LOCUS pCLIPf 5862 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pCLIPf, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLIPf ORGANISM Cloning vector pCLIPf Unclassified. REFERENCE 1 (bases 1 to 5862) 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..5862 /organism="Cloning vector pCLIPf" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter (clockwise)" misc_feature 915..965 /note="multiple cloning site MCS1 (NheI-EcoRI)" gene 969..1514 /gene="CLIP10f" CDS 969..1514 /gene="CLIP10f" /note="optimized for mammalian expression; no stop codon; possible alternative start 774" /codon_start=1 /product="O-6-methylguanine-DNA methyltransferase" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAV EVPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL LKVVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLG GLAVKEWLLAHEGHRLGKPGLG" misc_feature 1515..1564 /note="multiple cloning site MCS2 (SbfI-NotI)" misc_RNA 1946..2528 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of CLIP10f and neoR, and required for expression of neoR" gene 2549..3352 /gene="aph(3')-II" CDS 2549..3352 /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(3754..3783) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(4107..4695) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4866..5726) /gene="bla" CDS complement(4866..5726) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1368 a 1550 c 1509 g 1435 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 cgtcgctagc gatatcggcg cgccagcatt taaatctgta cagaccggtg 961 aattcaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca 1021 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa 1081 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc 1141 cactgatcca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg 1201 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg 1261 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagcgag agccacctgg 1321 ccgccctggt gggcaatccc gccgccaccg ccgccgtgaa caccgccctg gacggaaatc 1381 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacagcgac gtggggccct 1441 acctgggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca 1501 agcctgggct gggtcctgca ggcggatccg cgtttaaact cgaggttaat taatgagcgg 1561 ccgcttaagc cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc 1621 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag 1681 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt 1741 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg 1801 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc 1861 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat 1921 tccgcccctc tccccccccc ccttttccct cccccccccc taacgttact ggccgaagcc 1981 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt 2041 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc 2101 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc 2161 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc 2221 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg 2281 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct 2341 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat 2401 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc 2461 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc 2521 acaacccggg ataattcctg cagccaatat gggatcggcc attgaacaag atggattgca 2581 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 2641 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 2701 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 2761 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 2821 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 2881 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 2941 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 3001 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 3061 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca 3121 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 3181 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 3241 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 3301 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaggggatca 3361 attctctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 3421 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 3481 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 3541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 3601 ttaacgcgtc gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 3661 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 3721 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 3781 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 3841 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 3901 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 3961 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 4021 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4081 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4141 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 4201 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 4261 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 4321 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 4381 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 4441 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 4501 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 4561 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 4621 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 4681 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 4741 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 4801 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 4861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 4921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 4981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 5041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 5161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 5221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 5281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 5341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 5401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 5461 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 5521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 5581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 5641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 5701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 5761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 5821 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //