LOCUS Exported 8227 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-myc3-CUL4B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8227) 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 8227) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8227 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter primer_bind complement(1981..2000) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2172..2551 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2552..2755 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2706..2726 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2800..2819 /label=T7 /note="T7 promoter, forward primer" promoter 2800..2818 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2834..2863 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2870..2899 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2906..2935 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" promoter complement(5717..5735) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5718..5735) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5755..5772) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5761..5985 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 6031..6459 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6118..6137) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6328..6349 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6468..6488) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6473..6802 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6653..6788 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6715..6734 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6869..7663 /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(6923..6942) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7533..7552 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 7837..7958 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7874..7893) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7928..7947 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8007..8023) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8007..8023) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8020..8042) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8031..8047 /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(8055..8085) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8100..8121 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag 1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga 2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa 2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag 2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt 2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg 2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg 2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc 2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg 2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga 2821 gacccaagct atggagcaaa agctcatctc agaggaagat ctggaattag agcaaaagct 2881 catctcagag gaagatctgg aattagagca aaagctcatc tcagaggaag atctggaatt 2941 caggatccct atgtcacagt catctggatc aggagatggg aatgatgatg aggctactac 3001 ctctaaagac ggtggttttt cttcccccag tccctcagct gctgctgctg ctcaggaggt 3061 cagatctgcc actgatggta ataccagcac cactccgccc acctctgcca agaagagaaa 3121 gttaaacagc agcagcagta gcagcagtaa cagtagtaac gagagagaag actttgattc 3181 cacctcttcc tcctcttcca ctcctccttt acaacccagg gattcggcat ccccttcaac 3241 ctcgtccttc tgcctggggg tttcagtggc tgcttccagc cacgtaccga tacagaagaa 3301 gctgcgtttt gaagacaccc tggagtttgt agggtttgat gcgaagatgg ctgaggaatc 3361 ctcctcctcc tcctcctcat cttcaccaac tgctgcaaca tctcagcagc agcaacttaa 3421 aaataagagt atattaatct cttctgtggc ttcggtgcat catgcaaacg gcctagccaa 3481 atcttctacc accgtctcta gctttgctaa cagcaaacct ggctctgcta agaagttagt 3541 gatcaagaac tttaaagata agcctaaatt accagaaaac tacacagatg aaacctggca 3601 aaaactgaaa gaagcagtgg aagctattca gaatagtact tcaattaagt acaatttaga 3661 agaactctac caggctgtag aaaatctctg ttcttacaag atttctgcaa acttgtacaa 3721 acagctgaga cagatctgcg aagatcacat caaagcacag attcatcaat tcagagagga 3781 ttcattggat agcgttcttt ttttaaagaa gattgataga tgctggcaaa accattgcag 3841 acaaatgatc atgatcagga gcattttttt gtttctggat agaacttacg ttcttcagaa 3901 ttcaatgcta ccctccattt gggacatggg actggagtta tttagggctc atattataag 3961 tgatcagaaa gtgcagaata agacaattga tggcattctt ctcttgattg agagggaaag 4021 gaatggtgaa gcaattgata gaagtttact tcgaagcctt ttaagcatgc tgtctgattt 4081 gcaaatttat caagattctt ttgaacaacg atttttggaa gaaactaacc ggctctatgc 4141 agctgaaggc caaaaattaa tgcaagaaag agaggttcct gaatatctac atcatgttaa 4201 caaacgtcta gaagaagaag cagacagact tattacttac ttagatcaga ccacccagaa 4261 gtcattaatt gctactgtag aaaaacaact tctaggtgaa cacttaacag caattcttca 4321 gaaaggttta aataacctcc ttgatgaaaa ccgaattcaa gatttgtctc ttctgtatca 4381 gctcttcagt agagttcgag gtggagttca ggttcttttg cagcagtgga tcgaatatat 4441 caaggcattt ggcagcacta ttgtaattaa tcctgaaaaa gataaaacca tggttcaaga 4501 attgctggat tttaaagata aggttgacca tataattgat atctgctttc tgaagaatga 4561 gaaatttatc aatgccatga aagaagcatt tgaaacgttc attaacaaaa gaccaaataa 4621 accagctgaa cttatagcta agtatgtaga ttcaaaactt cgtgcaggca acaaagaagc 4681 tacagatgaa gaacttgaga aaatgttgga taaaattatg atcatattta gatttatcta 4741 tggcaaggat gtttttgagg ccttctataa gaaagattta gccaagcgcc tgttagtcgg 4801 aaagagtgca tctgtagatg ctgaaaaatc aatgctgtcc aaacttaaac atgaatgcgg 4861 agctgctttc accagcaaac ttgaaggaat gtttaaagac atggaacttt ctaaagacat 4921 catgattcag ttcaaacagt atatgcagaa tcagaatgtt ccgggaaata ttgagttaac 4981 tgtgaatatc ctgacaatgg gctattggcc gacatatgtg cctatggaag ttcatttacc 5041 accagagatg gtaaaacttc aggagatttt caagacattt tacctaggca aacatagtgg 5101 caggaaactt cagtggcagt caaccatagg acactgtgtg ttaaaagcag aatttaaaga 5161 gggtaaaaag gaactccagg tctctctttt tcaaacactg gtgctgctaa tgtttaatga 5221 gggagaggag ttcagtttag aagagatcaa gcaggcaact ggaatagagg atggagagtt 5281 aaggagaaca ctgcagtcat tagcctgtgg caaagctaga gttctggcga aaaatccaaa 5341 gggcaaagac attgaagatg gtgacaagtt catttgtaat gatgatttca aacataaact 5401 tttcaggata aagatcaatc aaatccagat gaaagaaacg gttgaagaac aagcaagcac 5461 tacagaaaga gtatttcaag acagacagta tcaaattgat gctgcaattg ttcgaattat 5521 gaagatgaga aagacactta gccacaatct ccttgtttca gaagtgtaca accagttgaa 5581 atttccagta aagcctgctg atcttaagaa gagaatagaa tctttaattg accgggacta 5641 catggaaaga gataaagaaa atccaaacca gtacaactat attgcatagc tcgagcatgc 5701 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga 5761 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 5821 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 5881 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 5941 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 6001 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg 6061 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 6121 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 6181 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 6241 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 6301 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 6361 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 6421 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca 6481 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 6541 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 6601 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 6661 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6721 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 6781 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga 6841 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 6901 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 6961 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 7021 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 7081 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 7141 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 7201 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 7261 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 7321 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 7381 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 7441 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 7501 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 7561 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 7621 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 7681 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 7741 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 7801 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 7861 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 7921 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 7981 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 8041 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 8101 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 8161 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 8221 gggcgct //