LOCUS Exported 7088 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS mTOR_2 shRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7088) AUTHORS Sarbassov DD, Guertin DA, Ali SM, Sabatini DM TITLE Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. JOURNAL Science 2005 Feb 18;307(5712):1098-101. PUBMED 15718470 REFERENCE 2 (bases 1 to 7088) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7088 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 55..76 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 91..121 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 129..145 /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 134..156 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 153..169 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 153..169 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 188..208 /label=T3 /note="T3 promoter, forward primer" promoter 190..208 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 236..462 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 463..643 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 690..815 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1308..1541 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1726..1770 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 2068..2308 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 2068..2088 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2239..2258 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 2417..2534 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2583..3093 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 3012..3030 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 3115..3714 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3115..3134) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3611..3631 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3842..4075 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4147..4268 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4184..4203) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4238..4257 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4308..4443 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4370..4389 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4463..4482) /label=T7 /note="T7 promoter, forward primer" promoter complement(4464..4482) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4492..4509) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4492..4508) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4501..4523) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4650..5105 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4737..4756) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4947..4968 /label=F1ori-F /note="F1 origin, forward primer" promoter 5131..5235 /gene="bla" /label=AmpR promoter CDS 5236..6096 /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(5454..5473) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6267..6855 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6756..6775 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7009..7026 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt 61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt 121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc 181 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag 241 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta 301 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc 361 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat 421 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac 481 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa 541 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag 601 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg 661 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag 721 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg 781 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat 841 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata 901 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat 961 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag 1021 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga 1081 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca 1141 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg 1201 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc 1261 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc 1321 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta 1381 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt 1441 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga 1501 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct 1561 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg 1621 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca 1681 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa 1741 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg 1801 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt 1861 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc 1921 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag 1981 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg atcacgagac 2041 tagcctcgag cggccgcccc cttcaccgag ggcctatttc ccatgattcc ttcatatttg 2101 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag 2161 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta 2221 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc 2281 ttggctttat atatcttgtg gaaaggacga aacaccggcc gcattgtctc tatcaagttc 2341 ttcctgtcaa acttgataga gacaatgcgg tttttgaatt ctcgacctcg agacaaatgg 2401 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa 2461 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa 2521 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg ccgcggctcg 2581 agggggttgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc 2641 tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg 2701 tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt 2761 cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa 2821 acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca 2881 gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc 2941 agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc 3001 ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc 3061 ctcgttgacc gaatcaccga cctctctccc cagggggatc caccggagct taccatgacc 3121 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 3181 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 3241 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 3301 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 3361 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 3421 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 3481 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 3541 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 3601 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 3661 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg 3721 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc ggtaccttta 3781 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 3841 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct 3901 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 3961 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 4021 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 4081 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 4141 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 4261 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa 4321 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 4381 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 4441 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt 4501 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc 4561 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca 4621 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc 4681 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 4741 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 4801 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 4861 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 4921 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 4981 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 5041 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac 5101 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 5161 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 5221 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 5281 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 5341 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 5401 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 5461 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 5521 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 5581 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 5641 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 5701 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 5761 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 5821 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 5881 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 5941 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 6001 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 6061 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 6121 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 6181 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 6241 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 6301 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 6361 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 6421 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 6481 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 6541 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 6601 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 6661 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 6721 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 6781 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 6841 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 6901 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 6961 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 7021 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 7081 cattaatg //