LOCUS Exported 9017 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.1 myc TSC1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9017) AUTHORS Cai SL, Tee AR, Short JD, Bergeron JM, Kim J, Shen J, Guo R, Johnson CL, Kiguchi K, Walker CL TITLE Activity of TSC2 is inhibited by AKT-mediated phosphorylation and membrane partitioning. JOURNAL J Cell Biol. 2006 Apr 24. 173(2):279-89. PUBMED 16636147 REFERENCE 2 (bases 1 to 9017) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9017 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 4502..4531 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 4547..4564 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4608..4625) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4614..4838 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4884..5312 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4971..4990) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5181..5202 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5321..5341) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5326..5655 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5506..5641 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5568..5587 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5722..6516 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5776..5795) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6386..6405 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6690..6811 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6727..6746) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6781..6800 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6860..6876) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6860..6876) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6873..6895) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6884..6900 /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)." promoter complement(6908..6938) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6953..6974 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7091..7108) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7262..7850) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7342..7361) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8021..8881) /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 8644..8663 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8882..8986) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc 961 accacactgg actagtggat ccgagctcgg taccatggcc caacaagcaa atgtcgggga 1021 gcttcttgcc atgctggact cccccatgct gggtgtgcgg gacgacgtga cagctgtctt 1081 taaagagaac ctcaattctg accgtggccc tatgcttgta aacaccttgg tggattatta 1141 cctggaaacc agctctcagc cggcattgca catcctgacc accttgcaag agccacatga 1201 caagcacctc ttggacagga ttaacgaata tgtgggcaaa gccgccactc gtttatccat 1261 cctctcgtta ctgggtcatg tcataagact gcagccatct tggaagcata agctctctca 1321 agcacctctt ttgccttctt tactaaaatg tctcaagatg gacactgacg tcgttgtcct 1381 cacaacaggc gtcttggtgt tgataaccat gctaccaatg attccacagt ctgggaaaca 1441 gcatcttctt gatttctttg acatttttgg ccgtctgtca tcatggtgcc tgaagaaacc 1501 aggccacgtg gcggaagtct atctcgtcca tctccatgcc agtgtgtacg cactctttca 1561 tcgcctttat ggaatgtacc cttgcaactt cgtctccttt ttgcgttctc attacagtat 1621 gaaagaaaac ctggagactt ttgaagaagt ggtcaagcca atgatggagc atgtgcgaat 1681 tcatccggaa ttagtgactg gatccaagga ccatgaactg gaccctcgaa ggtggaagag 1741 attagaaact catgatgttg tgatcgagtg tgccaaaatc tctctggatc ccacagaagc 1801 ctcatatgaa gatggctatt ctgtgtctca ccaaatctca gcccgctttc ctcatcgttc 1861 agccgatgtc accaccagcc cttatgctga cacacagaat agctatgggt gtgctacttc 1921 taccccttac tccacgtctc ggctgatgtt gttaaatatg ccagggcagc tacctcagac 1981 tctgagttcc ccatcgacac ggctgataac tgaaccacca caagctactc tttggagccc 2041 atctatggtt tgtggtatga ccactcctcc aacttctcct ggaaatgtcc cacctgatct 2101 gtcacaccct tacagtaaag tctttggtac aactgcaggt ggaaaaggaa ctcctctggg 2161 aaccccagca acctctcctc ctccagcccc actctgtcat tcggatgact acgtgcacat 2221 ttcactcccc caggccacag tcacaccccc caggaaggaa gagagaatgg attctgcaag 2281 accatgtcta cacagacaac accatcttct gaatgacaga ggatcagaag agccacctgg 2341 cagcaaaggt tctgtcactc taagtgatct tccagggttt ttaggtgatc tggcctctga 2401 agaagatagt attgaaaaag ataaagaaga agctgcaata tctagagaac tttctgagat 2461 caccacagca gaggcagagc ctgtggttcc tcgaggaggc tttgactctc ccttttaccg 2521 agacagtctc ccaggttctc agcggaagac ccactcggca gcctccagtt ctcagggcgc 2581 cagcgtgaac cctgagcctt tacactcctc cctggacaag cttgggcctg acacaccaaa 2641 gcaagccttt actcccatag acctgccctg cggcagtgct gatgaaagcc ctgcgggaga 2701 cagggaatgc cagacttctt tggagaccag tatcttcact cccagtcctt gtaaaattcc 2761 acctccgacg agagtgggct ttggaagcgg gcagcctccc ccgtatgatc atctttttga 2821 ggtggcattg ccaaagacag cccatcattt tgtcatcagg aagactgagg agctgttaaa 2881 gaaagcaaaa ggaaacacag aggaagatgg tgtgccctct acctccccaa tggaagtgct 2941 ggacagactg atacagcagg gagcagacgc gcacagcaag gagctgaaca agttgccttt 3001 acccagcaag tctgtcgact ggacccactt tggaggctct cctccttcag atgagatccg 3061 caccctccga gaccagttgc ttttactgca caaccagtta ctctatgagc gttttaagag 3121 gcagcagcat gccctccgga acaggcggct cctccgcaag gtgatcaaag cagcagctct 3181 ggaggaacat aatgctgcca tgaaagatca gttgaagtta caagagaagg acatccagat 3241 gtggaaggtt agtctgcaga aagaacaagc tagatacaat cagctccagg agcagcgtga 3301 cactatggta accaagctcc acagccagat cagacagctg cagcatgacc gagaggaatt 3361 ctacaaccag agccaggaat tacagacgaa gctggaggac tgcaggaaca tgattgcgga 3421 gctgcggata gaactgaaga aggccaacaa caaggtgtgt cacactgagc tgctgctcag 3481 tcaggtttcc caaaagctct caaacagtga gtcggtccag cagcagatgg agttcttgaa 3541 caggcagctg ttggttcttg gggaggtcaa cgagctctat ttggaacaac tgcagaacaa 3601 gcactcagat accacaaagg aagtagaaat gatgaaagcc gcctatcgga aagagctaga 3661 aaaaaacaga agccatgttc tccagcagac tcagaggctt gatacctccc aaaaacggat 3721 tttggaactg gaatctcacc tggccaagaa agaccacctt cttttggaac agaagaaata 3781 tctagaggat gtcaaactcc aggcaagagg acagctgcag gccgcagaga gcaggtatga 3841 ggctcagaaa aggataaccc aggtgtttga attggagatc ttagatttat atggcaggtt 3901 ggagaaagat ggcctcctga aaaaacttga agaagaaaaa gcagaagcag ctgaagcagc 3961 agaagaaagg cttgactgtt gtaatgacgg gtgctcagat tccatggtag ggcacaatga 4021 agaggcatct ggccacaacg gtgagaccaa gacccccagg cccagcagcg cccggggcag 4081 tagtggaagc agaggtggtg gaggcagcag cagcagcagc gagctttcta ccccagagaa 4141 acccccacac cagagggcag gcccattcag cagtcggtgg gagacgacta tgggagaagc 4201 gtctgccagc atccccacca ctgtgggctc acttcccagt tcaaaaagct tcctgggtat 4261 gaaggctcga gagttatttc gtaataagag cgagagccag tgtgatgagg acggcatgac 4321 cagtagcctt tctgagagcc taaagacaga actgggcaaa gacttgggtg tggaagccaa 4381 gattcccctg aacctagatg gccctcaccc gtctcccccg accccggaca gtgttggaca 4441 gctacatatc atggactaca atgagactca tcatgaacac agcggtacca agcttgggcc 4501 cgaacaaaaa ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca 4561 tcattgagtt taaacggtct ccagcttaag tttaaaccgc tgatcagcct cgactgtgcc 4621 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 4681 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 4741 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 4801 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 4861 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 4921 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 4981 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 5041 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 5101 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 5161 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 5221 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 5281 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 5341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 5401 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 5461 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 5521 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 5581 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 5641 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 5701 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 5761 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 5821 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 5881 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 5941 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 6001 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 6061 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 6121 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 6181 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 6241 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 6301 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 6361 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 6421 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 6481 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 6541 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 6601 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 6661 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 6721 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 6781 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 6841 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6901 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6961 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7021 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7081 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7141 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7201 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7681 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 7741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7861 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7921 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7981 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8041 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8101 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8161 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8221 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8281 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8341 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8401 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8461 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8521 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8581 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8641 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8701 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8761 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8821 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8881 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8941 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9001 aagtgccacc tgacgtc //