LOCUS Exported 7444 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Cdk4-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7444) 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 7444) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7444 /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(2366..2385) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2431..2486 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2485..2504) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(2844..2860) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2844..2860) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2857..2879) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2868..2884 /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(2892..2922) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2937..2958 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3075..3092) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3246..3834) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3326..3345) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4005..4865) /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 4628..4647 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4866..4970) /gene="bla" /label=AmpR promoter primer_bind 5038..5056 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5094..5116) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5216..5235 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5429..5451 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5443..5460 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5444..5460 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5871..6016 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" CDS 6073..6867 /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(6127..6146) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6737..6756 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7061..7080) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7105..7152 /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 atccaccatg gctacctctc gatatgagcc agtggctgaa attggtgtcg 1381 gtgcctatgg gacagtgtac aaggcccgtg atccccacag tggccacttt gtggccctca 1441 agagtgtgag agtccccaat ggaggaggag gtggaggagg ccttcccatc agcacagttc 1501 gtgaggtggc tttactgagg cgactggagg cttttgagca tcccaatgtt gtccggctga 1561 tggacgtctg tgccacatcc cgaactgacc gggagatcaa ggtaaccctg gtgtttgagc 1621 atgtagacca ggacctaagg acatatctgg acaaggcacc cccaccaggc ttgccagccg 1681 aaacgatcaa ggatctgatg cgccagtttc taagaggcct agatttcctt catgccaatt 1741 gcatcgttca ccgagatctg aagccagaga acattctggt gacaagtggt ggaacagtca 1801 agctggctga ctttggcctg gccagaatct acagctacca gatggcactt acacccgtgg 1861 ttgttacact ctggtaccga gctcccgaag ttcttctgca gtccacatat gcaacacctg 1921 tggacatgtg gagtgttggc tgtatctttg cagagatgtt tcgtcgaaag cctctcttct 1981 gtggaaactc tgaagccgac cagttgggca aaatctttga cctgattggg ctgcctccag 2041 aggatgactg gcctcgagat gtatccctgc cccgtggagc ctttcccccc agagggcccc 2101 gcccagtgca gtcggtggta cctgagatgg aggagtcggg agcacagctg ctgctggaaa 2161 tgctgacttt taacccacac aagcgaatct ctgcctttcg agctctgcag cactcttatc 2221 tacataagga tgaaggtaat ccggagtatg atgtgccaga ttatgcttga ggatccgtcg 2281 aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg 2341 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca 2401 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag 2461 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa 2521 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca 2581 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt 2641 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt 2701 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc 2761 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacg 2821 gatcggccgc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc 2881 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 2941 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 3001 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 3061 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 3121 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 3181 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 3241 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 3301 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 3361 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 3421 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 3481 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 3541 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 3601 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 3661 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 3721 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 3781 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 3841 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 3901 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 3961 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 4021 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 4081 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 4141 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 4201 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 4261 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 4321 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 4381 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 4441 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 4501 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 4561 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 4621 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 4681 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 4741 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 4801 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 4861 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 4921 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 4981 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 5041 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 5101 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 5161 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 5221 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta 5281 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg 5341 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg 5401 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt 5461 gccaagcttc acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg aagcggaaca 5521 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta 5581 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat 5641 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg 5701 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa 5761 ggatctgatg gcgcagggga tcaagatcct gcttcatccc cgtggcccgt tgctcgcgtt 5821 tgctggcggt gtccccggaa gaaatatatt tgcatgtctt tagttctatg atgacacaaa 5881 ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg 5941 gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct 6001 gcagcgaccc gcttaacagc gtcaacagcg tgccgcagat ctgatcaaga gacaggatga 6061 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 6121 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 6181 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 6241 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 6301 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 6361 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 6421 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 6481 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 6541 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 6601 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 6661 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 6721 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 6781 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 6841 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 6901 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 6961 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 7021 tggagttctt cgcccacccc gggagatggg ggaggctaac tgaaacacgg aaggagacaa 7081 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg 7141 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac 7201 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa 7261 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaagccct gccatagcca 7321 cgggccccgt gggttaggga cggcggatcg cggccctcta gagagcttgg cccattgcat 7381 acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac attaccgcca 7441 tgtt 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