LOCUS pSNAP-Cox8A 6015 bp DNA circular 13-NOV-2009 DEFINITION Cloning vector pSNAP-Cox8A, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pSNAP-Cox8A ORGANISM Cloning vector pSNAP-Cox8A other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6015) AUTHORS Brecht,A. and Muentner,K. TITLE Direct Submission JOURNAL Submitted (13-NOV-2009) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6015 /organism="Cloning vector pSNAP-Cox8A" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter" gene 930..1697 /gene="Cox8A/SNAP26m" CDS 1152..1697 /gene="Cox8A/SNAP26m" /note="Cox8A sequence 930..1136, SNAP26m sequence 1152..1697; SNAP26m optimized for mammalian expression; no stop codon" /codon_start=1 /product="human cytochrome c oxidase subunit 8A / O-6-methylguanine-DNA methyltransferase fusion" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAV EVPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL LKVVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEG GLAVKEWLLAHEGHRLGKPGLG" misc_RNA 2091..2666 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of Cox8A/SNAP26m and neoR, and required for expression of neoR" gene 2576..3505 /gene="aph(3')-II" CDS 2576..3505 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin) (mutant R177S)" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MLYMCLVEVKKTSRPPEPRGRGFPLKNTMISLPQPGIIPAANMG SAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGAL NELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSI MADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFAR LKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEE LGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 3528..3757 /gene="aph(3')-II" /note="SV40 polyA" promoter complement(3907..3936) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(4260..4848) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5019..5879) /gene="bla" CDS complement(5019..5879) /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 1388 a 1594 c 1572 g 1461 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 aagcttacca tgtccgtcct gacgccgctg ctgctgcggg 961 gcttgacagg ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttgccgc 1021 cggaggggaa gcttgggatc atggaattgg ccgttgggct tacctcctgc ttcgtgacct 1081 tcctcctgcc agcgggctgg atcctgtcac acctggagac ctacaggagg ccagaggata 1141 tcaagcttac catggacaaa gactgcgaaa tgaagcgcac caccctggat agccctctgg 1201 gcaagctgga actgtctggg tgcgaacagg gcctgcacga gatcaagctg ctgggcaaag 1261 gaacatctgc cgccgacgcc gtggaagtgc ctgccccagc cgccgtgctg ggcggaccag 1321 agccactgat gcaggccacc gcctggctca acgcctactt tcaccagcct gaggccatcg 1381 aggagttccc tgtgccagcc ctgcaccacc cagtgttcca gcaggagagc tttacccgcc 1441 aggtgctgtg gaaactgctg aaagtggtga agttcggaga ggtcatcagc taccagcagc 1501 tggccgccct ggccggcaat cccgccgcca ccgccgccgt gaaaaccgcc ctgagcggaa 1561 atcccgtgcc cattctgatc ccctgccacc gggtggtgtc tagctctggc gccgtggggg 1621 gctacgaggg cgggctcgcc gtgaaagagt ggctgctggc ccacgagggc cacagactgg 1681 gcaagcctgg gctgggtcct gcaggtatag gcgcgccagg atcctgactc gagtgaggcg 1741 gccgcataga taactgatcc agtgtgctgg aattaattcg ctgtctgcga gggccagctg 1801 ttggggtgag tactccctct caaaagcggg catgacttct gcgctaagat tgtcagtttc 1861 caaaaacgag gaggatttga tattcacctg gcccgcggtg atgcctttga gggtggccgc 1921 gtccatctgg tcagaaaaga caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga 1981 gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc 2041 cactcccagg tccaactgca ggtcgagcat gcatctaggg cggccaattc cgcccctctc 2101 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg 2161 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg 2221 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag 2281 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt 2341 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc 2401 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga 2461 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga 2521 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct 2581 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg 2641 ttttcctttg aaaaacacga tgataagctt gccacaaccc gggataattc ctgcagccaa 2701 tatgggatcg gccattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 2761 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 2821 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 2881 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 2941 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 3001 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 3061 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 3121 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga 3181 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg 3241 catgcccgac ggcgatgatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat 3301 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 3361 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 3421 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 3481 tcgccttctt gacgagttct tctgagggga tcaattctct agataactga tcataatcag 3541 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 3601 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 3661 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 3721 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtcgagtgca ttctagttgt 3781 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 3841 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 3901 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 3961 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 4021 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 4081 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 4141 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 4201 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 4261 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 4321 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 4381 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 4441 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 4501 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 4561 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 4621 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 4681 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 4741 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 4801 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 4861 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 4921 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 4981 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 5041 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 5101 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 5161 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 5221 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 5281 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 5341 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 5401 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 5461 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 5521 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 5581 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 5641 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 5701 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 5761 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 5821 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 5881 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 5941 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 6001 gtgccacctg acgtc //