LOCUS Exported 4996 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRK5-HA-Ubiquitin-K6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4996) AUTHORS Livingston CM, Ifrim MF, Cowan AE, Weller SK TITLE Virus-Induced Chaperone-Enriched (VICE) domains function as nuclear protein quality control centers during HSV-1 infection. JOURNAL PLoS Pathog. 2009 Oct . 5(10):e1000619. PUBMED 19816571 REFERENCE 2 (bases 1 to 4996) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4996 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 98..119 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 134..164 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 172..188 /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)." primer_bind 177..199 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 196..212 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 196..212 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 241..620 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 621..824 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 775..795 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 821..845 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1055..1073 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1055..1072 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 1086..1105 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1190..1199 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1202..1228 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1259..1486 /codon_start=1 /product="human ubiquitin variant containing Lys-6, with all other lysines mutated to arginine" /label=ubiquitin-K6 /translation="MQIFVKTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG" regulatory 1523..1532 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 1536..1670 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1573..1592) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1627..1646 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 1739..2096 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(1762..1782) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1947..2082 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2009..2028 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2116..2133) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2116..2132) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2125..2147) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2345..2800 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2432..2451) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2642..2663 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2817..2836) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2936..2958 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2996..3014) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3082..3186 /gene="bla" /label=AmpR promoter CDS 3187..4047 /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 complement(3405..3424) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4218..4806 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4707..4726 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4960..4977 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt 61 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt 121 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg 181 gataacaatt tcacacagga aacagctatg acatgattac gaattaattc gagctcgccc 241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 841 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat 901 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg 961 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca 1021 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat 1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat 1141 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatggg 1201 ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg ggtcgaccat 1261 gcagatcttc gtcaaaacgt taaccggtag aaccataact ctagaagttg aaccatccga 1321 taccatcgaa aacgttagag ctagaattca agacagagaa ggcattccac ctgatcaaca 1381 aagattgatc tttgccggta gacagctcga ggacggtaga acgctgtctg attacaacat 1441 tcagagagag tcgaccttac atcttgtctt aagactaaga ggtggttgag cggccgctaa 1501 gtaagtaagg atccccagct tggccgccat ggcccaactt gtttattgca gcttataatg 1561 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 1621 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat 1681 tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct 1741 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 1801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 1861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 1921 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 1981 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 2041 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 2101 tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 2161 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 2221 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat 2281 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca 2341 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2401 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 2461 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 2521 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt 2581 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 2641 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttaat 2701 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 2761 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca 2821 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg 2881 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 2941 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc 3001 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 3061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 3121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 3181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 3241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 3301 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 3361 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 3421 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 3481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 3541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 3601 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 3661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 3721 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 3781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 3841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 3901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 3961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 4021 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 4081 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 4141 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 4201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 4381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 4681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 4741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 4801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 4861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 4921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 4981 gcggaagagc gcccaa //