LOCUS pCLIPf-Cox8A 6077 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pCLIPf-Cox8A, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pCLIPf-Cox8A ORGANISM Cloning vector pCLIPf-Cox8A Unclassified. REFERENCE 1 (bases 1 to 6077) 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..6077 /organism="Cloning vector pCLIPf-Cox8A" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter (clockwise)" gene 929..1768 /gene="Cox8A/CLIP10f" CDS 929..1768 /gene="Cox8A/CLIP10f" /note="Cox8A sequence 929..1135, CLIP10f sequence 1184..1729" /codon_start=1 /product="human cytochrome c oxidase subunit 8A / O-6-methylguanine-DNA methyltransferase fusion" /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLPPEGKLGIMELAVGL TSCFVTFLLPAGWILSHLETYRRPEDIGAPAFKSVQTGEFTMDKDCEMKRTTLDSPLG KLELSGCEQGLHRIIFLGKGTSAADAVEVPAPAAVLGGPEPLIQATAWLNAYFHQPEA IEEFPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISESHLAALVGNPAATAAVNTA LDGNPVPILIPCHRVVQGDSDVGPYLGGLAVKEWLLAHEGHRLGKPGLGPAGGSAFKL EVN" misc_RNA 2161..2743 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of Cox8A/CLIP10f and neoR, and required for expression of neoR" gene 2764..3567 /gene="aph(3')-II" CDS 2764..3567 /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(3969..3998) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(4322..4910) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5081..5941) /gene="bla" CDS complement(5081..5941) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1399 a 1621 c 1579 g 1478 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 cgatggccgc agcttaccat gtccgtcctg acgccgctgc tgctgcgggg 961 cttgacaggc tcggcccggc ggctcccagt gccgcgcgcc aagatccatt cgttgccgcc 1021 ggaggggaag cttgggatca tggaattggc cgttgggctt acctcctgct tcgtgacctt 1081 cctcctgcca gcgggctgga tcctgtcaca cctggagacc tacaggaggc cagaggatat 1141 cggcgcgcca gcatttaaat ctgtacagac cggtgaattc accatggaca aagactgcga 1201 aatgaagcgc accaccctgg atagccctct gggcaagctg gaactgtctg ggtgcgaaca 1261 gggcctgcac cgtatcatct tcctgggcaa aggaacatct gccgccgacg ccgtggaagt 1321 gcctgcccca gccgccgtgc tgggcggacc agagccactg atccaggcca ccgcctggct 1381 caacgcctac tttcaccagc ctgaggccat cgaggagttc cctgtgccag ccctgcacca 1441 cccagtgttc cagcaggaga gctttacccg ccaggtgctg tggaaactgc tgaaagtggt 1501 gaagttcgga gaggtcatca gcgagagcca cctggccgcc ctggtgggca atcccgccgc 1561 caccgccgcc gtgaacaccg ccctggacgg aaatcccgtg cccattctga tcccctgcca 1621 ccgggtggtg cagggcgaca gcgacgtggg gccctacctg ggcgggctcg ccgtgaaaga 1681 gtggctgctg gcccacgagg gccacagact gggcaagcct gggctgggtc ctgcaggcgg 1741 atccgcgttt aaactcgagg ttaattaatg agcggccgct taagccggcc gcatagataa 1801 ctgatccagt gtgctggaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac 1861 tccctctcaa aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag 1921 gatttgatat tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca 1981 gaaaagacaa tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac 2041 acttgagtga caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc 2101 aactgcaggt cgagcatgca tctagggcgg ccaattccgc ccctctcccc cccccccttt 2161 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt 2221 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc 2281 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca 2341 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac 2401 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg 2461 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt 2521 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct 2581 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg 2641 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt 2701 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggataat tcctgcagcc 2761 aatatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 2821 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 2881 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 2941 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 3001 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 3061 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 3121 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 3181 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 3241 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 3301 cgcatgcccg acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 3361 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 3421 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 3481 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 3541 tatcgccttc ttgacgagtt cttctgaggg gatcaattct ctagataact gatcataatc 3601 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg 3661 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat 3721 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 3781 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg cattctagtt 3841 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 3901 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 3961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4081 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4141 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4201 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4741 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 4801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 4861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 4921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 4981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 5041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 5101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 5161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 5401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 5461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 5521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 5581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 5641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 5701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 5761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 5821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 5881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 5941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 6001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 6061 aagtgccacc tgacgtc //