LOCUS Exported 4953 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Used for in vivo monitoring of ubiquitin localization.. ACCESSION . VERSION . KEYWORDS GFP-Ub SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4953) AUTHORS Dantuma NP, Groothuis TA, Salomons FA, Neefjes J TITLE A dynamic ubiquitin equilibrium couples proteasomal activity to chromatin remodeling. JOURNAL J Cell Biol. 2006 Apr 10. 173(1):19-26. PUBMED 16606690 REFERENCE 2 (bases 1 to 4953) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4953 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 613..1329 /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(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" CDS 1369..1593 /codon_start=1 /product="human ubiquitin" /label=ubiquitin /translation="QIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFA GKQLEDGRTLSDYNIQKESTLHLVLRLRGG" polyA_signal 1741..1862 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1778..1797) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1832..1851 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1869..2324) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2006..2027) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2218..2237 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2351..2455 /gene="bla" /label=AmpR promoter promoter 2457..2814 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2480..2500) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2665..2800 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2727..2746 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2849..3643 /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(2903..2922) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3513..3532 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3831..3850) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3875..3922 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4251..4839 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4740..4759 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgagct caagcttcgg gccgcctgca gatcttcgtg 1381 aagactctga ctggtaagac catcaccctc gaggttgagc ccagtgacac cattgagaat 1441 gtcaaggcaa agatccaaga taaggaaggc atccctcctg accagcagag gctgatcttt 1501 gctggaaaac agctggaaga tgggcgcacc ctgtctgact acaacatcca gaaagagtcc 1561 accctgcacc tggtactccg tctcagaggt gggtgaggta ccgcgggccc gggatccacc 1621 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta 1681 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 1741 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 1801 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 1861 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 1921 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 1981 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 2041 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 2101 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 2161 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 2221 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 2281 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg 2341 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 2401 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga 2461 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 2521 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 2581 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2641 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 2701 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 2761 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga 2821 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 2881 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 2941 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 3001 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc 3061 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 3121 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 3181 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 3241 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 3301 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 3361 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 3421 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 3481 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 3541 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 3601 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 3661 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 3721 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 3781 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg 3841 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg 3901 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 3961 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 4021 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 4081 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt 4141 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 4201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 4261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 4321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 4381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 4441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 4501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 4561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 4621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 4681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 4741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 4801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 4861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 4921 gattctgtgg ataaccgtat taccgccatg cat //