LOCUS Exported 6319 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AGO1 knockdown. ACCESSION . VERSION . KEYWORDS pAPM-D4 miR30-AGO1 ts3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6319) AUTHORS Yurkovetskiy L, Guney MH, Kim K, Goh SL, McCauley S, Dauphin A, Diehl WE, Luban J TITLE Primate immunodeficiency virus proteins Vpx and Vpr counteract transcriptional repression of proviruses by the HUSH complex. JOURNAL Nat Microbiol. 2018 Dec;3(12):1354-1361. doi: 10.1038/s41564-018-0256-x. Epub 2018 Oct 8. PUBMED 30297740 REFERENCE 2 (bases 1 to 6319) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6319 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(64..84) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 69..398 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 249..384 /label=SV40 ori /note="SV40 origin of replication" primer_bind 311..330 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" enhancer 410..789 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 791..989 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 944..964 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 1007..1187 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1234..1359 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1856..2089 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2274..2318 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2616..2733 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2870..3277 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" ncRNA 3992..4006 /label=miR-30a loop /note="loop from the the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" LTR 4140..4320 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(4391..4979) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4471..4490) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5150..6010) /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 5773..5792 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6011..6115) /gene="bla" /label=AmpR promoter primer_bind complement(6190..6209) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc 61 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 121 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 241 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 301 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 361 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgcgttg acattgatta 421 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 481 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 541 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 601 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 661 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 721 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 781 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 841 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 901 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 961 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt 1021 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 1081 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 1141 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac 1201 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct 1261 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact 1321 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt 1381 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa 1441 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa 1501 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca 1561 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata 1621 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag 1681 accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg 1741 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1801 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1861 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc 1921 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1981 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 2041 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 2101 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 2161 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca 2221 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg 2281 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa 2341 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa 2401 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt 2461 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag 2521 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc 2581 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg 2641 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa 2701 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat 2761 ccagtttggt taattaactg cagccccgat aaaataaaag attttattta gtctccagaa 2821 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagctgcag taacgccatt 2881 ttgcaaggca tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac 2941 atgaaaatag ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc 3001 ccggggccaa gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat 3061 ggtccccaga tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc 3121 tcccccaagg acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct 3181 cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact 3241 cggcgcgcca gtcctccgac agactgagtc gcccgggggt ctagaacgcg tgccgccatg 3301 accgaataca aacctaccgt gaggctggct acaagagatg atgtcccaag ggctgtgaga 3361 acactggccg ccgcttttgc cgattaccct gccacacgcc acactgtgga cccagatcgg 3421 catatcgaga gagtgactga gctgcaggaa ctgttcctga cccgagtggg cctggacatt 3481 gggaaggtct gggtcgcaga cgatggagca gctgtggctg tctggaccac accagagagc 3541 gtggaagccg gagctgtctt tgcagagatc ggccctagaa tggcagaact gagcggctcc 3601 aggctggcag cacagcagca gatggaggga ctgctggccc cacacaggcc taaggaacca 3661 gcatggttcc tggctaccgt gggggtctct cctgaccatc agggcaaagg actgggaagt 3721 gctgtggtcc tgccaggagt ggaggctgca gaacgagctg gagtccctgc atttctggag 3781 acctctgctc cacgaaacct gcccttctat gaacggctgg gctttactgt gaccgcagat 3841 gtggaggtcc ccgaaggacc taggacctgg tgcatgacac gcaaacccgg cgcctgagcg 3901 atcgccgcgg ccgccttctt aacccaacag aaggctcgag aaggtatatt gctgttgaca 3961 gtgagcgatg cctcaagatc aatgtcaaat agtgaagcca cagatgtatt tgacattgat 4021 cttgaggcag tgcctactgc ctcggacttc aaggggctag aattcggcag ctgtagatct 4081 tagccacttt ttaaaagaaa aggggggact ggaagggcta actgcatccg gactgtactg 4141 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 4201 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 4261 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 4321 gggcccgttt catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 4381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 4441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 4501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 4561 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 4621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 4681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 4741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 4801 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 4861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 4921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 4981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5281 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 5401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 5461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 5521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 5581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 5641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 5701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 5761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 5821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 5881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 5941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6121 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg 6181 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg 6241 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt 6301 gaccgacaat tgcatgaag //