LOCUS pCLIP-H2B 6183 bp DNA circular 17-DEC-2008 DEFINITION Cloning vector pCLIP-H2B, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLIP-H2B ORGANISM Cloning vector pCLIP-H2B other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6183) AUTHORS Brecht,A. and Muentner,K. TITLE Direct Submission JOURNAL Submitted (17-DEC-2008) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6183 /organism="Cloning vector pCLIP-H2B" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter" gene 924..1889 /gene="H2B/CLIP10m" CDS 924..1889 /gene="H2B/CLIP10m" /note="H2B sequence 924..1301, CLIP10m sequence 1308..1853; CLIP10m optimized for mammalian expression; possible alternative start 774" /codon_start=1 /product="histone H2B / O-6-methylguanine-DNA methyltransferase fusion" /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYK VLKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLL LPGELAKHAVSEGTKAITKYTSAKASMDKDCEMKRTTLDSPLGKLELSGCEQGLHEII FLGKGTSAADAVEVPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQ QESFTRQVLWKLLKVVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRV VQGDSDVGPYLGGLAVKEWLLAHEGHRLGKPGLGPAGIGAPGSLE" misc_RNA 2267..2849 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of H2B/CLIP10m and neoR, and required for expression of neoR" gene 2870..3673 /gene="aph(3')-II" CDS 2870..3673 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin) (mutant R177S); possible alternative start at 2744" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSA QGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPG QDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD LDEEHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(4075..4104) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(4428..5016) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5187..6047) /gene="bla" CDS complement(5187..6047) /gene="bla" /note="ampR (confers resistance to ampicillin) (mutant V82I,A182V)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1448 a 1645 c 1609 g 1481 t ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc 901 gagctcggat cgatatcgaa ttgatgccag agccagcgaa gtctgctccc gccccgaaaa 961 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca 1021 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca 1081 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc 1141 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca 1201 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt 1261 ccgagggtac taaggccatc accaagtaca ccagcgctaa ggctagcatg gacaaagact 1321 gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa gctggaactg tctgggtgcg 1381 aacagggcct gcacgagatc atcttcctgg gcaaaggaac atctgccgcc gacgccgtgg 1441 aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc actgatccag gccaccgcct 1501 ggctcaacgc ctactttcac cagcctgagg ccatcgagga gttccctgtg ccagccctgc 1561 accacccagt gttccagcag gagagcttta cccgccaggt gctgtggaaa ctgctgaaag 1621 tggtgaagtt cggagaggtc atcagcgaga gccacctggc cgccctggtg ggcaatcccg 1681 ccgccaccgc cgccgtgaac accgccctgg acggaaatcc cgtgcccatt ctgatcccct 1741 gccaccgggt ggtgcagggc gacagcgacg tggggcccta cctgggcggg ctcgccgtga 1801 aagagtggct gctggcccac gagggccaca gactgggcaa gcctgggctg ggtcctgcag 1861 gtataggcgc gccaggatcc ctcgagtgag gcggccgcat agataactga tccagtgtgc 1921 tggaattaat tcgctgtctg cgagggccag ctgttggggt gagtactccc tctcaaaagc 1981 gggcatgact tctgcgctaa gattgtcagt ttccaaaaac gaggaggatt tgatattcac 2041 ctggcccgcg gtgatgcctt tgagggtggc cgcgtccatc tggtcagaaa agacaatctt 2101 tttgttgtca agcttgaggt gtggcaggct tgagatctgg ccatacactt gagtgacaat 2161 gacatccact ttgcctttct ctccacaggt gtccactccc aggtccaact gcaggtcgag 2221 catgcatcta gggcggccaa ttccgcccct ctcccccccc cccttttccc tccccccccc 2281 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat 2341 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct 2401 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg 2461 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc 2521 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg 2581 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg 2641 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga 2701 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt 2761 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa 2821 aaacacgatg ataagcttgc cacaacccgg gataattcct gcagccaata tgggatcggc 2881 cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 2941 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 3001 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 3061 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 3121 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 3181 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 3241 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 3301 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 3361 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 3421 cgatgatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 3481 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 3541 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 3601 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 3661 cgagttcttc tgaggggatc aattctctag ataactgatc ataatcagcc ataccacatt 3721 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa 3781 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag 3841 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 3901 gtccaaactc atcaatgtat cttaacgcgt cgagtgcatt ctagttgtgg tttgtccaaa 3961 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4021 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4081 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4141 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4201 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4261 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 4321 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 4381 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 4441 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 4501 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 4561 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 4621 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 4681 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 4741 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 4801 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 4861 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 4921 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 4981 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5041 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5101 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5161 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5221 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 5281 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 5341 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 5401 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 5461 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 5521 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 5581 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 5641 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 5701 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 5761 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 5821 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 5881 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 5941 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6001 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6061 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6121 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6181 gtc //