LOCUS Exported 4996 bp ds-DNA circular SYN 19-JAN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRK5-HA-Ubiquitin-K11 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 Jan 19, 2022 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 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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 186..208 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 205..221 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 250..629 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 630..833 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 784..804 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 830..854 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1064..1082 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1064..1081 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 1095..1114 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1199..1208 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1211..1237 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1268..1495 /codon_start=1 /product="human ubiquitin variant containing Lys-11, with all other lysines mutated to arginine" /label=ubiquitin-K11 /translation="MQIFVRTLTGKTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG" regulatory 1532..1541 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 1545..1679 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1582..1601) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1636..1655 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 1748..2105 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(1771..1791) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1956..2091 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2018..2037 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2125..2142) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2125..2141) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2134..2156) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2354..2809 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2441..2460) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2651..2672 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2826..2845) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2945..2967 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3005..3023) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3091..3195 /gene="bla" /label=AmpR promoter CDS 3196..4056 /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(3414..3433) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4227..4815 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4716..4735 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4969..4986 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg 241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag 301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct 361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc 421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg 481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat 541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag 841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc 901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca 961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac 1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata 1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct 1141 cggttctatc gattgaattc ccgaaccgac agtcggtctc ttcaccaagg ccattcgcgc 1201 caccatgggc tacccctatg atgtgcctga ctacgcagat ctcaatggtg gtggtggtgg 1261 gtcgaccatg cagatcttcg tcagaacgtt aaccggtaaa accataactc tagaagttga 1321 accatccgat accatcgaaa acgttagagc tagaattcaa gacagagaag gcattccacc 1381 tgatcaacaa agattgatct ttgccggtag acagctcgag gacggtagaa cgctgtctga 1441 ttacaacatt cagagagagt cgaccttaca tcttgtctta agactaagag gtggttgagc 1501 ggccgctaag taagtaagga tccccagctt ggccgccatg gcccaacttg tttattgcag 1561 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 1621 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg 1681 ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag 1741 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt 1801 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 1861 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 1921 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 1981 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 2041 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 2101 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa 2161 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 2221 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 2281 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca 2341 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2401 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2461 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2521 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt 2581 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 2641 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat 2701 tcttttaatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 2761 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact 2821 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 2881 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 2941 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 3001 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 3061 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 3121 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 3181 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 3241 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 3301 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 3361 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 3421 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 3481 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 3541 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 3601 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 3661 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 3721 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 3781 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 3841 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 3901 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 3961 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 4021 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 4081 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 4141 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 4201 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 4261 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 4321 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 4381 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 4441 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 4501 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 4561 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 4621 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 4681 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 4741 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 4801 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 4861 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 4921 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 4981 agcgaggaag cggaag //