LOCUS Exported 8242 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-myc3-CUL4A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8242) AUTHORS Liu J, Furukawa M, Matsumoto T, Xiong Y TITLE NEDD8 modification of CUL1 dissociates p120(CAND1), an inhibitor of CUL1-SKP1 binding and SCF ligases. JOURNAL Mol Cell. 2002 Dec . 10(6):1511-8. PUBMED 12504025 REFERENCE 2 (bases 1 to 8242) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8242 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 137..516 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 517..720 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 671..691 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 765..784 /label=T7 /note="T7 promoter, forward primer" promoter 765..783 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 799..828 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 835..864 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 871..900 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" promoter complement(3697..3715) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3698..3715) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3735..3752) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3741..3965 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4011..4439 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4098..4117) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4308..4329 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4448..4468) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4453..4782 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4633..4768 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4695..4714 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4849..5643 /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(4903..4922) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5513..5532 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5817..5938 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5854..5873) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5908..5927 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5987..6003) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5987..6003) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6000..6022) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6011..6027 /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(6035..6065) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6080..6101 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6218..6235) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6389..6977) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6469..6488) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7148..8008) /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 7771..7790 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8009..8113) /gene="bla" /label=AmpR promoter primer_bind complement(8188..8207) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc 61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg 121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg 181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg 241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata 301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc 361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac 421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc 541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc 601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc 661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct 721 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat 781 agggagaccc aagctatgga gcaaaagctc atctcagagg aagatctgga attagagcaa 841 aagctcatct cagaggaaga tctggaatta gagcaaaagc tcatctcaga ggaagatctg 901 gaattcagga tcccccgggc tgcaggaatt cggcacgagc cagccatggc ggacgaggcc 961 ccgcggaagg gcagcttctc ggcgctcgtg ggccgcacca acggcctcac caagcccgcg 1021 gccctggccg ccgcgcccgc caagccgggg ggcgcgggcg gctccaagaa gctggtcatc 1081 aagaacttcc gagacagacc tcggctgccc gacaactaca cgcaggacac gtggcggaag 1141 ctgcacgagg cggtgcgggc cgtgcagagc agcacctcca tcaggtacaa cctcgaggag 1201 ctctaccagg ctgtggaaaa tctctgttct cacaaagtct ccccaatgct ctacaagcaa 1261 ctgcgtcagg cctgtgaaga ccacgtccag gcacagatcc ttccgtttag agaagactca 1321 ctagatagtg ttttattttt aaagaagatt aacacgtgct ggcaggacca ctgcagacaa 1381 atgatcatga tcagaagcat cttcctgttc ttggaccgca cctatgtgct gcagaactcc 1441 acgctgccct ccatctggga tatgggatta gaactgttta gaacccatat tattagtgat 1501 aaaatggttc agagtaaaac cattgatgga atcctactgc tgatcgagcg cgagaggagc 1561 ggcgaggccg tggaccggag cctgttgcgg agcctcctgg gcatgctgtc tgacctgcag 1621 gtgtataaag attcatttga actgaaattt ttggaagaga ctaattgctt atatgctgcc 1681 gaaggccaaa ggttaatgca ggaaagagag gtcccagaat atcttaacca tgtaagtaaa 1741 cgcttagagg aagagggaga cagagtaatc acttacttgg accacagcac acagaaacca 1801 ctgattgctt gtgtggagaa acagctatta ggagaacatt taacagcaat tctgcagaaa 1861 gggctcgacc acttactgga tgagaacaga gtgccggacc tcgcacagat gtaccagctg 1921 ttcagccggg tgaggggcgg gcagcaggcg ctgctgcagc actggagcga gtacatcaag 1981 acttttggaa cagcgatcgt aatcaatcct gagaaagata aagacatggt ccaagacctg 2041 ttggacttca aggacaaggt ggaccacgtg atcgaggtct gcttccagaa gaatgagcgg 2101 ttcgtcaacc tgatgaagga gtcctttgag acgttcatca acaagagacc caacaagcct 2161 gcagaactga tcgcaaagca tgtggattca aagttaagag caggcaacaa agaagccaca 2221 gacgaggagc tggagcggac gttggacaag atcatgatcc tgttcaggtt tatccacggt 2281 aaagatgtct ttgaagcatt ttataaaaaa gatttggcaa aaagactcct tgttgggaaa 2341 agtgcctcag tcgatgctga aaagtctatg ttgtcaaagc tcaagcatga gtgcggtgca 2401 gccttcacca gcaagctgga aggcatgttc aaggacatgg agctttcgaa ggacatcatg 2461 gttcatttca agcagcatat gcagaatcag agtgactcag gccctataga cctcacagtg 2521 aacatactca caatgggcta ctggccaaca tacacgccca tggaagtgca cttaacccca 2581 gaaatgatta aacttcagga agtatttaag gcattttatc ttggaaagca cagtggtcga 2641 aaacttcagt ggcaaactac tttgggacat gctgttttaa aagcggagtt taaagaaggg 2701 aagaaggaat tccaggtgtc cctcttccag acactggtgc tcctcatgtt caacgaggga 2761 gatggcttca gctttgagga gataaaaatg gccacgggga tagaggatag tgaattgcgc 2821 agaacgctgc agtccctggc ctgtggcaaa gcacgtgtgc tgattaaaag tcccagagga 2881 aaggaagtgg aagatggaga caagttcatt tttaatggag agttcaagca caagttgttt 2941 agaataaaga tcaatcaaat tcagatgaag gaaactgttg aggaacaggt tagcaccact 3001 gagagagtgt ttcaggatag acaatatcag attgatgctg ctatcgtcag aataatgaag 3061 atgagaaaga ctcttggtca taatcttcta gtttctgaat tatataatca gctgaaattt 3121 ccagtaaagc ctggagattt gaaaaagaga attgaatctc tgatagacag agactatatg 3181 gagagagaca aagacaatcc gaatcagtac cactacgtgg cctgacgcat ctgcagacgg 3241 ttccccttca tgaaacacta gaatgtaccc tcagagcagg aagcacacct gtgccatttc 3301 tgggactctg attgatccag ctgtggacat tggaaggcga aggaagggag gtggctcctg 3361 ggtcatcttt cacaaggctc aagacttcaa cctgcagatg tatctttttc cctccagttt 3421 ttcctctagt tcttttaggc atttaaattg tttctgttac tctgtgcaaa ataactttga 3481 gattggacaa gaagatgtta ctaaagagaa gttcctttaa aaggtcttgt tcttgtgtca 3541 aaaagctgca agtttggttt gttctcgtgt gtgatcatga gtgcacaatg aagaagaccc 3601 tagatgctgc attttttagc tctgaagatt ccttaggtat ccctgaagac agctcgctca 3661 gatgatcagc atttagagtg aaaacaaggg ccctattcta tagtgtcacc taaatgctag 3721 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc 3781 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 3841 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 3901 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 3961 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg 4021 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 4081 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 4141 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 4201 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 4261 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 4321 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 4381 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 4441 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 4501 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 4561 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 4621 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 4681 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 4741 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 4801 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 4861 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 4921 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 4981 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 5041 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 5101 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 5161 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 5221 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 5281 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 5341 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 5401 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 5461 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 5521 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 5581 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 5641 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 5701 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 5761 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 5821 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 5881 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 5941 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 6001 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 6061 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 6121 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 6181 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 6241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 6301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 6361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 6421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 6481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 6541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 6601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 6661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 6721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 6781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 6841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 6901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 6961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 7021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 7081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 7141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 7201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 7261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 7321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 7381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 7441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 7501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 7561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 7621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 7681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 7741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 7801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 7861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 7921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 7981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 8041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 8101 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc 8161 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 8221 gtatctgctc cctgcttgtg tg //