LOCUS Exported 7122 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCDNA3-myc3-FBW5 delta F SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7122) AUTHORS Hu J, Zacharek S, He YJ, Lee H, Shumway S, Duronio RJ, Xiong Y TITLE WD40 protein FBW5 promotes ubiquitination of tumor suppressor TSC2 by DDB1-CUL4-ROC1 ligase. JOURNAL Genes Dev. 2008 Apr 1. 22(7):866-71. PUBMED 18381890 REFERENCE 2 (bases 1 to 7122) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7122 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 899..928 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 935..964 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 971..1000 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2600..2665 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" promoter complement(2677..2695) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2678..2695) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2715..2732) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2721..2945 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2991..3419 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3078..3097) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3288..3309 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3428..3448) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3433..3762 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3613..3748 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3675..3694 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3829..4623 /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(3883..3902) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4493..4512 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4797..4918 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4834..4853) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4888..4907 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4967..4983) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4967..4983) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4980..5002) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4991..5007 /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(5015..5045) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5060..5081 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5198..5215) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5369..5957) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5449..5468) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6128..6988) /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 6751..6770 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6989..7093) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctatgga 901 gcaaaagctc atctcagagg aagatctgga attagagcaa aagctcatct cagaggaaga 961 tctggaatta gagcaaaagc tcatctcaga ggaagatctg gaattcatgt ggagggagca 1021 gttctaccgc tactaccagg tggcccgcga cgtgccccga cacccagcgg ccatgtcctg 1081 gtacgaggag ttccagcggc tgtatgacac ggtgccctgc gtggaggtgc agacgctgcg 1141 ggaacacaca gaccaggtcc tgcacctcag cttctcccat tccgggtacc agttcgcgtc 1201 ctgctccaag gactgcactg tgaagatctg gagcaacgac ctgaccatct cgctgctgca 1261 cagcgcggac atgcggccct acaactggag ctacacccag ttctcccagt tcaacaagga 1321 cgactcgcta ctgctggcct cgggggtgtt cctggggccg cacaactcct catccggcga 1381 gattgctgtc atcagcctag actccttcgc gctgctgtcc cgcgtgcgga acaagcccta 1441 tgacgtgttt ggctgttggc tcaccgagac cagcctcatc tcggggaacc tgcaccgcat 1501 cggagatatc acctcctgct cggtgctgtg gctcaacaat gccttccagg atgtggagtc 1561 agagaacgtc aacgtggtga agcggctgtt caagatccag aacctcaatg ccagcaccgt 1621 ccgcacggtg atggtggccg actgcagccg cttcgacagc cctgacctgc tgctggaagc 1681 cggtgacccg gccacgtccc cctgccgcat ctttgacctg ggcagcgaca acgaggaggt 1741 ggtggctggc ccggcccccg cccacgccaa ggagggcttg cggcactttc tggatcgcgt 1801 gctggagggg cgggcgcagc cacagctgtc ggagcgcatg ctagagacca aggtggccga 1861 gctgctggcc cagggccaca ccaagccacc cgagcgcagt gccacaggcg ccaagagcaa 1921 gtacctcatc ttcaccactg gctgcctcac ctactcccca caccagatcg gcatcaagca 1981 gatcctgcca caccagatga ccacggcagg gcccgtgctg ggtgagggcc ggggctccga 2041 tgccttcttc gacgcgctgg accacgtcat agacatacac ggacacatca tcggcatggg 2101 cctgtcgccc gacaacaggt acctgtacgt gaacagccgc gcctggccca acggtgcggt 2161 ggtggccgac cccatgcagc cgccaccaat cgcggaggag attgacctgc tggtgttcga 2221 cctcaagacc atgcgggagg tgaggcgggc tctgcgtgcg caccgcgcct acacgcccaa 2281 cgacgagtgc ttcttcatct tcctggacgt cagcagggac ttcgtggcca gcggggcgga 2341 ggaccggcac ggctacatct gggaccgcca ctacaacatc tgtctggcca ggctgcggca 2401 cgaggatgtg gtcaactcag tggtcttcag tccccaggag caggagctgc tgctcacggc 2461 cagcgacgac gccaccatca aagcctggcg ctccccacgc accatgcgcg tcctccaggc 2521 acctcgccca cggcctcgca ccttcttctc ctggcttgcc agccagaggc gctcgagcat 2581 gcatctagag ttaaccatgg actacaaaga ccatgacggt gattataaag atcatgatat 2641 cgattacaag gatgacgatg acaagtaggg ccctattcta tagtgtcacc taaatgctag 2701 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc 2761 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 2821 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 2881 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 2941 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg 3001 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 3061 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 3121 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 3181 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 3241 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 3301 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 3361 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 3421 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 3481 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 3601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 3661 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 3721 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 3781 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 3841 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 3901 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 3961 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 4021 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 4081 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 4141 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 4201 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 4261 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 4321 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 4381 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 4441 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 4501 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 4561 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 4621 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 4681 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 4741 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 4801 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 4861 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 4921 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 4981 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 5041 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 5101 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 5161 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 5221 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 5281 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 5341 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 5401 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 5461 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 5521 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 5581 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 5641 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 5701 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 5761 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 5821 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 5881 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 5941 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 6001 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 6061 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 6121 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 6181 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 6241 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 6301 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6361 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6421 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6481 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6541 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6601 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6661 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6721 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6781 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6841 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6901 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6961 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 7021 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 7081 taggggttcc gcgcacattt ccccgaaaag tgccacctga cg //