LOCUS Exported 7429 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Cdk2-DN-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7429) AUTHORS van den Heuvel S, Harlow E TITLE Distinct roles for cyclin-dependent kinases in cell cycle control. JOURNAL Science 1993 Dec 24;262(5142):2050-4. PUBMED 8266103 REFERENCE 2 (bases 1 to 7429) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7429 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(1158..1177) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 1223..1278 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(1277..1296) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(1636..1652) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1636..1652) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1649..1671) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1660..1676 /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(1684..1714) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1729..1750 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1867..1884) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2038..2626) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2118..2137) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2797..3657) /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 3420..3439 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3658..3762) /gene="bla" /label=AmpR promoter primer_bind 3830..3848 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3886..3908) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4008..4027 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4221..4243 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4235..4252 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4236..4252 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 4663..4808 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" CDS 4865..5659 /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(4919..4938) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5529..5548 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5853..5872) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5897..5944 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" enhancer 6237..6616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6617..6820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6771..6791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 6817..6841 /label=LNCX /note="Human CMV promoter, forward primer" intron 6932..75 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" ORIGIN 1 atgaggataa aatactctga gtccaaaccg ggcccctctg ctaaccatgt tcatgccttc 61 ttctctttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca tcattttggc 121 aaagaattcc tcgacggatc catggagaac ttccaaaagg tggaaaagat cggagagggc 181 acgtacggag ttgtgtacaa agccagaaac aagttgacgg gagaggtggt ggcgcttaag 241 aaaatccgcc tggacactga gactgagggt gtgcccagta ctgccatccg agagatctct 301 ctgcttaagg agcttaacca tcctaatatt gtcaagctgc tggatgtcat tcacacagaa 361 aataaactct acctggtttt tgaatttctg caccaagatc tcaagaaatt catggatgcc 421 tctgctctca ctggcattcc tcttcccctc atcaagagct atctgttcca gctgctccag 481 ggcctagctt tctgccattc tcatcgggtc ctccaccgag accttaaacc tcagaatctg 541 cttattaaca cagagggggc catcaagcta gcaaactttg gactagccag agcttttgga 601 gtccctgttc gtacttacac ccatgaggtg gtgaccctgt ggtaccgagc tcctgaaatc 661 ctcctgggct gcaaatatta ttccacagct gtggacatct ggagcctggg ctgcatcttt 721 gctgagatgg tgactcgccg ggccctattc cctggagatt ctgagattga ccagctcttc 781 cggatctttc ggactctggg gaccccagat gaggtggtgt ggccaggagt tacttctatg 841 cctgattaca agccaagttt ccccaagtgg gcccggcaag attttagtaa agttgtacct 901 cccctggatg aagatggacg gagcttgtta tcgcaaatgc tgcactacga ccctaacaag 961 cggatttcgg ccaaggcagc cctggctcac cctttcttcc aggatgtgac caagccagta 1021 ccccatcttc gactctatga tgtgccagat tatgcttgat agggatccgt cgaggaattc 1081 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct 1141 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc 1201 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg 1261 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag 1321 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa 1381 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc 1441 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt 1501 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc 1561 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga cggatcggcc 1621 gcaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 1681 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 1741 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 1801 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 1861 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 1921 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 1981 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 2041 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 2101 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 2161 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 2221 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 2281 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 2341 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 2401 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 2461 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 2521 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 2581 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 2641 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 2701 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 2761 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 2821 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 2881 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 2941 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 3001 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 3061 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 3121 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 3181 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 3241 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 3301 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 3361 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 3421 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 3481 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 3541 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 3601 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 3661 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 3721 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 3781 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat 3841 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 3901 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 3961 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca 4021 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa 4081 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt 4141 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag 4201 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct 4261 tcacgctgcc gcaagcactc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa 4321 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca 4381 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag 4441 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct 4501 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga 4561 tggcgcaggg gatcaagatc ctgcttcatc cccgtggccc gttgctcgcg tttgctggcg 4621 gtgtccccgg aagaaatata tttgcatgtc tttagttcta tgatgacaca aaccccgccc 4681 agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt cggggcggcg cggtcccagg 4741 tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac 4801 ccgcttaaca gcgtcaacag cgtgccgcag atctgatcaa gagacaggat gaggatcgtt 4861 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 4921 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 4981 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 5041 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 5101 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 5161 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 5221 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 5281 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 5341 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 5401 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 5461 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 5521 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 5581 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 5641 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 5701 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 5761 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 5821 ttcgcccacc ccgggagatg ggggaggcta actgaaacac ggaaggagac aataccggaa 5881 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt 5941 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 6001 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 6061 gaaggcccag ggctcgcagc caacgtcggg gcggcaagcc ctgccatagc cacgggcccc 6121 gtgggttagg gacggcggat cgcggccctc tagagagctt ggcccattgc atacgttgta 6181 tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca 6241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 6301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 6361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 6421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 6481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 6541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 6601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 6661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 6721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 6781 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat 6841 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag 6901 cctccggtcg accgatcctg agaacttcag ggtgagtttg gggacccttg attgttcttt 6961 ctttttcgct attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt 7021 tagaatggga agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca 7081 ctttctactc tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt 7141 ttcgttaaac tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc 7201 agattgtaag tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg 7261 tacttcagca cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct 7321 gcatataaat tctggctggc gtggaaatat tcttattggt agaaacaact acaccctggt 7381 catcatcctg cctttctctt tatggttaca atgatataca ctgtttgag //