LOCUS pSNAP-ADRB2 7122 bp DNA circular 17-DEC-2008 DEFINITION Cloning vector pSNAP-ADRB2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pSNAP-ADRB2 ORGANISM Cloning vector pSNAP-ADRB2 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 7122) AUTHORS Brecht,A. and Muentner,K. TITLE Direct Submission JOURNAL Submitted (17-DEC-2008) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..7122 /organism="Cloning vector pSNAP-ADRB2" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter" gene 921..2804 /gene="SNAP26m/ADBR2" CDS 921..2804 /gene="SNAP26m/ADBR2" /note="5HT3R signal sequence 921..992, SNAP26m sequence 1005..1550, human beta-2-adrenergic receptor (ADBR2) sequence 1563..2801; SNAP26m optimized for mammalian expression; possible alternative start 774" /codon_start=1 /product="O-6-methylguanine-DNA methyltransferase / beta-2-adrenergic receptor fusion with N-terminal 5HT3R signal peptide" /translation="MRLCIPQVLLALFLSMLTGPGEGSRKLTMDKDCEMKRTTLDSPL GKLELSGCEQGLHEIKLLGKGTSAADAVEVPAPAAVLGGPEPLMQATAWLNAYFHQPE AIEEFPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISYQQLAALAGNPAATAAVKT ALSGNPVPILIPCHRVVSSSGAVGGYEGGLAVKEWLLAHEGHRLGKPGLGPAGTMGQP GNGSAFLLAPNGSHAPDHDVTQQRDEVWVVGMGIVMSLIVLAIVFGNVLVITAIAKFE RLQTVTNYFITSLACADLVMGLAVVPFGAAHILMKMWTFGNFWCEFWTSIDVLCVTAS IETLCVIAVDRYFAITSPFKYQSLLTKNKARVIILMVWIVSGLTSFLPIQMHWYRATH QEAINCYANETCCDFFTNQAYAIASSIVSFYVPLVIMVFVYSRVFQEAKRQLQKIDKS EGRFHVQNLSQVEQDGRTGHGLRRSSKFCLKEHKALKTLGIIMGTFTLCWLPFFIVNI VHVIQDNLIRKEVYILLNWIGYVNSGFNPLIYCRSPDFRIAFQELLCLRRSSLKAYGN GYSSNGNTGEQSGYHVEQEKENKLLCEDLPGTEDFVGHQGTVPSDNIDSQGRNCSTND SLL" sig_peptide 921..992 /gene="SNAP26m/ADBR2" /note="5-hydroxytryptamine receptor 3A (5HT3R) signal sequence" misc_RNA 3206..3788 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of SNAP26m/ADBR2 and neoR, and required for expression of neoR" gene 3809..4612 /gene="aph(3')-II" CDS 3809..4612 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin) (mutant R177S); possible alternative start at 3683" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSA QGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPG QDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDD LDEEHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVA DRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(5014..5043) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)" rep_origin complement(5367..5955) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(6126..6986) /gene="bla" CDS complement(6126..6986) /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 1641 a 1888 c 1848 g 1745 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 cgatatcacc atgcggctct gcatcccgca ggtgctgttg gccttgttcc 961 tttccatgct gacagggccg ggagaaggca gccggaagct taccatggac aaagactgcg 1021 aaatgaagcg caccaccctg gatagccctc tgggcaagct ggaactgtct gggtgcgaac 1081 agggcctgca cgagatcaag ctgctgggca aaggaacatc tgccgccgac gccgtggaag 1141 tgcctgcccc agccgccgtg ctgggcggac cagagccact gatgcaggcc accgcctggc 1201 tcaacgccta ctttcaccag cctgaggcca tcgaggagtt ccctgtgcca gccctgcacc 1261 acccagtgtt ccagcaggag agctttaccc gccaggtgct gtggaaactg ctgaaagtgg 1321 tgaagttcgg agaggtcatc agctaccagc agctggccgc cctggccggc aatcccgccg 1381 ccaccgccgc cgtgaaaacc gccctgagcg gaaatcccgt gcccattctg atcccctgcc 1441 accgggtggt gtctagctct ggcgccgtgg ggggctacga gggcgggctc gccgtgaaag 1501 agtggctgct ggcccacgag ggccacagac tgggcaagcc tgggctgggt cctgcaggta 1561 ccatggggca acccgggaac ggcagcgcct tcttgctggc acccaatgga agccatgcgc 1621 cggaccacga cgtcacgcag caaagggacg aggtgtgggt ggtgggcatg ggcatcgtca 1681 tgtctctcat cgtcctggcc atcgtgtttg gcaatgtgct ggtcatcaca gccattgcca 1741 agttcgagcg tctgcagacg gtcaccaact acttcatcac ttcactggcc tgtgctgatc 1801 tggtcatggg cctagcagtg gtgccctttg gggccgccca tattcttatg aaaatgtgga 1861 cttttggcaa cttctggtgc gagttttgga cttccattga tgtgctgtgc gtcacggcca 1921 gcattgagac cctgtgcgtg atcgcagtgg atcgctactt tgccattact tcacctttca 1981 agtaccagag cctgctgacc aagaataagg cccgggtgat cattctgatg gtgtggattg 2041 tgtcaggcct tacctccttc ttgcccattc agatgcactg gtacagggcc acccaccagg 2101 aagccatcaa ctgctatgcc aatgagacct gctgtgactt cttcacgaac caagcctatg 2161 ccattgcctc ttccatcgtg tccttctacg ttcccctggt gatcatggtc ttcgtctact 2221 ccagggtctt tcaggaggcc aaaaggcagc tccagaagat tgacaaatct gagggccgct 2281 tccatgtcca gaaccttagc caggtggagc aggatgggcg gacggggcat ggactccgca 2341 gatcttccaa gttctgcttg aaggagcaca aagccctcaa gacgttaggc atcatcatgg 2401 gcactttcac cctctgctgg ctgcccttct tcatcgttaa cattgtgcat gtgatccagg 2461 ataacctcat ccgtaaggaa gtttacatcc tcctaaattg gataggctat gtcaattctg 2521 gtttcaatcc ccttatctac tgccggagcc cagatttcag gattgccttc caggagcttc 2581 tgtgcctgcg caggtcttct ttgaaggcct atggcaatgg ctactccagc aacggcaaca 2641 caggggagca gagtggatat cacgtggaac aggagaaaga aaataaactg ctgtgtgaag 2701 acctcccagg cacggaagac tttgtgggcc atcaaggtac tgtgcctagc gataacattg 2761 attcacaagg gaggaattgt agtacaaatg actcactgct gtaaggcgcg ccaggatccc 2821 tcgagtgagg cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc 2881 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag 2941 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt 3001 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg 3061 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc 3121 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat 3181 tccgcccctc tccccccccc ccttttccct cccccccccc taacgttact ggccgaagcc 3241 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt 3301 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc 3361 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc 3421 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc 3481 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg 3541 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct 3601 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat 3661 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc 3721 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc 3781 acaacccggg ataattcctg cagccaatat gggatcggcc attgaacaag atggattgca 3841 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 3901 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 3961 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 4021 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 4081 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 4141 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 4201 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 4261 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 4321 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca 4381 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 4441 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 4501 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 4561 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaggggatca 4621 attctctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 4681 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 4741 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4861 ttaacgcgtc gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 4921 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 4981 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 5041 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 5101 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 5161 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 5221 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 5281 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 5341 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 5401 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 5461 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 5521 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 5581 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 5641 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 5701 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 5761 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 5821 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5881 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 5941 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 6001 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 6061 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 6121 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 6181 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 6241 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 6301 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 6361 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 6421 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 6481 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 6541 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 6601 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 6661 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 6721 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 6781 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 6841 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 6901 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 6961 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 7021 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 7081 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //