LOCUS pSNAPf-H2B 6236 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pSNAPf-H2B, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pSNAPf-H2B ORGANISM Cloning vector pSNAPf-H2B Unclassified. REFERENCE 1 (bases 1 to 6236) 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..6236 /organism="Cloning vector pSNAPf-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/SNAP26f" /product="histone H2B / O-6-methylguanine-DNA methyltransferase fusion" /note="note=H2B sequence 924..1301, SNAP26f sequence 1356..1901; SNAP26f optimized for mammalian expression; possible alternative start 774" misc_RNA 2320..2902 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of H2B/SNAP26f and neoR, and required for expression of neoR" gene 2923..3726 /gene="aph(3')-II" CDS 2923..3726 /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(4128..4157) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(4481..5069) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5240..6100) /gene="bla" CDS complement(5240..6100) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1465 a 1655 c 1618 g 1498 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 tgatgcaggc caccgcctgg ctcaacgcct 1561 actttcacca gcctgaggcc atcgaggagt tccctgtgcc agccctgcac cacccagtgt 1621 tccagcagga gagctttacc cgccaggtgc tgtggaaact gctgaaagtg gtgaagttcg 1681 gagaggtcat cagctacagc cacctggccg ccctggccgg caatcccgcc gccaccgccg 1741 ccgtgaaaac cgccctgagc ggaaatcccg tgcccattct gatcccctgc caccgggtgg 1801 tgcagggcga cctggacgtg gggggctacg agggcgggct cgccgtgaaa gagtggctgc 1861 tggcccacga gggccacaga ctgggcaagc ctgggctggg tcctgcaggc ggatccgcgt 1921 ttaaactcga ggttaattaa tgagcggccg catagataac tgatccagtg tgctggaatt 1981 aattcgctgt ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa agcgggcatg 2041 acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc 2101 gcggtgatgc ctttgagggt ggccgcgtcc atctggtcag aaaagacaat ctttttgttg 2161 tcaagcttga ggtgtggcag gcttgagatc tggccataca cttgagtgac aatgacatcc 2221 actttgcctt tctctccaca ggtgtccact cccaggtcca actgcaggtc gagcatgcat 2281 ctagggcggc caattccgcc cctctccccc cccccctttt ccctcccccc cccctaacgt 2341 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac 2401 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag 2461 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa 2521 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag 2581 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga 2641 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag 2701 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc 2761 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag 2821 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg 2881 atgataagct tgccacaacc cgggataatt cctgcagcca atatgggatc ggccattgaa 2941 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3001 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3061 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3121 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3181 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3241 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3301 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3361 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3421 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat 3481 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3541 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3601 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3661 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3721 ttctgagggg atcaattctc tagataactg atcataatca gccataccac atttgtagag 3781 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat 3841 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc 3901 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 3961 ctcatcaatg tatcttaacg cgtcgagtgc attctagttg tggtttgtcc aaactcatca 4021 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 4081 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4141 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4201 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 4381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 4441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 4501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 4561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 4621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 4681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 4741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 4801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 4861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 4921 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 4981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 5041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 5641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 5701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 5761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 5821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 5881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 5941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //