LOCUS Exported 7948 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFRT/FLAG/HA-DEST EIF2C2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7948) AUTHORS Landthaler M, Gaidatzis D, Rothballer A, Chen PY, Soll SJ, Dinic L, Ojo T, Hafner M, Zavolan M, Tuschl T TITLE Molecular characterization of human Argonaute-containing ribonucleoprotein complexes and their bound target mRNAs. JOURNAL RNA. 2008 Oct 31. ():. PUBMED 18978028 REFERENCE 2 (bases 1 to 7948) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7948 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 946..955 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 955..978 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 979..1005 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind 1021..1045 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 3067..3076 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(3744..3768) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3821..3862 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3900..3917) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3906..4130 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4263..4282) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 4414..4461 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 4469..5491 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 5621..5742 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5658..5677) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5712..5731 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5791..5807) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5791..5807) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5804..5826) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5815..5831 /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)." promoter complement(5839..5869) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5884..5905 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6022..6039) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6193..6781) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6273..6292) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6952..7812) /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 7575..7594 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7813..7917) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gagctcggat cgatccatcg atagtgtggt ggaattctga tcgccgccgc catggactac 961 aaggacgacg atgacaagta cccttatgac gtgcccgatt acgctttgaa ccagatatca 1021 acaagtttgt acaaaaaagc aggctccacc atgggaacca attcagtcga catgtactcg 1081 ggagccggct cccgagtaca tgtcgacgcg tgggccggcc cacgcgtcga ctggatccgg 1141 atgtactcgg gagccggccc cgcacttgca cctcctgcgc cgccgccccc catccaagga 1201 tatgccttca agcctccacc tagacccgac tttgggacct ccgggagaac aatcaaatta 1261 caggccaatt tcttcgaaat ggacatcccc aaaattgaca tctatcatta tgaattggat 1321 atcaagccag agaagtgccc gaggagagtt aacagggaaa tcgtggaaca catggtccag 1381 cactttaaaa cacagatctt tggggatcgg aagcccgtgt ttgacggcag gaagaatcta 1441 tacacagcca tgccccttcc gattgggagg gacaaggtgg agctggaggt cacgctgcca 1501 ggagaaggca aggatcgcat cttcaaggtg tccatcaagt gggtgtcctg cgtgagcttg 1561 caggcgttac acgatgcact ttcagggcgg ctgcccagcg tcccttttga gacgatccag 1621 gccctggacg tggtcatgag gcacttgcca tccatgaggt acacccccgt gggccgctcc 1681 ttcttcaccg cgtccgaagg ctgctctaac cctcttggcg ggggccgaga agtgtggttt 1741 ggcttccatc agtccgtccg gccttctctc tggaaaatga tgctgaatat tgatgtgtca 1801 gcaacagcgt tttacaaggc acagccagta atcgagtttg tttgtgaagt tttggatttt 1861 aaaagtattg aagaacaaca aaaacctctg acagattccc aaagggtaaa gtttaccaaa 1921 gaaattaaag gtctaaaggt ggagataacg cactgtgggc agatgaagag gaagtaccgt 1981 gtctgcaatg tgacccggcg gcccgccagt caccaaacat tcccgctgca gcaggagagc 2041 gggcagacgg tggagtgcac ggtggcccag tatttcaagg acaggcacaa gttggttctg 2101 cgctaccccc acctcccatg tttacaagtc ggacaggagc agaaacacac ctaccttccc 2161 ctggaggtct gtaacattgt ggcaggacaa agatgtatta aaaaattaac ggacaatcag 2221 acctcaacca tgatcagagc aactgctagg tcggcgcccg atcggcaaga agagattagc 2281 aaattgatgc gaagtgcaag tttcaacaca gatccatacg tccgtgaatt tggaatcatg 2341 gtcaaagatg agatgacaga cgtgactggg cgggtgctgc agccgccctc catcctctac 2401 gggggcagga ataaagctat tgcgacccct gtccagggcg tctgggacat gcggaacaag 2461 cagttccaca cgggcatcga gatcaaggtg tgggccattg cgtgcttcgc cccccagcgc 2521 cagtgcacgg aagtccatct gaagtccttc acagagcagc tcagaaagat ctcgagagac 2581 gctggcatgc ccatccaggg ccagccgtgc ttctgcaaat acgcgcaggg ggcggacagc 2641 gtggagccca tgttccggca cctgaagaac acgtatgcgg gcctgcagct ggtggtggtc 2701 atcctgcccg gcaagacgcc cgtgtacgcc gaggtcaagc gcgtgggaga cacggtgctg 2761 gggatggcca cgcagtgcgt gcagatgaag aacgtgcaga ggaccacgcc acagaccctg 2821 tccaaccttt gcctgaagat caacgtcaag ctgggaggcg tgaacaacat cctgctgccc 2881 cagggcaggc cgccggtgtt ccagcagccc gtcatctttc tgggagcaga cgtcactcac 2941 ccccccgccg gggatgggaa gaagccctcc attgccgccg tggtgggcag catggacgcc 3001 caccccaatc gctactgcgc caccgtgcgc gtgcagcagc accggcagga gatcatacaa 3061 gacctggccg ccatggtccg cgagctcctc atccagttct acaagtccac gcgcttcaag 3121 cccacccgca tcatcttcta ccgcgacggt gtctctgaag gccagttcca gcaggttctc 3181 caccacgagt tgctggccat ccgtgaggcc tgtatcaagc tagaaaaaga ctaccagccc 3241 gggatcacct tcatcgtggt gcagaagagg caccacaccc ggctcttctg cactgacaag 3301 aacgagcggg ttgggaaaag tggaaacatt ccagcaggca cgactgtgga cacgaaaatc 3361 acccacccca ccgagttcga cttctacctg tgtagtcacg ctggcatcca ggggacaagc 3421 aggccttcgc actatcacgt cctctgggac gacaatcgtt tctcctctga tgagctgcag 3481 atcctaacct accagctgtg tcacacctac gtgcgctgca cacgctccgt gtccatccca 3541 gcgccagcat actacgctca cctggtggcc ttccgggcca ggtaccacct ggtggataag 3601 gaacatgaca gtgctgaagg aagccatacc tctgggcaga gtaacgggcg agaccaccaa 3661 gcactggcca aggcggtcca ggttcaccaa gacactctgc gcaccatgta ctttgcttga 3721 gcggccgcac tcgagatatc tagacccagc tttcttgtac aaagtggttg atatccagca 3781 cagtggcggc cgctcaagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc 3841 tctcctcggt ctcgattcta cgctcgagtc tagagggccc gtttaaaccc gctgatcagc 3901 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3961 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 4021 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 4081 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 4141 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 4201 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 4261 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 4321 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4381 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc 4441 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga 4501 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga 4561 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag 4621 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct 4681 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc 4741 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct 4801 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg 4861 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg 4921 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc 4981 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg 5041 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac 5101 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat 5161 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag 5221 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga 5281 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg 5341 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag 5401 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg 5461 ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc 5521 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 5581 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 5641 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 5701 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 5761 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5821 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5881 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5941 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6001 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6061 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6121 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6181 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6241 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6301 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6361 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6421 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6481 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6541 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 6601 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 6661 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6721 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6781 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 6841 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 6901 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 6961 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7021 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7081 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7141 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7201 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7261 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7321 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7381 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7441 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7501 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7561 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 7621 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 7681 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7741 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7801 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 7861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 7921 atttccccga aaagtgccac ctgacgtc //