LOCUS Exported 7568 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS PIK3CA-WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7568) AUTHORS Samuels Y, Wang Z, Bardelli A, Silliman N, Ptak J, Szabo S, Yan H, Gazdar A, Powell SM, Riggins GJ, Willson JK, Markowitz S, Kinzler KW, Vogelstein B, Velculescu VE TITLE High frequency of mutations of the PIK3CA gene in human cancers. JOURNAL Science. 2004 Apr 23. 304(5670):554. PUBMED 15016963 REFERENCE 2 (bases 1 to 7568) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7568 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 526..546 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 621..639 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" regulatory 674..683 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 683..706 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(4018..4037) /label=T7 /note="T7 promoter, forward primer" promoter complement(4019..4037) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 4311..4432 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4348..4367) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4402..4421 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4439..4894) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4576..4597) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4788..4807 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4921..5025 /gene="bla" /label=AmpR promoter promoter 5027..5384 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5050..5070) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5235..5370 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5297..5316 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5419..6213 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5473..5492) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6083..6102 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6401..6420) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6445..6492 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6821..7409 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7310..7329 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 601 agcgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac 661 cgcggtggcg gccgccacca tggattacaa ggatgacgac gataaggccc gggcggatcc 721 cccgggctgc aggaattcga tagttcgaac catgcctcca agaccatcat caggtgaact 781 gtggggcatc cacttgatgc ccccaagaat cctagtagaa tgtttactac caaatggaat 841 gatagtgact ttagaatgcc tccgcgaggc tacattaata accataaagc atgaactatt 901 taaagaagca agaaaatacc ccctccatca acttcttcaa gatgaatctt cttacatttt 961 cgtaagtgtt actcaagaag cagaaaggga agaatttttt gatgaaacaa gacgactttg 1021 tgaccttcgg ctttttcaac cctttttaaa agtaattgaa ccagtaggca accgtgaaga 1081 aaagatcctc gatcgagaaa ttggttttgc tatcggcatg ccagtgtgtg aatttgatat 1141 ggttaaagat ccagaagtac aggacttccg aagaaatatt ctgaacgttt gtaaagaagc 1201 tgtggatctt agggacctca attcacctca tagtagagca atgtatgtct atcctccaaa 1261 tgtagaatct tcaccagaat cgccaaagca catatacaat aaattagata aagggcaaat 1321 aatagtggtg atctgggtaa tagtttctcc aaataatgac aagcagaagt atactctgaa 1381 aatcaaccat gactgtgtac cagaacaagt aattgctgaa gcaatcagga aaaaaactcg 1441 aagtatgttg ctatcctctg aacaactaaa actctgtgtt ttagaatatc agggcaagta 1501 tattttaaaa gtgtgtggat gtgatgaata cttcctagaa aaatatcctc tgagtcagta 1561 taagtatata agaagctgta taatgctagg gaggatgccc aatttgatgt tgatggctaa 1621 agaaagcctt tattctcaac tgccgatgga ctgtttcaca atgccatctt attccagacg 1681 catttccaca gctacaccat atatgaatgg agaagcatct acaaaatccc tttgggttat 1741 aaatagtgca ctcagaataa aaattctttg tgcaacctac gtgaatgtaa atattcgaga 1801 cattgataag atctatgttc gaacaggtat ctaccatgga ggagaaccct tatgtgacaa 1861 tgtgaacact caaagagtac cttgttccaa tcccaggtgg aatgaatggc tgaattatga 1921 tatatacatt cctgatcttc ctcgtgctgt tcgactttgc ccttccattt gctctgttaa 1981 aggccgaaag ggtgctaaag aggaacactg tccattggca tggggaaata taaacttgtt 2041 tgattacaca gacactctag tatctggaaa aatggctttg aatctttggc cagtacctca 2101 tggattagaa gatttgctga accctattgg tgttactgga tcaaatccaa ataaagaaac 2161 tccatgctta gagttggagt ttgactggtt cagcagtgtg gtaaagttcc cagatatgtc 2221 agtgattgaa gagcatgcca attggtctgt atcccgagaa gcaggattta gctattccca 2281 cgcaggactg agtaacagac tagctagaga caatgaatta agggaaaatg acaaagaaca 2341 gctcaaagca atttctacac gagatcctct ctctgaaatc actgagcagg agaaagattt 2401 tctatggagt cacagacact attgtgtaac tatccccgaa attctaccca aattgcttct 2461 gtctgttaaa tggaattcta gagatgaagt agcccagatg tattgcttgg taaaagattg 2521 gcctccaatc aaacctgaac aggctatgga acttctggac tgtaattacc cagatcctat 2581 ggttcgaggt tttgctgttc ggtgcttgga aaaatattta acagatgaca aactttctca 2641 gtatttaatt cagctagtac aggtcctaaa atatgaacaa tatttggata acttgcttgt 2701 gagattttta ctgaagaaag cattgactaa tcaaaggatt gggcactttt tcttttggca 2761 tttaaaatct gagatgcaca ataaaacagt tagccagagg tttggcctgc ttttggagtc 2821 ctattgtcgt gcatgtggga tgtatttgaa gcacctgaat aggcaagtcg aggcaatgga 2881 aaagctcatt aacttaactg acattctcaa acaggagaag aaggatgaaa cacaaaaggt 2941 acagatgaag tttttagttg agcaaatgag gcgaccagat ttcatggatg ctctacaggg 3001 ctttctgtct cctctaaacc ctgctcatca actaggaaac ctcaggcttg aagagtgtcg 3061 aattatgtcc tctgcaaaaa ggccactgtg gttgaattgg gagaacccag acatcatgtc 3121 agagttactg tttcagaaca atgagatcat ctttaaaaat ggggatgatt tacggcaaga 3181 tatgctaaca cttcaaatta ttcgtattat ggaaaatatc tggcaaaatc aaggtcttga 3241 tcttcgaatg ttaccttatg gttgtctgtc aatcggtgac tgtgtgggac ttattgaggt 3301 ggtgcgaaat tctcacacta ttatgcaaat tcagtgcaaa ggcggcttga aaggtgcact 3361 gcagttcaac agccacacac tacatcagtg gctcaaagac aagaacaaag gagaaatata 3421 tgatgcagcc attgacctgt ttacacgttc atgtgctgga tactgtgtag ctaccttcat 3481 tttgggaatt ggagatcgtc acaatagtaa catcatggtg aaagacgatg gacaactgtt 3541 tcatatagat tttggacact ttttggatca caagaagaaa aaatttggtt ataaacgaga 3601 acgtgtgcca tttgttttga cacaggattt cttaatagtg attagtaaag gagcccaaga 3661 atgcacaaag acaagagaat ttgagaggtt tcaggagatg tgttacaagg cttatctagc 3721 tattcgacag catgccaatc tcttcataaa tcttttctca atgatgcttg gctctggaat 3781 gccagaacta caatcttttg atgacattgc atacattcga aagaccctag ccttagataa 3841 aactgagcaa gaggctttgg agtatttcat gaaacaaatg aatgatgcac atcatggtgg 3901 ctggacaaca aaaatggatt ggatcttcca cacaattaaa cagcatgcat tgaactagct 3961 cgaggggggg cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc 4021 tatagtgagt cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat 4081 ggcgaatgga gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt 4141 gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt 4201 tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 4261 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 4321 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4381 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa 4441 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt 4501 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata 4561 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac 4621 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa 4681 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc 4741 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg 4801 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca 4861 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg 4921 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 4981 caataaccct gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga 5041 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 5101 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 5161 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 5221 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 5281 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc 5341 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca 5401 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 5461 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 5521 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 5581 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc 5641 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 5701 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 5761 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 5821 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 5881 caggatgatc tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc 5941 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 6001 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 6061 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc 6121 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 6181 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 6241 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 6301 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 6361 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa 6421 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg 6481 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac 6541 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa 6601 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct 6661 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 6721 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 6781 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 6841 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 6901 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 6961 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 7021 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 7081 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 7141 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 7201 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 7261 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 7321 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 7381 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 7441 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 7501 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt 7561 catagccc //