LOCUS Exported 6688 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRK5-HA GST RagD 121L SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6688) AUTHORS Sancak Y, Peterson TR, Shaul YD, Lindquist RA, Thoreen CC, Bar-Peled L, Sabatini DM TITLE The Rag GTPases bind raptor and mediate amino acid signaling to mTORC1. JOURNAL Science. 2008 Jun 13. 320(5882):1496-501. PUBMED 18497260 REFERENCE 2 (bases 1 to 6688) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6688 /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 975..1001 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1032..1685 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 1643..1665 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" regulatory 2988..2997 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 3001..3135 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3038..3057) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3092..3111 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 3204..3561 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3227..3247) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3412..3547 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3474..3493 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3581..3598) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3581..3597) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3590..3612) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3810..4265 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3897..3916) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4107..4128 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4282..4301) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4401..4423 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4461..4479) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4547..4651 /gene="bla" /label=AmpR promoter CDS 4652..5512 /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(4870..4889) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5683..6271 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6172..6191 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6425..6442 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6559..6580 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6595..6625 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6633..6649 /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 6638..6660 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6657..6673 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6657..6673 /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 gggctacccc tatgatgtgc ctgactacgc agatctcaat ggtggtggtg 1021 gtgggtcgag aatgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca 1081 ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg 1141 aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt 1201 attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg 1261 acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag 1321 gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa 1381 ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt 1441 tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg 1501 acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag 1561 tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca 1621 agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc 1681 caaaaggtgg aggtggtggg tcgaggggag gttctggtgg atcaggaggt tcaggtggat 1741 ctgggtcgac gatgagccag gtgctgggga agccgcagcc gcaggacgag gacgacgcgg 1801 aggaggagga ggaggaggat gagctggtgg ggctagcgga ctacggagac gggcccgact 1861 cctccgacgc cgatccggac agcggcacag aggagggagt tctggacttc agtgacccct 1921 tcagcactga agtgaagccg agaatcctgc tcatgggcct gaggagaagc ggcaagtcgt 1981 ctattcagaa agttgtcttt cacaaaatgt ctcccaacga aactctgttc ttggagagca 2041 ctaataagat atgccgggaa gatgtttcca acagctcctt tgtcaatttt cagatttggg 2101 acttcccagg actgattgac ttttttgacc ctacatttga ctatgagatg atcttccggg 2161 gaacaggagc actgatattt gtcattgact cacaggatga ttacatggaa gccctggcca 2221 ggctccacct cacggtgacc agggcctaca aagtgaatac tgacatcaac ttcgaggtgt 2281 ttattcataa agtggatggt ctgtcagatg accacaaaat tgaaacccaa agagatattc 2341 accagagggc aaacgatgac cttgcagatg ctggattaga aaaaattcac ctcagctttt 2401 atctgacaag catatatgat cattcaatat ttgaagcttt tagcaaagtt gttcagaaac 2461 tgattccaca actcccaact ctggagaatt tgctgaacat ctttatctca aattctggaa 2521 ttgaaaaggc atttctattt gatgtggtca gtaaaattta tattgcaact gatagtactc 2581 cggtggatat gcaaacctat gagctctgct gtgatatgat agatgtggtt attgacatct 2641 cttgtattta tggtctcaaa gaagatggag caggaacccc ctatgacaag gaatccacag 2701 ccatcataaa gcttaataat acaaccgtgc tttatttaaa agaggtgaca aagttcctgg 2761 ctctcgtttg ctttgtcaga gaggaaagct ttgaaagaaa agggctaatt gactataatt 2821 ttcattgctt ccggaaggcc attcatgaag tttttgaggt gagaatgaaa gtagtaaaat 2881 ctcgaaaggt tcagaatcgg ctgcagaaga aaaagagagc cacccctaat gggaccccta 2941 gagtgctgct gtaggcggcc gctaagtaag taaggatccc cagcttggcc gccatggccc 3001 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 3061 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 3121 tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat aacctctgaa 3181 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag 3241 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 3301 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3361 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 3421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 3481 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 3541 ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc gttttacaac 3601 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 3661 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 3721 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt 3781 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc 3841 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 3901 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 3961 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 4021 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 4081 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 4141 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 4201 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac 4261 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg 4321 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta 4381 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc 4441 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat 4501 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat 4561 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata 4621 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct 4681 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa 4741 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa 4801 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt 4861 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg 4921 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca 4981 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa 5041 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt 5101 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc 5161 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa 5221 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga 5281 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc 5341 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga 5401 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga 5461 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga 5521 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 5581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 5641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 5701 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 5761 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 5821 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 5881 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 5941 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 6001 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 6061 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 6121 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 6181 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 6241 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 6301 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 6361 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 6421 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc 6481 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg 6541 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca 6601 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg 6661 aaacagctat gacatgatta cgaattaa //