LOCUS pCLIPf-H2B 6249 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pCLIPf-H2B, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLIPf-H2B ORGANISM Cloning vector pCLIPf-H2B Unclassified. REFERENCE 1 (bases 1 to 6249) AUTHORS Sun,L., Provost,C., Ghosh,I. and Xu,M.Q. TITLE Direct Submission JOURNAL Submitted (08-APR-2011) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6249 /organism="Cloning vector pCLIPf-H2B" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter (clockwise)" gene 924..1940 /gene="H2B/CLIP10f" CDS 924..1940 /gene="H2B/CLIP10f" /note="H2B sequence 924..1301, CLIP10f sequence 1356..1901; CLIP10f optimized for mammalian expression; possible alternative start 774" /codon_start=1 /product="histone H2B / O-6-methylguanine-DNA methyltransferase fusion" /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYK VLKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLL LPGELAKHAVSEGTKAITKYTSAKASDIGAPAFKSVQTGEFTMDKDCEMKRTTLDSPL GKLELSGCEQGLHRIIFLGKGTSAADAVEVPAPAAVLGGPEPLIQATAWLNAYFHQPE AIEEFPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISESHLAALVGNPAATAAVNT ALDGNPVPILIPCHRVVQGDSDVGPYLGGLAVKEWLLAHEGHRLGKPGLGPAGGSAFK LEVN" misc_RNA 2333..2915 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of H2B/CLIP10f and neoR, and required for expression of neoR" gene 2936..3739 /gene="aph(3')-II" CDS 2936..3739 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin)" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSA QGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPG QDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD LDEEHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(4141..4170) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(4494..5082) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5253..6113) /gene="bla" CDS complement(5253..6113) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1466 a 1662 c 1621 g 1500 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 ggctagcgat atcggcgcgc 1321 cagcatttaa atctgtacag accggtgaat tcaccatgga caaagactgc gaaatgaagc 1381 gcaccaccct ggatagccct ctgggcaagc tggaactgtc tgggtgcgaa cagggcctgc 1441 accgtatcat cttcctgggc aaaggaacat ctgccgccga cgccgtggaa gtgcctgccc 1501 cagccgccgt gctgggcgga ccagagccac tgatccaggc caccgcctgg ctcaacgcct 1561 actttcacca gcctgaggcc atcgaggagt tccctgtgcc agccctgcac cacccagtgt 1621 tccagcagga gagctttacc cgccaggtgc tgtggaaact gctgaaagtg gtgaagttcg 1681 gagaggtcat cagcgagagc cacctggccg ccctggtggg caatcccgcc gccaccgccg 1741 ccgtgaacac cgccctggac ggaaatcccg tgcccattct gatcccctgc caccgggtgg 1801 tgcagggcga cagcgacgtg gggccctacc tgggcgggct cgccgtgaaa gagtggctgc 1861 tggcccacga gggccacaga ctgggcaagc ctgggctggg tcctgcaggc ggatccgcgt 1921 ttaaactcga ggttaattaa tgagcggccg cttaagccgg ccgcatagat aactgatcca 1981 gtgtgctgga attaattcgc tgtctgcgag ggccagctgt tggggtgagt actccctctc 2041 aaaagcgggc atgacttctg cgctaagatt gtcagtttcc aaaaacgagg aggatttgat 2101 attcacctgg cccgcggtga tgcctttgag ggtggccgcg tccatctggt cagaaaagac 2161 aatctttttg ttgtcaagct tgaggtgtgg caggcttgag atctggccat acacttgagt 2221 gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcag 2281 gtcgagcatg catctagggc ggccaattcc gcccctctcc ccccccccct tttccctccc 2341 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata 2401 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg 2461 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt 2521 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag 2581 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc 2641 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga 2701 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg 2761 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat 2821 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc 2881 tttgaaaaac acgatgataa gcttgccaca acccgggata attcctgcag ccaatatggg 2941 atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 3001 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 3061 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 3121 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 3181 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 3241 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 3301 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 3361 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 3421 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 3481 cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 3541 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 3601 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 3661 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 3721 tcttgacgag ttcttctgag gggatcaatt ctctagataa ctgatcataa tcagccatac 3781 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 3841 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 3901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 3961 tggtttgtcc aaactcatca atgtatctta acgcgtcgag tgcattctag ttgtggtttg 4021 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 4081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 4141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 4201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 4261 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 4321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 4381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 4441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 4501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 4561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 4621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 4681 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 4741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 4801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 4861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 4921 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 4981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 5041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 5101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 5161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 5221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 5281 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 5341 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 5401 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 5461 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 5521 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 5581 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 5641 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 5701 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 5761 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 5821 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 5881 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 5941 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6001 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 6061 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 6121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 6181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 6241 cctgacgtc //