LOCUS V020700 8174 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020700 VERSION V020700 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..2854 /label="PIK3R1(NM_181523)" /note="PIK3R1(NM_181523)" /gene="PIK3R1" CDS 2861..2926 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2950..3538 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3604..3652 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3854..4282 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4296..4625 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4692..5486 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5662..5783 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5832..5848) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5856..5872 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(5880..5910) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5925..5946 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6234..6822) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6993..7853) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(7854..7958) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatgagtgc tgaggggtac cagtacagag cgctgtatga 721 ttataaaaag gaaagagaag aagatattga cttgcacttg ggtgacatat tgactgtgaa 781 taaagggtcc ttagtagctc ttggattcag tgatggacag gaagccaggc ctgaagaaat 841 tggctggtta aatggctata atgaaaccac aggggaaagg ggggactttc cgggaactta 901 cgtagaatat attggaagga aaaaaatctc gcctcccaca ccaaagcccc ggccacctcg 961 gcctcttcct gttgcaccag gttcttcgaa aactgaagca gatgttgaac aacaagcttt 1021 gactctcccg gatcttgcag agcagtttgc ccctcctgac attgccccgc ctcttcttat 1081 caagctcgtg gaagccattg aaaagaaagg tctggaatgt tcaactctat acagaacaca 1141 gagctccagc aacctggcag aattacgaca gcttcttgat tgtgatacac cctccgtgga 1201 cttggaaatg atcgatgtgc acgttttggc tgacgctttc aaacgctatc tcctggactt 1261 accaaatcct gtcattccag cagccgttta cagtgaaatg atttctttag ctccagaagt 1321 acaaagctcc gaagaatata ttcagctatt gaagaagctt attaggtcgc ctagcatacc 1381 tcatcagtat tggcttacgc ttcagtattt gttaaaacat ttcttcaagc tctctcaaac 1441 ctccagcaaa aatctgttga atgcaagagt actctctgaa attttcagcc ctatgctttt 1501 cagattctca gcagccagct ctgataatac tgaaaacctc ataaaagtta tagaaatttt 1561 aatctcaact gaatggaatg aacgacagcc tgcaccagca ctgcctccta aaccaccaaa 1621 acctactact gtagccaaca acggtatgaa taacaatatg tccttacaag atgctgaatg 1681 gtactgggga gatatctcga gggaagaagt gaatgaaaaa cttcgagata cagcagacgg 1741 gacctttttg gtacgagatg cgtctactaa aatgcatggt gattatactc ttacactaag 1801 gaaaggggga aataacaaat taatcaaaat atttcatcga gatgggaaat atggcttctc 1861 tgacccatta accttcagtt ctgtggttga attaataaac cactaccgga atgaatctct 1921 agctcagtat aatcccaaat tggatgtgaa attactttat ccagtatcca aataccaaca 1981 ggatcaagtt gtcaaagaag ataatattga agctgtaggg aaaaaattac atgaatataa 2041 cactcagttt caagaaaaaa gtcgagaata tgatagatta tatgaagaat atacccgcac 2101 atcccaggaa atccaaatga aaaggacagc tattgaagca tttaatgaaa ccataaaaat 2161 atttgaagaa cagtgccaga cccaagagcg gtacagcaaa gaatacatag aaaagtttaa 2221 acgtgaaggc aatgagaaag aaatacaaag gattatgcat aattatgata agttgaagtc 2281 tcgaatcagt gaaattattg acagtagaag aagattggaa gaagacttga agaagcaggc 2341 agctgagtat cgagaaattg acaaacgtat gaacagcatt aaaccagacc ttatccagct 2401 gagaaagacg agagaccaat acttgatgtg gttgactcaa aaaggtgttc ggcaaaagaa 2461 gttgaacgag tggttgggca atgaaaacac tgaagaccaa tattcactgg tggaagatga 2521 tgaagatttg ccccatcatg atgagaagac atggaatgtt ggaagcagca accgaaacaa 2581 agctgaaaac ctgttgcgag ggaagcgaga tggcactttt cttgtccggg agagcagtaa 2641 acagggctgc tatgcctgct ctgtagtggt ggacggcgaa gtaaagcatt gtgtcataaa 2701 caaaacagca actggctatg gctttgccga gccctataac ttgtacagct ctctgaaaga 2761 actggtgcta cattaccaac acacctccct tgtgcagcac aacgactccc tcaatgtcac 2821 actagcctac ccagtatatg cacagcagag gcgactcgag gactacaaag accatgacgg 2881 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga 2941 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 3001 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 3061 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 3121 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 3181 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 3241 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 3301 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 3361 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 3421 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 3481 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 3541 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 3601 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 3661 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 3721 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 3781 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 3841 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3901 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 4201 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 4261 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 4561 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 4621 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 4681 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 4741 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 4801 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 4861 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4921 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4981 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 5041 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 5101 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 5161 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 5221 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 5281 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 5341 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 5401 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 5461 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 5521 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 5581 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 5641 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 5701 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 5761 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 5821 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5881 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5941 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 6001 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 6061 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6421 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 6541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 6661 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 6721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 7261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 7561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 7681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 7741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct 8041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 8101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 8161 agaatctgct tagg //