LOCUS Exported 7451 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Cdc2-DN-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7451) 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 7451) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7451 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 581..605 /label=LNCX /note="Human CMV promoter, forward primer" intron 696..1268 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" primer_bind complement(2373..2392) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2438..2493 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2492..2511) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(2851..2867) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2851..2867) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2864..2886) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2875..2891 /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(2899..2929) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2944..2965 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3082..3099) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3253..3841) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3333..3352) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4012..4872) /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 4635..4654 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4873..4977) /gene="bla" /label=AmpR promoter primer_bind 5045..5063 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5101..5123) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5223..5242 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5436..5458 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5450..5467 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5451..5467 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5878..6023 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" CDS 6080..6874 /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(6134..6153) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6744..6763 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7068..7087) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7112..7159 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat 661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt 721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt 781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct 841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa 901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt 961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat 1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat 1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc 1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga 1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt 1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat 1321 tcctcgacgg atccactatg gaagattata ccaaaataga gaaaattgga gaaggtacct 1381 atggagttgt gtataagggt agacacaaaa ctacaggtca agtggtagcc atgaaaaaaa 1441 tcagactaga aagtgaagag gaaggggttc ctagtactgc aattcgggaa atttctctat 1501 taaaggaact tcgtcatcca aatatagtca gtcttcagga tgtgcttatg caggattcca 1561 ggttatatct catctttgag tttctttcca tggatctgaa gaaatacttg gattctatcc 1621 ctcctggtca gtacatggat tcttcacttg ttaagagtta tttataccaa atcctacagg 1681 ggattgtgtt ttgtcactct agaagagttc ttcacagaga cttaaaacct caaaatctct 1741 tgattgatga caaaggaaca attaaactgg ctaattttgg ccttgccaga gcttttggaa 1801 tacctatcag agtatataca catgaggtag taacactctg gtacagatct ccagaagtat 1861 tgctggggtc agctcgttac tcaactccag ttgacatttg gagtataggc accatatttg 1921 ctgaactagc aactaagaaa ccacttttcc atggggattc agaaattgat caactcttca 1981 ggattttcag agctttgggc actcccaata atgaagtgtg gccagaagtg gaatctttac 2041 aggactataa gaatacattt cccaaatgga aaccaggaag cctagcatcc catgtcaaaa 2101 acttggatga aaatggcttg gatttgctct cgaaaatgtt aatctatgat ccagccaaac 2161 gaatttctgg caaaatggca ctgaatcatc catattttaa tgatttggac aatcagatta 2221 agaagatgta tgatgtgcca gattatgctt agctttctga caaaaagttt ccatagtgga 2281 tccgtcgagg aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg 2341 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg 2401 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt 2461 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat 2521 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg 2581 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct 2641 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt 2701 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga 2761 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc 2821 ctcgacggat cggccgcaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 2881 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 2941 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3001 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3061 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 3121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 3181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 3241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 3301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 3361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 3421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 3481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 3541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 3601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 3661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 3721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 3781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 3841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 3901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 3961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 4021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 4081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 4141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 4201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 4261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 4321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 4381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 4441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 4501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 4561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 4621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 4681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 4741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 4801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 4861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 4921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 4981 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 5041 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 5101 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 5161 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 5221 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 5281 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 5341 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 5401 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 5461 ggccagtgcc aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag 5521 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac 5581 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg 5641 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca 5701 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg 5761 ccgccaagga tctgatggcg caggggatca agatcctgct tcatccccgt ggcccgttgc 5821 tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg 5881 acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg 5941 cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag 6001 cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatctg atcaagagac 6061 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 6121 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 6181 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 6241 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 6301 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 6361 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 6421 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 6481 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 6541 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 6601 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 6661 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 6721 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 6781 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 6841 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 6901 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 6961 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 7021 ctcatgctgg agttcttcgc ccaccccggg agatggggga ggctaactga aacacggaag 7081 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 7141 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 7201 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 7261 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aagccctgcc 7321 atagccacgg gccccgtggg ttagggacgg cggatcgcgg ccctctagag agcttggccc 7381 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt 7441 accgccatgt t //