LOCUS pCLIP-tagm 5805 bp DNA circular 21-JAN-2010 DEFINITION Cloning vector pCLIP-tagm, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLIP-tagm ORGANISM Cloning vector pCLIP-tagm other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5805) AUTHORS Brecht,A. and Muentner,K. TITLE Direct Submission JOURNAL Submitted (21-JAN-2010) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..5805 /organism="Cloning vector pCLIP-tagm" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter" misc_feature 889..929 /note="multiple cloning site MCS1 (HindIII-NheI)" gene 930..1511 /gene="CLIP10m" CDS 930..1511 /gene="CLIP10m" /note="CLIP10m sequence 930..1475; optimized for mammalian expression; possible alternative start 774" /codon_start=1 /product="O-6-methylguanine-DNA methyltransferase" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIIFLGKGTSAADAV EVPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL LKVVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLG GLAVKEWLLAHEGHRLGKPGLGPAGIGAPGSLE" misc_feature 1477..1520 /note="multiple cloning site MCS2 (PstI-NotI)" misc_RNA 1866..2456 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of CLIP10m and neoR, and required for expression of neoR" gene 2492..3295 /gene="aph(3')-II" CDS 2492..3295 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin) (mutant R177S); possible alternative start at 2366" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSA QGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPG QDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD LDEEHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(3697..3726) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(4050..4638) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4809..5669) /gene="bla" CDS complement(4809..5669) /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 1354 a 1534 c 1498 g 1419 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 ttcgctagca tggacaaaga ctgcgaaatg aagcgcacca 961 ccctggatag ccctctgggc aagctggaac tgtctgggtg cgaacagggc ctgcacgaga 1021 tcatcttcct gggcaaagga acatctgccg ccgacgccgt ggaagtgcct gccccagccg 1081 ccgtgctggg cggaccagag ccactgatcc aggccaccgc ctggctcaac gcctactttc 1141 accagcctga ggccatcgag gagttccctg tgccagccct gcaccaccca gtgttccagc 1201 aggagagctt tacccgccag gtgctgtgga aactgctgaa agtggtgaag ttcggagagg 1261 tcatcagcga gagccacctg gccgccctgg tgggcaatcc cgccgccacc gccgccgtga 1321 acaccgccct ggacggaaat cccgtgccca ttctgatccc ctgccaccgg gtggtgcagg 1381 gcgacagcga cgtggggccc tacctgggcg ggctcgccgt gaaagagtgg ctgctggccc 1441 acgagggcca cagactgggc aagcctgggc tgggtcctgc aggtataggc gcgccaggat 1501 ccctcgagtg aggcggccgc atagataact gatccagtgt gctggaatta attcgctgtc 1561 tgcgagggcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct 1621 aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg cggtgatgcc 1681 tttgagggtg gccgcgtcca tctggtcaga aaagacaatc tttttgttgt caagcttgag 1741 gtgtggcagg cttgagatct ggccatacac ttgagtgaca atgacatcca ctttgccttt 1801 ctctccacag gtgtccactc ccaggtccaa ctgcaggtcg agcatgcatc tagggcggcc 1861 aattccgccc ctctcccccc cccccttttc cctccccccc ccctaacgtt actggccgaa 1921 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt 1981 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 2041 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 2101 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 2161 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 2221 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 2281 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg 2341 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac 2401 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt 2461 gccacaaccc gggataattc ctgcagccaa tatgggatcg gccattgaac aagatggatt 2521 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 2581 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 2641 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct 2701 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 2761 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 2821 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 2881 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 2941 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 3001 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgatgatc tcgtcgtgac 3061 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 3121 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 3181 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 3241 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagggga 3301 tcaattctct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc 3361 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 3421 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 3481 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 3541 atcttaacgc gtcgagtgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 3601 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 3661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 3721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 3781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 3841 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 3901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 3961 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4021 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4081 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4141 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 4201 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 4261 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 4321 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 4381 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 4441 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 4501 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 4561 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 4621 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 4681 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 4741 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 4801 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 4861 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 4921 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 4981 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5041 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5101 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 5161 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 5221 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 5281 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 5341 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 5401 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 5461 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 5521 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 5581 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 5641 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 5701 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 5761 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //