LOCUS Exported 7084 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Rictor_2 shRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7084) 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 7084) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7084 /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 2413..2530 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2579..3089 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 3008..3026 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 3111..3710 /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(3111..3130) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3607..3627 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3838..4071 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4143..4264 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4180..4199) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4234..4253 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4304..4439 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4366..4385 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4459..4478) /label=T7 /note="T7 promoter, forward primer" promoter complement(4460..4478) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4488..4505) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4488..4504) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4497..4519) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4646..5101 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4733..4752) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4943..4964 /label=F1ori-F /note="F1 origin, forward primer" promoter 5127..5231 /gene="bla" /label=AmpR promoter CDS 5232..6092 /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(5450..5469) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6263..6851 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6752..6771 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7005..7022 /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 aacaccgggc agccttgaac tgtttaactt 2341 cctgtcatta aacagttcaa ggctgctttt tgaattctcg acctcgagac aaatggcagt 2401 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat 2461 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat 2521 tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggccgc ggctcgaggg 2581 ggttggggtt gcgccttttc caaggcagcc ctgggtttgc gcagggacgc ggctgctctg 2641 ggcgtggttc cgggaaacgc agcggcgccg accctgggtc tcgcacattc ttcacgtccg 2701 ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg ccccccggcg acgcttcctg 2761 ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg 2821 aagccgcacg tctcactagt accctcgcag acggacagcg ccagggagca atggcagcgc 2881 gccgaccgcg atgggctgtg gccaatagcg gctgctcagc agggcgcgcc gagagcagcg 2941 gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc 3001 ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg 3061 ttgaccgaat caccgacctc tctccccagg gggatccacc ggagcttacc atgaccgagt 3121 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg 3181 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg 3241 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 3301 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 3361 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 3421 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 3481 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 3541 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 3601 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 3661 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc 3721 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcggta cctttaagac 3781 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg 3841 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct 3901 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc 3961 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg 4021 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt 4081 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga 4141 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4201 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4261 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc 4321 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 4381 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 4441 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt 4501 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 4561 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 4621 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt 4681 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 4741 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 4801 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 4861 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 4921 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 4981 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 5041 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt 5101 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 5161 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 5221 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 5281 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 5341 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 5401 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 5461 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5521 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5581 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5641 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5701 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5761 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5821 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5881 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5941 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 6001 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 6061 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 6121 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 6181 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 6241 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 6301 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 6361 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 6421 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6481 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6541 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6601 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6661 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6721 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6781 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6841 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6901 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6961 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 7021 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 7081 aatg //