LOCUS Exported 9295 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses lambdaN-HA-tagged XRN1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pLNHA-C1-HsXRN1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9295) AUTHORS Braun JE, Truffault V, Boland A, Huntzinger E, Chang CT, Haas G, Weichenrieder O, Coles M, Izaurralde E TITLE A direct interaction between DCP1 and XRN1 couples mRNA decapping to 5' exonucleolytic degradation. JOURNAL Nat Struct Mol Biol. 2012 Dec;19(12):1324-31. doi: 10.1038/nsmb.2413. Epub 2012 Nov 11. PUBMED 23142987 REFERENCE 2 (bases 1 to 9295) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9295 /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" primer_bind 613..632 /label=T7 /note="T7 promoter, forward primer" promoter 613..631 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 659..668 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 665..730 /codon_start=1 /product="N-terminal RNA-binding domain from the lambda bacteriophage antiterminator protein N (Baron-Benhamou et al., 2004)" /label=-lambda N peptide /translation="MDAQTRRRERRAEKQAQWKAAN" CDS 755..781 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 6083..6204 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6120..6139) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6174..6193 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(6211..6666) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6348..6369) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6560..6579 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6693..6797 /gene="bla" /label=AmpR promoter promoter 6799..7156 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6822..6842) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 7007..7142 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7069..7088 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7191..7985 /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(7245..7264) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7855..7874 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(8173..8192) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 8217..8264 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 8593..9181 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9082..9101 /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 gcagagctct ctggctaact agagaaccca ctgcttactg 601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc 661 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa 721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc 781 tcctccatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcatgggagt 841 ccccaagttt tacagatgga tctcagagcg gtatccctgt ctcagcgaag tggtgaaaga 901 gcatcagatt cctgaatttg acaacttgta cctggatatg aatggaatta tacatcagtg 961 ctcccatcct aatgatgatg atgttcactt tagaatttca gatgataaaa tctttactga 1021 tatttttcac tacctggagg tgttgtttcg cattattaaa cccaggaaag tgttctttat 1081 ggctgtagat ggtgtggctc ctcgagcaaa aatgaaccag cagcgtggga ggcgttttag 1141 gtcagcaaag gaggcagaag acaaaattaa aaaggcaata gagaagggag aaactcttcc 1201 tacagaggcc agatttgatt ccaactgtat cacaccagga actgaattta tggccaggtt 1261 acatgaacat ctgaagtatt ttgtaaatat gaaaatttcc acagacaagt catggcaagg 1321 agttaccatc tacttctcag gccatgagac tcctggagaa ggagagcata aaatcatgga 1381 atttatcaga tccgagaaag caaagccaga tcatgatcca aacaccagac actgtcttta 1441 tggtttagat gctgacttga ttatgcttgg attaacaagt catgaggcac atttttctct 1501 cttaagagaa gaagttcgat ttggtggcaa aaaaacacaa cgggtatgtg ctccagaaga 1561 aactacattt caccttctac acttgtcttt aatgagagag tatattgact atgagttttc 1621 agtattaaaa gaaaagatca catttaaata tgatattgaa aggataatag atgattggat 1681 tttgatgggg tttcttgttg gtaatgattt tatccctcat ctacctcatt tacatattaa 1741 tcatgatgca ctgcctcttc tttatggaac atatgttacc atcctgccag aacttggggg 1801 ttatattaat gaaagtgggc acctcaactt acctcgattt gagaaatacc ttgtgaaact 1861 atcagatttt gatcgggagc acttcagtga agtttttgtg gacctaaaat ggtttgaaag 1921 caaagttggt aacaagtacc tcaatgaagc agcaggtgtc gcagcagaag aagccaggaa 1981 ctacaaggaa aagaaaaagt taaagggcca ggaaaattct ctgtgttgga ctgctttaga 2041 caaaaatgaa ggcgaaatga taacttctaa ggataattta gaagatgaga ctgaagatga 2101 tgacctattt gaaactgagt ttagacaata taaaagaaca tattacatga cgaagatggg 2161 ggttgacgta gtatctgatg actttctggc tgatcaagct gcatgttatg ttcaggcaat 2221 acagtggatt ttgcactatt actatcatgg agttcagtcc tggagctggt attatcctta 2281 tcattatgcg cctttcctgt ctgatataca caacatcagt acactcaaaa tccattttga 2341 actaggaaaa ccttttaagc catttgaaca gcttcttgct gtacttccag cagccagcaa 2401 aaatttactt cctgcatgct accagcattt gatgaccaat gaagactcac caattataga 2461 atattaccca cctgatttta aaactgacct aaatgggaaa caacaggaat gggaagctgt 2521 ggtgttaatc ccttttattg atgagaagcg attattggaa gccatggaga catgtaacca 2581 ctccctcaaa aaggaagaga ggaaaagaaa ccaacatagt gagtgcctaa tgtgctggta 2641 tgatagagac acagagttta tctatccttc tccatggcca gaaaagttcc ctgccataga 2701 acgatgttgt acaaggtata aaataatatc cttagatgct tggcgtgtag acataaacaa 2761 aaacaaaata accagaattg accagaaagc attatatttc tgtggatttc ctactctgaa 2821 acacatcaga cacaaatttt ttttgaagaa aagtggtgtt caagtattcc agcaaagcag 2881 tcgtggagaa aacatgatgt tggaaatctt agtggatgca gaatcagatg aacttaccgt 2941 agaaaatgta gcttcatcag tgcttggaaa atctgtcttt gttaattggc ctcaccttga 3001 ggaagctaga gtcgtggctg tatcagatgg agaaactaag ttttacttgg aagaacctcc 3061 aggaacacag aagctttatt caggaagaac tgccccacca tctaaagtgg ttcatcttgg 3121 agataaagaa caatctaact gggcaaaaga agtacaagga atttcagaac actacctgag 3181 aagaaaagga ataataataa atgaaacatc tgcagttgtg tatgctcagt tactcacagg 3241 tcgtaaatat caaataaatc aaaatggtga agttcgtcta gagaaacagt ggtcaaaaca 3301 agttgttcct tttgtttatc aaactattgt caaggacatc cgagctttcg actcccgttt 3361 ctccaatatc aaaacattgg atgatttgtt tcctctgaga agtatggtct ttatgctggg 3421 aactccctat tatggctgca ctggagaagt tcaggattca ggtgatgtga ttacagaagg 3481 taggattcgt gtgattttca gcattccatg tgaacccaat cttgatgctt taatacagaa 3541 ccagcataaa tattctataa agtacaaccc aggatatgtg ttggccagtc gccttggagt 3601 gagtggatac cttgtttcaa ggtttacagg aagtattttt attggaagag gatctaggag 3661 aaaccctcat ggagaccata aagcaaatgt gggtttaaat ctcaaattca acaagaaaaa 3721 tgaggaggta cctggatata ctaagaaagt tggaagtgaa tggatgtatt catctgcagc 3781 agaacaactt ctggcagagt acttagagag agctccagaa ctatttagtt atatagccaa 3841 aaatagccaa gaggatgtgt tctatgaaga tgacatttgg cctggagaaa atgagaatgg 3901 tgctgaaaaa gttcaagaaa ttattacttg gctaaaagga catcctgtca gtactttatc 3961 tcgttcttct tgtgatttac aaattctgga tgcagctatt gttgagaaaa ttgaggaaga 4021 agtcgaaaag tgcaagcaaa gaaagaataa taagaaggtg cgagtaacag tgaaacccca 4081 tttgctatac agacctttag aacagcaaca tggagtcatt cctgatcggg atgcagaatt 4141 ttgtcttttt gaccgtgttg taaatgtgag agaaaacttc tcagttccag ttggccttcg 4201 aggcaccatc ataggaataa aaggagctaa tagagaagcc gatgtactat ttgaagtatt 4261 atttgatgaa gaatttcctg gagggttaac aataagatgc tcacctggta gaggttatcg 4321 actgccaaca agtgccttgg tgaacctttc tcatgggagt cgctctgaaa ctggaaatca 4381 gaagttgaca gccatcgtaa aaccacaacc agctgtacat caacatagct caagttcatc 4441 agtttcctct gggcatttgg gagccctcaa ccattcccct caatcacttt ttgttcctac 4501 tcaagtacct actaaagatg atgatgaatt ctgcaacatt tggcagtcct tacagggatc 4561 tggaaagatg caatactttc agccaactat acaagagaag ggtgcagttc tacctcaaga 4621 aataagccaa gtaaatcaac atcataaatc tggctttaat gacaacagtg ttaaatatca 4681 gcaaagaaaa catgaccctc acagaaaatt taaagaagag tgtaagagtc ctaaagctga 4741 gtgttggtcc caaaaaatgt ccaataagca gcctaactct ggaattgaga actttttagc 4801 atctttgaat atctccaaag aaaatgaagt acagtcatct catcatgggg agcctccaag 4861 tgaagagcat ttgtcaccac agtcatttgc catgggaaca cggatgctta aagaaattct 4921 aaaaattgat ggctctaaca ctgtggacca taagaatgaa atcaaacaga ttgctaatga 4981 aatccctgtt tcctctaaca gaagagatga atatggatta ccctctcagc ctaaacaaaa 5041 taagaaatta gcatcttata tgaacaagcc tcacagtgct aatgagtacc ataatgttca 5101 gtctatggac aatatgtgtt ggcctgcccc cagccagatc cctcctgtat ccacaccagt 5161 aactgaactt tctcgaattt gttcccttgt tggaatgcca caacctgatt tctcctttct 5221 taggatgcca cagacaatga ccgtttgcca agtaaaatta tctaatggct tactggtaca 5281 tgggccacag tgccactctg aaaatgaagc caaagagaaa gctgcacttt ttgctttaca 5341 acagttgggc tccttaggca tgaatttccc tttgccttca caagtatttg caaattatcc 5401 ttcagctgta ccacctggaa ccattcctcc agcctttccc ccacctactg gctgggatca 5461 ctatggaagc aactatgcat tgggggcagc taatataatg ccttcgtcgt ctcatctctt 5521 tggctcaatg ccatggggac catcggtgcc agttcctggg aagcccttcc atcatacttt 5581 atattctggg accatgccca tggctggggg aataccaggg ggtgtgcaca atcagtttat 5641 acctctgcag gttactaaaa aaagggttgc aaacaaaaag aactttgaga ataaggaagc 5701 ccagagttct caagccactc cagttcagac tagccagcca gattcttcca acattgtcaa 5761 agtaagtcca cgggagagct catcagcttc tttgaagtcc tctccgattg ctcaacctgc 5821 atcttctttt caagttgaaa ctgcctctca aggccatagt atatctcacc ataagtcaac 5881 accaatctct tcttcaagaa gaaaatcaag aaaactggct gttaattttg gtgtttctaa 5941 accttctgag taaggatcca ccggatctag ataactgatc ataatcagcc ataccacatt 6001 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa 6061 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag 6121 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 6181 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat 6241 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 6301 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 6361 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 6421 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta 6481 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 6541 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 6601 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg 6661 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 6721 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 6781 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg 6841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 6901 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 6961 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 7021 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 7081 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 7141 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac 7201 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 7261 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 7321 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg 7381 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg 7441 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 7501 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 7561 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 7621 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 7681 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc 7741 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 7801 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 7861 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 7921 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 7981 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 8041 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 8101 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag 8161 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc 8221 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg 8281 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 8341 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 8401 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga 8461 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 8521 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 8581 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 8641 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 8701 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt 8761 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 8821 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 8881 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 8941 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 9001 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 9061 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 9121 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 9181 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 9241 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat //