LOCUS pSNAP-tagm 5802 bp DNA circular 30-NOV-2010 DEFINITION Cloning vector pSNAP-tagm, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pSNAP-tagm ORGANISM Cloning vector pSNAP-tagm other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5802) AUTHORS Brecht,A. and Muentner,K. TITLE Direct Submission JOURNAL Submitted (30-NOV-2010) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..5802 /organism="Cloning vector pSNAP-tagm" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter" misc_feature 909..923 /note="multiple cloning site MCS1 (ClaI-EcoRI)" gene 927..1508 /gene="SNAP26m" CDS 927..1508 /gene="SNAP26m" /note="SNAP26m sequence 927..1472, vector-encoded sequence 1473..1508 (encoding C-terminal residues PAGIGAPGSLE); optimized for mammalian expression; possible alternative start 774" /codon_start=1 /product="O-6-methylguanine-DNA methyltransferase" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAV EVPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL LKVVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEG GLAVKEWLLAHEGHRLGKPGLGPAGIGAPGSLE" misc_feature 1473..1517 /note="multiple cloning site MCS2 (SbfI-NotI)" misc_RNA 1886..2468 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of SNAP26m and neoR, and required for expression of neoR" gene 2489..3292 /gene="aph(3')-II" CDS 2489..3292 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin) (mutant R177S); possible alternative start at 2363" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSA QGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPG QDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD LDEEHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(3694..3723) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(4047..4635) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4806..5666) /gene="bla" CDS complement(4806..5666) /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 1531 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 ttcaccatgg acaaagactg cgaaatgaag cgcaccaccc 961 tggatagccc tctgggcaag ctggaactgt ctgggtgcga acagggcctg cacgagatca 1021 agctgctggg caaaggaaca tctgccgccg acgccgtgga agtgcctgcc ccagccgccg 1081 tgctgggcgg accagagcca ctgatgcagg ccaccgcctg gctcaacgcc tactttcacc 1141 agcctgaggc catcgaggag ttccctgtgc cagccctgca ccacccagtg ttccagcagg 1201 agagctttac ccgccaggtg ctgtggaaac tgctgaaagt ggtgaagttc ggagaggtca 1261 tcagctacca gcagctggcc gccctggccg gcaatcccgc cgccaccgcc gccgtgaaaa 1321 ccgccctgag cggaaatccc gtgcccattc tgatcccctg ccaccgggtg gtgtctagct 1381 ctggcgccgt ggggggctac gagggcgggc tcgccgtgaa agagtggctg ctggcccacg 1441 agggccacag actgggcaag cctgggctgg gtcctgcagg tataggcgcg ccaggatccc 1501 tcgagtgagg cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc 1561 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag 1621 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt 1681 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg 1741 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc 1801 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat 1861 tccgcccctc tccccccccc ccttttccct cccccccccc taacgttact ggccgaagcc 1921 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt 1981 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc 2041 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc 2101 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc 2161 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg 2221 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct 2281 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat 2341 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc 2401 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc 2461 acaacccggg ataattcctg cagccaatat gggatcggcc attgaacaag atggattgca 2521 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 2581 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 2641 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 2701 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 2761 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 2821 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 2881 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 2941 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 3001 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca 3061 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 3121 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 3181 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 3241 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaggggatca 3301 attctctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 3361 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 3421 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 3481 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 3541 ttaacgcgtc gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 3601 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 3661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 3721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 3781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 3841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 3901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 3961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 4141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 4201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 4261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 4321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 4381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 4441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 4501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 4561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 4621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 4681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 4741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 4801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 4861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 4921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 4981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 5101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 5161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 5221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 5281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 5341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 5401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 5461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 5521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 5581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 5641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 5701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 5761 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //