LOCUS Exported 6427 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRK5-HA GST RagA 66L SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6427) 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 6427) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6427 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 581..605 /label=LNCX /note="Human CMV promoter, forward primer" promoter 815..833 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 815..832 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 846..865 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 950..959 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 962..988 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1019..1672 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 1630..1652 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" regulatory 2714..2723 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 2727..2861 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2764..2783) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2818..2837 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 2930..3287 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2953..2973) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3138..3273 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3200..3219 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3307..3324) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3307..3323) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3316..3338) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3536..3991 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3623..3642) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3833..3854 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4008..4027) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4127..4149 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4187..4205) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4273..4377 /gene="bla" /label=AmpR promoter CDS 4378..5238 /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(4596..4615) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5409..5997 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5898..5917 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6151..6168 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6285..6306 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6321..6351 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6359..6375 /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 6364..6386 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6383..6399 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6383..6399 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat 661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg 721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca 781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat 841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat 901 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatggg 961 ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg ggtcgagaat 1021 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt 1081 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg 1141 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg 1201 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat 1261 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga 1321 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga 1381 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac 1441 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt 1501 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa 1561 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg 1621 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aaggtggagg 1681 tggtgggtcg aggggaggtt ctggtggatc aggaggttca ggtggatctg ggtcgacgat 1741 gccaaataca gccatgaaga aaaaggtgct gctgatgggg aagagcgggt cggggaagac 1801 cagcatgagg tcgataatct tcgccaatta cattgctcgc gacacccggc gcctgggggc 1861 caccattgac gtggaacact cccacgtccg attcctaggg aacctggtgc tgaacctgtg 1921 ggactgtggc ggtctggaca ccttcatgga aaattacttc accagccagc gagacaatat 1981 cttccgtaac gtggaagttt tgatttacgt gtttgacgtg gagagccgcg aactggaaaa 2041 ggacatgcat tattaccagt cgtgtctgga ggccatcctc cagaactctc ctgacgccaa 2101 aatcttctgc ctggtgcaca aaatggatct ggttcaggag gatcagcgtg acctgatttt 2161 taaagagcga gaggaagacc tgaggcgtct gtctcgcccg ctggagtgtg cttgttttcg 2221 aacgtccatc tgggatgaga cgctctacaa agcctggtcc agcatcgtct accagctgat 2281 tcccaacgtt cagcagctgg agatgaacct caggaatttt gcccaaatca ttgaggccga 2341 tgaagttctg ctgttcgaaa gagctacatt cttggttatt tcccactacc agtgcaaaga 2401 gcagcgcgac gtccaccggt ttgagaagat cagcaacatc atcaaacagt tcaagctgag 2461 ctgcagtaaa ttggccgctt ccttccagag catggaagtt aggaattcca acttcgctgc 2521 tttcatcgac atcttcacct caaatacgta cgtgatggtg gtcatgtcag atccgtcgat 2581 cccttctgcg gccactctga tcaacattcg caatgcccgg aaacactttg agaagctgga 2641 gagagtggat ggccccaagc acagtctcct tatgcgttga gcggccgcta agtaagtaag 2701 gatccccagc ttggccgcca tggcccaact tgtttattgc agcttataat ggttacaaat 2761 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 2821 gtttgtccaa actcatcaat gtatcttatc atgtctggat cgggaattaa ttcggcgcag 2881 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa 2941 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctgttaacag 3301 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact 3361 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac 3421 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt 3481 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg 3541 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3601 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3661 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 3721 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg 3781 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3841 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg 3901 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3961 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct 4021 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 4081 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 4141 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 4201 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 4261 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 4321 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 4381 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 4441 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 4501 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 4561 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 4621 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 4681 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 4741 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 4801 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 4861 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 4921 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 4981 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 5041 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 5101 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 5161 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 5221 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 5281 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 5341 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 5401 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 5461 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 5521 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 5581 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 5641 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 5701 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 5761 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 5821 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 5881 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 5941 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 6001 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 6061 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 6121 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 6181 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 6241 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 6301 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 6361 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttaattcgag 6421 ctcgccc //