LOCUS Exported 7429 bp ds-DNA circular SYN 17-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Cdk2-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 Jun 17, 2022 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(1593..1612) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 1658..1713 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(1712..1731) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(2071..2087) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2071..2087) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2084..2106) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2095..2111 /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(2119..2149) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2164..2185 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2302..2319) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2473..3061) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2553..2572) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3232..4092) /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 3855..3874 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4093..4197) /gene="bla" /label=AmpR promoter primer_bind 4265..4283 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4321..4343) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4443..4462 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4656..4678 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4670..4687 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4671..4687 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5098..5243 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" CDS 5300..6094 /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(5354..5373) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5964..5983 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6288..6307) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6332..6379 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" enhancer 6672..7051 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7052..7255 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7206..7226 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 7252..7276 /label=LNCX /note="Human CMV promoter, forward primer" intron 7367..510 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" ORIGIN 1 gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta tcaccatgga 61 ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt gtctcctctt 121 attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt gtaacgaatt 181 tttaaattca cttttgttta tttgtcagat tgtaagtact ttctctaatc actttttttt 241 caaggcaatc agggtatatt atattgtact tcagcacagt tttagagaac aattgttata 301 attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg aaatattctt 361 attggtagaa acaactacac cctggtcatc atcctgcctt tctctttatg gttacaatga 421 tatacactgt ttgagatgag gataaaatac tctgagtcca aaccgggccc ctctgctaac 481 catgttcatg ccttcttctc tttcctacag ctcctgggca acgtgctggt tgttgtgctg 541 tctcatcatt ttggcaaaga attcctcgac ggatccatgg agaacttcca aaaggtggaa 601 aagatcggag agggcacgta cggagttgtg tacaaagcca gaaacaagtt gacgggagag 661 gtggtggcgc ttaagaaaat ccgcctggac actgagactg agggtgtgcc cagtactgcc 721 atccgagaga tctctctgct taaggagctt aaccatccta atattgtcaa gctgctggat 781 gtcattcaca cagaaaataa actctacctg gtttttgaat ttctgcacca agatctcaag 841 aaattcatgg atgcctctgc tctcactggc attcctcttc ccctcatcaa gagctatctg 901 ttccagctgc tccagggcct agctttctgc cattctcatc gggtcctcca ccgagacctt 961 aaacctcaga atctgcttat taacacagag ggggccatca agctagcaga ctttggacta 1021 gccagagctt ttggagtccc tgttcgtact tacacccatg aggtggtgac cctgtggtac 1081 cgagctcctg aaatcctcct gggctgcaaa tattattcca cagctgtgga catctggagc 1141 ctgggctgca tctttgctga gatggtgact cgccgggccc tattccctgg agattctgag 1201 attgaccagc tcttccggat ctttcggact ctggggaccc cagatgaggt ggtgtggcca 1261 ggagttactt ctatgcctga ttacaagcca agtttcccca agtgggcccg gcaagatttt 1321 agtaaagttg tacctcccct ggatgaagat ggacggagct tgttatcgca aatgctgcac 1381 tacgacccta acaagcggat ttcggccaag gcagccctgg ctcacccttt cttccaggat 1441 gtgaccaagc cagtacccca tcttcgactc tatgatgtgc cagattatgc ttgataggga 1501 tccgtcgagg aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg 1561 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg 1621 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt 1681 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat 1741 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg 1801 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct 1861 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt 1921 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga 1981 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc 2041 ctcgacggat cggccgcaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 2101 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 2161 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 2221 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 2281 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 2341 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 2401 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 2461 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 2521 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 2581 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 2641 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 2701 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 2761 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 2821 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 2881 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 2941 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 3001 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 3061 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 3121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 3181 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 3241 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 3301 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 3361 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 3421 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 3481 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 3541 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 3601 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 3661 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 3721 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 3781 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 3841 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 3901 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 3961 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 4021 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 4081 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 4141 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 4201 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 4261 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 4321 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 4381 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 4441 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 4501 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 4561 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 4621 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 4681 ggccagtgcc aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag 4741 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac 4801 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg 4861 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca 4921 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg 4981 ccgccaagga tctgatggcg caggggatca agatcctgct tcatccccgt ggcccgttgc 5041 tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg 5101 acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg 5161 cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag 5221 cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatctg atcaagagac 5281 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 5341 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 5401 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 5461 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 5521 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 5581 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 5641 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 5701 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 5761 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 5821 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 5881 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 5941 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 6001 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 6061 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 6121 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 6181 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 6241 ctcatgctgg agttcttcgc ccaccccggg agatggggga ggctaactga aacacggaag 6301 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 6361 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 6421 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 6481 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aagccctgcc 6541 atagccacgg gccccgtggg ttagggacgg cggatcgcgg ccctctagag agcttggccc 6601 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt 6661 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt 6721 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 6781 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 6841 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 6901 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 6961 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 7021 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg 7081 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg 7141 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc 7201 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt 7261 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca 7321 ccgggaccga tccagcctcc ggtcgaccga tcctgagaac ttcagggtga gtttggggac 7381 ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatgga //