LOCUS Exported 7362 bp ds-DNA circular SYN 08-DEC-2022 DEFINITION For mammalian expression of beta-catenin with 4 O-GlcNAc sites mutated (S23A, T40A, T41A and T112A) and GFP-tag on its N-terminus.. ACCESSION . VERSION . KEYWORDS pEGFP-beta-catenin_S23A,T40A,T41A,T112A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7362) TITLE Aptamers Increase O-GlcNAc on a Single Protein to Regulate Wnt Signaling JOURNAL Unpublished REFERENCE 2 (bases 1 to 7362) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 8, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7362 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 72..375 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 376..579 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 530..550 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 618..627 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 624..1340 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 624..1340 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(669..690) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(930..949) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1277..1298 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4161..4282 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4198..4217) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4252..4271 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4289..4744) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4426..4447) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4638..4657 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4771..4875 /gene="bla" /label=AmpR promoter promoter 4877..5234 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4900..4920) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5085..5220 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5147..5166 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5269..6063 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5323..5342) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5933..5952 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6251..6270) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6295..6342 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6671..7259 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7160..7179 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat 61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt 421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac 481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc 541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc 601 cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg 661 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 721 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 781 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca 841 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 901 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 961 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1021 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1081 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 1141 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 1201 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca 1261 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 1321 gcatggacga gctgtacaag tccgggatcc tggcaaccca agctgacttg atggagttgg 1381 atatggccat ggagccagac agaaaagctg cagtcgcaca ttggcagcag cagtcatatc 1441 tggactctgg tatccattcc ggtgccgctg caactgctcc ctctttgagt ggcaaaggaa 1501 atcctgaaga ggaagatgtg gacacaacgc aagtcctgta tgagtgggag caagggttct 1561 ctcagtcctt tacccaggag caagttgctg atattgatgg tcaatatgca atgactagag 1621 ctcagagagt gcgtgcagct atgttccccg aaacactgga tgaaggaatg caaatcccat 1681 ccgcacaatt tgatgctgcc catccaacta atgtgcagcg cctggctgag ccatcccaga 1741 tgctaaaaca tgctgttgtt aatttgataa actatcagga tgatgctgaa cttgcaactc 1801 gtgcaatccc agaactgacc aaactgttga atgatgagga ccaggtggtg gtaaacaagg 1861 ctgcggttat ggttcatcag ctatccaaaa aggaagcatc tcgccatgct attatgaggt 1921 ctcctcaaat ggtatctgca attgtgcgta ccatgcaaaa tacaaacgat gtggaaacag 1981 cccgctgtac tgcaggcaca ctacacaatc tctcacatca ccgtgaaggc ttgttggcaa 2041 tcttcaaatc aggaggcatc cctgcgttgg ttaaaatgct tgggtctccg gtggattctg 2101 tgttgttcta tgccattact actcttcaca atctcctgtt acatcaggag ggagccaaaa 2161 tggctgtccg tctggctgga gggctgcaaa aaatggttgc cctgctcaac aagacaaatg 2221 tgaaattctt ggccatcacg acagactgtc ttcagatttt agcctatggc aatcaagaaa 2281 gtaagctgat tattctggca agcggtggac cccaagctct agtaaacata atgaggacct 2341 acacttatga gaaactattg tggaccacaa gtagggtgct gaaggtgttg tcagtctgct 2401 ccagcaacaa acctgctatt gttgaggctg gtgggatgca agctttagga ctccacctta 2461 cagatccaag ccagcgtctt gtccagaact gtctctggac cctgagaaat ttgtcagatg 2521 cagcaaccaa gcaggaggga atggaaggcc ttctaggaac tcttgttcag cttttaggat 2581 cagatgacat taatgttgtg acttgcgctg ccggaatcct ttctaacctt acttgcaaca 2641 attacaagaa caagatgatg gtctgccagg ttggtggcat cgaggctctt gtgcgcacag 2701 ttcttcgggc tggagacagg gaagacatca cagaacctgc tatttgtgcg ctccgtcacc 2761 tcaccagcag acatcaggaa gctgaaatgg ctcaaaatgc agtacgtctc cattacggac 2821 tcccagtggt ggttaaactg ttgcacccac cttcacactg gcctttgatc aaggctactg 2881 ttgggttgat tcgcaatctc gcgctctgcc ccgcaaacca tgccccactg cgtgaacaag 2941 gtgctatccc acggctagtt cagctgctgg ttagagcaca tcaagatacc cagcgacgta 3001 cttccatggg tggaacgcaa cagcagtttg tggagggtgt gcgcatggag gaaatcgttg 3061 agggctgtac tggagccctg catattcttg cacgtgatgt tcacaatcga attgtaatca 3121 ggggtctaaa taccattcca ctatttgtgc agttgttgta ctcccccatt gagaatatcc 3181 agagagtagc tgcgggtgta ctttgtgaac ttgctcaaga caaggaagca gctgaagcaa 3241 ttgaagctga aggtgcaact gcccctttaa cagaactgct tcattccagg aatgagggtg 3301 ttgcaacata tgcagctgca gtgctgttca gaatgtcgga ggacaaacca caagactaca 3361 agaagcgact ttcagttgaa ctgacaagct ctctgttccg gactgagcca atggcttgga 3421 acgagacagc ggatcttgga cttgacattg gtgcccaggg agaacctctt ggataccgcc 3481 cagatgatcc tagctaccgt tctttccact ctggcggata cggtcaggat gccttgggta 3541 tggaccctat gatggaacat gaaatgggtg gccaccaccc tggtgctgac tacccagttg 3601 atggtctgcc agatcttggc catgcccagg accttatgga tgggctgcct ccaggtgaca 3661 gtaatcagtt ggcctggttc gatactgacc tgtaaatcat cctttagctg tatcatctga 3721 atgaacttgc attgattggc ctgtagagtt gctgagaggg ctcgaggggt gggctagtat 3781 ttcagaaagt gcctgacaca ctaaccaagc tgagtttcct atgggaacaa ttgaagtaaa 3841 ctttttgttc tggtcctttt tggtcgagga gtaataatac aaatggattt tgggagtgat 3901 tcaagaaacg aggaatgcac aagaatgaat tgcaagatgg aatttatcaa accctagcct 3961 tgcttgttaa aaatttatta ttttttttaa atctctgtcg gaattctgca gtcgacggta 4021 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg 4081 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 4141 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 4201 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 4261 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc 4321 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc 4381 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 4441 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc 4501 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa 4561 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg 4621 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt 4681 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc 4741 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 4801 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 4861 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt 4921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 4981 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 5041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 5101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 5161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 5221 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 5281 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 5341 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 5401 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca 5461 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 5521 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 5581 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 5641 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 5701 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 5761 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc 5821 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 5881 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 5941 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 6001 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 6061 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 6121 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 6181 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg 6241 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 6301 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt 6361 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg 6421 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca 6481 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt 6541 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 6601 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 6661 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 6721 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 6781 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 6841 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 6901 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 6961 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 7021 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 7081 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 7141 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 7201 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 7261 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 7321 tctttcctgc gttatcccct gattctgtgg ataaccgtat ta //