgatgcag
     2461 agcatccccg gcatgcacaa ccctgacaag ttcgaggtgt tctgctacgc tctgagcccc
     2521 gacgacggca ccaacttcag agtgaaagtg atggctgagg ccaaccactt tatcgacctg
     2581 tctcagatcc cctgcaacgg caaagccgcc gacagaatcc accaggacgg aatccacatc
     2641 ctggtcaaca tgaacggcta caccaagggc gccagaaacg agctgtttgc cctcagacct
     2701 gctccaatcc aggccatgtg gctgggctat cctggaacaa gcggagccct gttcatggac
     2761 tacatcatca ccgaccaaga gacaagccct gccgaggtgg ccgagcagta ttctgagaag
     2821 ctggcctaca tgcctcacac ctttttcatc ggcgaccacg ccaacatgtt ccctcacctg
     2881 aagaaaaaag ccgtgatcga cttcaagagc aacggccaca tctacgacaa ccggatcgtg
     2941 ctgaacggaa tcgacctgaa agccttcctg gacagcctgc ctgatgtgaa gatcgtgaag
     3001 atgaagtgcc ccgatggcgg cgacaatgcc gactcttcta acactgccct gaacatgcct
     3061 gtgatcccca tgaacacaat cgccgaggcc gtgattgaga tgatcaaccg gggccagatc
     3121 cagattacca tcaacggctt ctccatctcc aacggcctgg ccaccacaca gatcaacaac
     3181 aaggccgcca ccggcgagga agtgcccaga acaatcatcg tgaccaccag aagccagtac
     3241 ggcctgccag aggatgccat cgtgtactgc aacttcaacc agctgtacaa gatcgacccc
     3301 agcactctgc agatgtgggc caacatcctg aagcgggtgc ccaattctgt gctgtggctg
     3361 ctgagatttc ctgccgtggg cgagcccaac atccagcagt atgcccagaa tatgggcctg
     3421 cctcagaacc ggatcatctt cagccccgtg gctcccaaag aggaacacgt tcgaagagga
     3481 cagctggccg acgtgtgcct ggatacccct ctgtgtaatg gccacaccac cggcatggat
     3541 gttctgtggg ctggcacccc tatggtcacc atgcctggcg aaacactggc ctctagagtg
     3601 gctgccagcc agctgacatg tctgggctgt ctggaactga tcgccaagaa ccggcaagag
     3661 tacgaggata tcgccgtgaa gctgggcacc gacctggaat atctgaagaa agtgcggggc
     3721 aaagtgtgga agcagcggat cagcagccct ctgttcaaca ccaagcagta taccatggaa
     3781 ctggaacggc tgtacctgca aatgtgggag cactacgccg ctggcaacaa gcccgaccac
     3841 atgatcaagc ccgtggaagt gacagagagc gcctgatagt ctagagggcc cgtttaaacc
     3901 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3961 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4021 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4081 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4141 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4201 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4261 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4321 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4381 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4441 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4501 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4561 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4621 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4681 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4801 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4861 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4921 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttatatat
     4981 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5041 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5101 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5161 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5221 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5281 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5341 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5401 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5461 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5521 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5581 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5641 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5701 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5761 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5821 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5881 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5941 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6001 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6061 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6121 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6181 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6241 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6301 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6361 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6421 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6481 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6541 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6601 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6661 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6721 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6781 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6841 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6901 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6961 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7021 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7081 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7141 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7201 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7261 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7321 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7381 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7441 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7501 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7561 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7621 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7681 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7741 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7801 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7861 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7921 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7981 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8041 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8101 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8161 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8221 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8281 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8341 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8401 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8461 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     8521 ctgcttcgcg atgtacgggc cagatatacg cgtt
//
