LOCUS Exported 7401 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION tRNA synthetase/tRNA pair for the incorporation of unnatural amino acid into proteins in mammalian cells in response to the amber codon TAG.. ACCESSION . VERSION . KEYWORDS Mm-PylRS-AF/Pyl-tRNACUA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7401) AUTHORS Spence JS, He R, Hoffmann HH, Das T, Thinon E, Rice CM, Peng T, Chandran K, Hang HC TITLE IFITM3 directly engages and shuttles incoming virus particles to lysosomes. JOURNAL Nat Chem Biol. 2019 Jan 14. pii: 10.1038/s41589-018-0213-2. doi: 10.1038/s41589-018-0213-2. PUBMED 30643282 REFERENCE 2 (bases 1 to 7401) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7401 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 46..425 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 426..629 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 580..600 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 626..650 /label=LNCX /note="Human CMV promoter, forward primer" promoter 860..878 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 860..877 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 891..910 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 962..971 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 971..994 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(2385..2402) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2391..2615 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2661..3089 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2748..2767) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2958..2979 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3098..3118) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3103..3432 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3283..3418 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3345..3364 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3481..4506 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 4636..4757 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4673..4692) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4727..4746 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4806..4822) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4806..4822) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4819..4841) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4830..4846 /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(4854..4884) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4899..4920 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5037..5054) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5208..5796) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5288..5307) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5967..6827) /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 6590..6609 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6828..6932) /gene="bla" /label=AmpR promoter promoter complement(7030..7270) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind complement(7080..7099) /label=LKO.1 5' /note="Human U6 promoter, forward primer" primer_bind complement(7250..7270) /label=hU6-F /note="Human U6 promoter, forward primer" ORIGIN 1 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga 61 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 121 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 301 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 361 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 421 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg 481 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 541 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg 601 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac 661 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc 721 gggaacggtg cattggaacg cggattcccc gtgccaagag tgacgtaagt accgcctata 781 gagtctatag gcccaccccc ttggcttcgt tagaacgcgg ctacaattaa tacataacct 841 tatgtatcat acacatacga tttaggtgac actatagaat aacatccact ttgcctttct 901 ctccacaggt gtccactccc aggtccaact gcacgggcta gcgtttaaac ttaagaagct 961 tgccaccatg gactacaagg acgacgacga caagatggac aagaagcccc tgaacaccct 1021 gatcagcgcc acaggactgt ggatgtccag aaccggcacc atccacaaga tcaagcacca 1081 cgaggtgtcc cggtccaaaa tctacatcga gatggcctgc ggcgatcacc tggtcgtcaa 1141 caacagcaga agcagccgga cagccagagc cctgcggcac cacaagtaca gaaagacctg 1201 caagcggtgc agagtgtccg acgaggacct gaacaagttc ctgaccaagg ccaacgagga 1261 ccagaccagc gtgaaagtga aggtggtgtc cgcccccacc cggaccaaga aagccatgcc 1321 caagagcgtg gccagagccc ccaagcccct ggaaaacacc gaagccgctc aggcccagcc 1381 cagcggcagc aagttcagcc ccgccatccc cgtgtctacc caggaaagcg tcagcgtccc 1441 cgccagcgtg tccaccagca tctctagcat ctcaaccggc gccacagctt ctgccctggt 1501 caagggcaac accaacccca tcaccagcat gtctgcccct gtgcaggcct ctgccccagc 1561 cctgaccaag tcccagaccg accggctgga agtgctcctg aaccccaagg acgagatcag 1621 cctgaacagc ggcaagccct tccgggagct ggaaagcgag ctgctgagcc ggcggaagaa 1681 ggacctccag caaatctacg ccgaggaacg ggagaactac ctgggcaagc tggaaagaga 1741 gatcacccgg ttcttcgtgg accggggctt cctggaaatc aagagcccca tcctgatccc 1801 cctggagtac atcgagcgga tgggcatcga caacgacacc gagctgagca agcagatttt 1861 ccgggtggac aagaacttct gcctgcggcc catgctggcc cccaacctgg ccaactacct 1921 gcggaaactg gatcgcgctc tgcccgaccc catcaagatt ttcgagatcg gcccctgcta 1981 ccggaaagag agcgacggca aagagcacct ggaagagttt acaatgctga acttttgcca 2041 gatgggcagc ggctgcacca gagagaacct ggaatccatc atcaccgact ttctgaacca 2101 cctggggatc gacttcaaga tcgtgggcga cagctgcatg gtgttcggcg acaccctgga 2161 cgtgatgcac ggcgacctgg aactgtctag cgccgtcgtg ggacccatcc ctctggaccg 2221 ggagtggggc atcgataagc cctggatcgg agccggcttc ggcctggaac ggctgctgaa 2281 agtcaagcac gactttaaga acatcaagcg ggctgccaga agcgagagct actacaacgg 2341 catcagcacc aacctgtgat gataaggatc cacccgctga tcagcctcga ctgtgccttc 2401 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 2461 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2521 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2581 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2641 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2701 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2761 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2821 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2881 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 2941 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 3001 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 3061 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 3121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3181 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3361 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3421 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg 3481 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac 3541 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat 3601 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat 3661 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt 3721 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg 3781 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat 3841 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga 3901 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat 3961 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag 4021 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc 4081 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg 4141 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct 4201 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg 4261 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac 4321 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga 4381 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc 4441 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag 4501 gaatagcacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4561 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4621 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4681 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4741 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4801 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4861 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4921 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaaat ggcggaaacc 6961 ccgggaatct aacccggctg aacggattta gagtccattc gatctacatg atcaggtttc 7021 cggtgtttcg tcctttccac aagatatata aagccaagaa atcgaaatac tttcaagtta 7081 cggtaagcat atgatagtcc attttaaaac ataattttaa aactgcaaac tacccaagaa 7141 attattactt tctacgtcac gtattttgta ctaatatctt tgtgtttaca gtcaaattaa 7201 ttctaattat ctctctaaca gccttgtatc gtatatgcaa atatgaagga atcatgggaa 7261 ataggccctc ttcctgcccg accttcgcga tgtacgggcc agatatacgc caatattggc 7321 cattagccat attattcatt ggttatatag cataaatcaa tattggctat tggccattgc 7381 atacgttgta tctatatcat a //