LOCUS pSNAP-tag(m)2 5849 bp DNA circular 11-JAN-2010 DEFINITION Cloning vector pSNAP-tag(m)2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pSNAP-tag(m)2 ORGANISM Cloning vector pSNAP-tag(m)2 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5849) AUTHORS Brecht,A., Muentner,K., Ghosh,I. and Xu,M.Q. TITLE Direct Submission JOURNAL Submitted (30-NOV-2009) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..5849 /organism="Cloning vector pSNAP-tag(m)2" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter" misc_feature 889..972 /note="multiple cloning site MCS1 (HindIII-NcoI)" gene 969..1553 /gene="SNAP26m" CDS 969..1553 /gene="SNAP26m" /note="optimized for mammalian expression; possible alternative start 774" /codon_start=1 /product="O-6-methylguanine-DNA methyltransferase" /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAV EVPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL LKVVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEG GLAVKEWLLAHEGHRLGKPGLGPAGGSAFKLEVN" misc_feature 1516..1564 /note="multiple cloning site MCS2 (PstI-NotI)" misc_RNA 1933..2515 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of CLIP10m and neoR, and required for expression of neoR" gene 2536..3339 /gene="aph(3')-II" CDS 2536..3339 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin) (mutant R177S); possible alternative start at 2410" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSA QGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPG QDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD LDEEHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(3741..3770) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(4094..4682) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4853..5713) /gene="bla" CDS complement(4853..5713) /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 1367 a 1541 c 1508 g 1433 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 tgcacgagat caagctgctg ggcaaaggaa 1081 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc 1141 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg 1201 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg 1261 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac cagcagctgg 1321 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc 1381 ccgtgcccat tctgatcccc tgccaccggg tggtgtctag ctctggcgcc gtggggggct 1441 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca 1501 agcctgggct gggtcctgca ggcggatccg cgtttaaact cgaggttaat taatgagcgg 1561 ccgcatagat aactgatcca gtgtgctgga attaattcgc tgtctgcgag ggccagctgt 1621 tggggtgagt actccctctc aaaagcgggc atgacttctg cgctaagatt gtcagtttcc 1681 aaaaacgagg aggatttgat attcacctgg cccgcggtga tgcctttgag ggtggccgcg 1741 tccatctggt cagaaaagac aatctttttg ttgtcaagct tgaggtgtgg caggcttgag 1801 atctggccat acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc 1861 actcccaggt ccaactgcag gtcgagcatg catctagggc ggccaattcc gcccctctcc 1921 ccccccccct tttccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg 1981 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag 2041 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc 2101 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg 2161 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag 2221 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca 2281 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt 2341 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc 2401 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa 2461 ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccgggata 2521 attcctgcag ccaatatggg atcggccatt gaacaagatg gattgcacgc aggttctccg 2581 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 2641 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 2701 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 2761 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 2821 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 2881 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 2941 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 3001 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 3061 aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc 3121 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 3181 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 3241 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 3301 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag gggatcaatt ctctagataa 3361 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca 3421 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 3481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 3541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtcgag 3601 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 3661 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 3721 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 3781 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 3841 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 3901 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 3961 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4021 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4501 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 4561 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 4621 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 4681 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 4741 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 4801 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 4861 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 4921 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 4981 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 5041 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 5101 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 5161 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 5221 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 5281 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 5341 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 5401 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 5461 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 5521 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 5581 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 5641 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 5701 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 5761 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 5821 catttccccg aaaagtgcca cctgacgtc //