LOCUS Exported 7186 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS VCP(wt)-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7186) AUTHORS Tresse E, Salomons FA, Vesa J, Bott LC, Kimonis V, Yao TP, Dantuma NP, Taylor JP TITLE VCP/p97 is essential for maturation of ubiquitin-containing autophagosomes and this function is impaired by mutations that cause IBMPFD. JOURNAL Autophagy. 2010 Feb 22. 6(2):. PUBMED 20104022 REFERENCE 2 (bases 1 to 7186) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7186 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..204 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 155..175 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2706..2715 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2712..3425 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELY" CDS 2712..3425 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELY" primer_bind complement(2757..2778) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3018..3037) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3365..3386 /label=EGFP-C /note="EGFP, forward primer" CDS 3435..3452 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 3456..3482 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 3610..3731 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3647..3666) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3701..3720 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3738..4193) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3875..3896) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4087..4106 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4220..4324 /gene="bla" /label=AmpR promoter promoter 4326..4683 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4349..4369) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4534..4669 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4596..4615 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4718..5512 /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(4772..4791) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5382..5401 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5700..5719) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5744..5791 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6120..6708 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6609..6628 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 6883..7186 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg 241 actcagatct cgagctcaag cttgcgcgcg ccatggcttc tggagccgat tcaaaaggtg 301 atgacctatc aacagccatt ctcaaacaga agaaccgtcc caatcggtta attgttgatg 361 aagccatcaa tgaggacaac agtgtggtgt ccttgtccca gcccaagatg gatgaattgc 421 agttgttccg aggtgacaca gtgttgctga aaggaaagaa gagacgagaa gctgtttgca 481 tcgtcctttc tgatgatact tgttctgatg agaagattcg gatgaataga gttgttcgga 541 ataaccttcg tgtacgccta ggggatgtca tcagcatcca gccatgccct gatgtgaagt 601 acggcaaacg tatccatgtg ctgcccattg atgacacagt ggaaggcatt actggtaatc 661 tcttcgaggt ataccttaag ccgtacttcc tggaagcgta tcgacccatc cggaaaggag 721 acatttttct tgtccgtggt gggatgcgtg ctgtggagtt caaagtggtg gaaacagatc 781 ctagccctta ttgcattgtt gctccagaca cagtgatcca ctgcgaaggg gagcctatca 841 aacgagagga tgaggaagag tccttgaatg aagtagggta tgatgacatt ggtggctgca 901 ggaagcagct agctcagata aaggagatgg tggaactgcc cctgagacat cctgccctct 961 ttaaggcaat tggtgtgaag cctcctagag gaatcctgct ttacggacct cctggaacag 1021 gaaagaccct gattgctcga gctgtagcaa atgagactgg agccttcttc ttcttgatca 1081 atggtcctga gatcatgagc aaattggctg gtgagtctga gagcaacctt cgtaaagcct 1141 ttgaggaggc tgagaagaat gctcctgcca tcatcttcat tgatgagcta gatgccatcg 1201 ctcccaaaag agagaaaact catggcgagg tggagcggcg cattgtatca cagttgttga 1261 ccctcatgga tggcctaaag cagagggcac atgtgattgt tatggcagca accaacagac 1321 ccaacagcat tgacccagct ctacggcgat ttggtcgctt tgacagggag gtagatattg 1381 gaattcctga tgctacagga cgcttagaga ttcttcagat ccataccaag aacatgaagc 1441 tggcagatga tgtggacctg gaacaggtag ccaatgagac tcacgggcat gtgggtgctg 1501 acttagcagc cctgtgctca gaggctgctc tgcaagccat ccgcaagaag atggatctca 1561 ttgacctaga ggatgagacc attgatgccg aggtcatgaa ctctctagca gttactatgg 1621 atgacttccg gtgggccttg agccagagta acccatcagc actgcgggaa accgtggtag 1681 aggtgccaca ggtaacctgg gaagacatcg ggggcctaga ggatgtcaaa cgtgagctac 1741 aggagctggt ccagtatcct gtggagcacc cagacaaatt cctgaagttt ggcatgacac 1801 cttccaaggg agttctgttc tatggacctc ctggctgtgg gaaaactttg ttggccaaag 1861 ccattgctaa tgaatgccag gccaacttca tctccatcaa gggtcctgag ctgctcacca 1921 tgtggtttgg ggagtctgag gccaatgtca gagaaatctt tgacaaggcc cgccaagctg 1981 ccccctgtgt gctattcttt gatgagctgg attcgattgc caaggctcgt ggaggtaaca 2041 ttggagatgg tggtggggct gctgaccgag tcatcaacca gatcctgaca gaaatggatg 2101 gcatgtccac taaaaaaaat gtgttcatca ttggcgctac caaccggcct gacatcattg 2161 atcctgccat cctcagacct ggccgtcttg atcagctcat ctacatccca cttcctgatg 2221 agaagtcccg tgttgccatc ctcaaggcta acctgcgcaa gtccccagtt gccaaggatg 2281 tggacttgga gttcctggct aaaatgacta atggcttctc tggagctgac ctgacagaga 2341 tttgccagcg tgcttgcaag ctggccatcc gtgaatccat cgagagtgag attaggcgag 2401 aacgagagag gcagacaaac ccatcagcca tggaggtaga agaggatgat ccagtgcctg 2461 agatccgtcg agatcacttt gaagaagcca tgcgctttgc gcgccgttct gtcagtgaca 2521 atgacattcg gaagtatgag atgtttgccc agacccttca gcagagtcgg ggctttggca 2581 gcttcagatt cccttcaggg aaccagggtg gagctggccc cagtcagggc agtggaggcg 2641 gcacaggtgg cagtgtatac acagaagaca atgatgatga cctgtatggc tcggatccac 2701 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 2761 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 2821 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 2881 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 2941 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 3001 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 3061 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 3121 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca 3181 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 3241 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 3301 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg 3361 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 3421 tgtacagcgc tggccaccac catcaccatc acggataccc atacgatgtt ccagattacg 3481 ctagctgagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta 3541 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 3601 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 3661 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 3721 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt 3781 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc 3841 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt 3901 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact 3961 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg 4021 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag 4081 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac 4141 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg 4201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 4261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 4321 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc 4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 4441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 4561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 4621 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 4681 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 4741 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 4801 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 4861 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc 4921 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 4981 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 5041 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 5101 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 5161 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 5221 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca 5281 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 5341 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 5401 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 5461 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 5521 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 5581 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 5641 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact 5701 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag 5761 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg 5821 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt 5881 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg 5941 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat 6001 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 6061 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 6121 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 6181 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 6241 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 6301 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 6361 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 6421 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 6481 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 6541 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 6601 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 6661 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 6721 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 6781 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc 6841 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 6901 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 6961 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 7021 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga 7081 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt 7141 tcctacttgg cagtacatct acgtattagt catcgctatt accatg //