LOCUS Exported 5185 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION N-terminal myc-tagged protein expression in mammaiian cells. ACCESSION . VERSION . KEYWORDS myc-POMP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5185) AUTHORS Yun YS, Kim KH, Tschida B, Sachs Z, Noble-Orcutt KE, Moriarity BS, Ai T, Ding R, Williams J, Chen L, Largaespada D, Kim DH TITLE mTORC1 Coordinates Protein Synthesis and Immunoproteasome Formation via PRAS40 to Prevent Accumulation of Protein Stress. JOURNAL Mol Cell. 2016 Feb 18;61(4):625-39. doi: 10.1016/j.molcel.2016.01.013. Epub 2016 Feb 11. PUBMED 26876939 REFERENCE 2 (bases 1 to 5185) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5185 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 14..393 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 394..597 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 548..568 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 594..618 /label=LNCX /note="Human CMV promoter, forward primer" promoter 828..846 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 828..845 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 859..878 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 963..972 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 972..1001 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" regulatory 1485..1494 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 1498..1632 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1535..1554) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1589..1608 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 1701..2058 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(1724..1744) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1909..2044 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1971..1990 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2078..2095) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2078..2094) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2087..2109) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2307..2762 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2394..2413) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2604..2625 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2779..2798) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2898..2920 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2958..2976) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3044..3148 /gene="bla" /label=AmpR promoter CDS 3149..4009 /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(3367..3386) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4180..4768 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4669..4688 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4922..4939 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5056..5077 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5092..5122 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5130..5146 /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 5135..5157 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5154..5170 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5154..5170 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca 61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt 601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga 661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt 721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta 781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac 841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc 901 acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc aaggccattc 961 gcgccaccat ggagcaaaag ctcatttctg aggaagatct caatggtggt ggtggtgggt 1021 cgaccatgaa tgccagagga cttggatctg agctaaagga cagtattcca gttactgaac 1081 tttcagcaag tggacctttt gaaagtcatg atcttcttcg gaaaggtttt tcttgtgtga 1141 aaaatgaact tttgcctagt catccccttg aattatcaga aaaaaatttc cagctcaacc 1201 aagataaaat gaatttttcc acactgagaa acattcaggg tctatttgct ccgctaaaat 1261 tacagatgga attcaaggca gtgcagcagg ttcagcgtct tccatttctt tcaagctcaa 1321 atctttcact ggatgttttg aggggtaatg atgagactat tggatttgag gatattctta 1381 atgatccatc acaaagcgaa gtcatgggag agccacactt gatggtggaa tataaacttg 1441 gtttactgta agcggccgct aagtaagtaa ggatccccag cttggccgcc atggcccaac 1501 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 1561 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 1621 catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga 1681 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta 1741 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 1801 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 1861 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 1921 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 1981 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 2041 ggcctaggct tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc 2101 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 2161 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 2221 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 2281 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg 2341 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 2401 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 2461 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 2521 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 2581 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 2641 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 2701 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat 2761 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca 2821 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag 2881 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa 2941 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 3001 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 3061 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 3121 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 3181 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 3241 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 3301 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 3361 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 3421 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 3481 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 3541 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 3601 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 3661 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 3721 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 3781 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 3841 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 3901 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 3961 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 4021 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 4081 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 4141 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 4201 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 4261 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 4321 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 4381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 4441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 4501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 4561 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 4621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 4681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 4741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 4801 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 4861 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 4921 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc 4981 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 5041 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt 5101 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa 5161 cagctatgac atgattacga attaa //