LOCUS pSNAPf-Cox8A 6064 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pSNAPf-Cox8A, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pSNAPf-Cox8A ORGANISM Cloning vector pSNAPf-Cox8A Unclassified. REFERENCE 1 (bases 1 to 6064) 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..6064 /organism="Cloning vector pSNAPf-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/SNAP26f" /product="human cytochrome c oxidase subunit 8A / O-6-methylguanine-DNA methyltransferase fusion" /note="Cox8A sequence 929..1135, SNAP26f sequence 1184..1729; SNAP26f optimized for mammalian expression" misc_RNA 2148..2730 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of Cox8A/SNAP26f and neoR, and required for expression of neoR" gene 2751..3554 /gene="aph(3')-II" CDS 2751..3554 /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(3956..3985) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(4309..4897) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5068..5928) /gene="bla" CDS complement(5068..5928) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1398 a 1614 c 1576 g 1476 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 atgcaggcca ccgcctggct 1381 caacgcctac tttcaccagc ctgaggccat cgaggagttc cctgtgccag ccctgcacca 1441 cccagtgttc cagcaggaga gctttacccg ccaggtgctg tggaaactgc tgaaagtggt 1501 gaagttcgga gaggtcatca gctacagcca cctggccgcc ctggccggca atcccgccgc 1561 caccgccgcc gtgaaaaccg ccctgagcgg aaatcccgtg cccattctga tcccctgcca 1621 ccgggtggtg cagggcgacc tggacgtggg gggctacgag ggcgggctcg ccgtgaaaga 1681 gtggctgctg gcccacgagg gccacagact gggcaagcct gggctgggtc ctgcaggcgg 1741 atccgcgttt aaactcgagg ttaattaatg agcggccgca tagataactg atccagtgtg 1801 ctggaattaa ttcgctgtct gcgagggcca gctgttgggg tgagtactcc ctctcaaaag 1861 cgggcatgac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca 1921 cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct 1981 ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg gccatacact tgagtgacaa 2041 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac tgcaggtcga 2101 gcatgcatct agggcggcca attccgcccc tctccccccc ccccttttcc ctcccccccc 2161 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta 2221 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc 2281 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat 2341 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc 2401 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt 2461 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt 2521 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag 2581 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt 2641 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga 2701 aaaacacgat gataagcttg ccacaacccg ggataattcc tgcagccaat atgggatcgg 2761 ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 2821 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 2881 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 2941 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3001 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3061 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 3121 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 3181 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 3241 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 3301 gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 3361 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 3421 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 3481 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 3541 acgagttctt ctgaggggat caattctcta gataactgat cataatcagc cataccacat 3601 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata 3661 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa 3721 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 3781 tgtccaaact catcaatgta tcttaacgcg tcgagtgcat tctagttgtg gtttgtccaa 3841 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 3901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 3961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 4381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 4441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 4501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 4561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 4621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 4681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 4741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 4801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 4861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 4921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 4981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 5041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 5101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 5161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 5281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 5341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 5401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 5461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 5521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 5581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 5641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 5701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 5761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 5821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 5881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 5941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6061 cgtc //