LOCUS pSNAPf-ADRB2 7164 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pSNAPf-ADRB2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pSNAPf-ADRB2 ORGANISM Cloning vector pSNAPf-ADRB2 Unclassified. REFERENCE 1 (bases 1 to 7164) 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..7164 /organism="Cloning vector pSNAPf-ADRB2" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter (clockwise)" gene 921..2846 /gene="SNAP26f/ADRB2" CDS 921..2846 /gene="SNAP26f/ADRB2" /note="note=5-hydroxytryptamine receptor 3A (5HT3R) signal sequence 921..992, SNAP26f sequence 1047..1592, human beta-2-adrenergic receptor (ADRB2) sequence 1605..2843; SNAP26f 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="MRLCIPQVLLALFLSMLTGPGEGSASDIGAPAFKSVQTGEFTMD KDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEVPAPAAVLGGPEPLM QATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISYSHLA ALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAVKEWLLAHEGHRL GKPGLGPAGTMGQPGNGSAFLLAPNGSHAPDHDVTQQRDEVWVVGMGIVMSLIVLAIV FGNVLVITAIAKFERLQTVTNYFITSLACADLVMGLAVVPFGAAHILMKMWTFGNFWC EFWTSIDVLCVTASIETLCVIAVDRYFAITSPFKYQSLLTKNKARVIILMVWIVSGLT SFLPIQMHWYRATHQEAINCYANETCCDFFTNQAYAIASSIVSFYVPLVIMVFVYSRV FQEAKRQLQKIDKSEGRFHVQNLSQVEQDGRTGHGLRRSSKFCLKEHKALKTLGIIMG TFTLCWLPFFIVNIVHVIQDNLIRKEVYILLNWIGYVNSGFNPLIYCRSPDFRIAFQE LLCLRRSSLKAYGNGYSSNGNTGEQSGYHVEQEKENKLLCEDLPGTEDFVGHQGTVPS DNIDSQGRNCSTNDSLL" misc_RNA 3248..3830 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of SNAP26f/ADRB2 and neoR, and required for expression of neoR" gene 3851..4654 /gene="aph(3')-II" CDS 3851..4654 /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(5056..5085) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(5409..5997) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(6168..7028) /gene="bla" CDS complement(6168..7028) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1652 a 1901 c 1856 g 1755 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 gcgctagcga tatcggcgcg ccagcattta 1021 aatctgtaca gaccggtgaa ttcaccatgg acaaagactg cgaaatgaag cgcaccaccc 1081 tggatagccc tctgggcaag ctggaactgt ctgggtgcga acagggcctg caccgtatca 1141 tcttcctggg caaaggaaca tctgccgccg acgccgtgga agtgcctgcc ccagccgccg 1201 tgctgggcgg accagagcca ctgatgcagg ccaccgcctg gctcaacgcc tactttcacc 1261 agcctgaggc catcgaggag ttccctgtgc cagccctgca ccacccagtg ttccagcagg 1321 agagctttac ccgccaggtg ctgtggaaac tgctgaaagt ggtgaagttc ggagaggtca 1381 tcagctacag ccacctggcc gccctggccg gcaatcccgc cgccaccgcc gccgtgaaaa 1441 ccgccctgag cggaaatccc gtgcccattc tgatcccctg ccaccgggtg gtgcagggcg 1501 acctggacgt ggggggctac gagggcgggc tcgccgtgaa agagtggctg ctggcccacg 1561 agggccacag actgggcaag cctgggctgg gtcctgcagg taccatgggg caacccggga 1621 acggcagcgc cttcttgctg gcacccaatg gaagccatgc gccggaccac gacgtcacgc 1681 agcaaaggga cgaggtgtgg gtggtgggca tgggcatcgt catgtctctc atcgtcctgg 1741 ccatcgtgtt tggcaatgtg ctggtcatca cagccattgc caagttcgag cgtctgcaga 1801 cggtcaccaa ctacttcatc acttcactgg cctgtgctga tctggtcatg ggcctagcag 1861 tggtgccctt tggggccgcc catattctta tgaaaatgtg gacttttggc aacttctggt 1921 gcgagttttg gacttccatt gatgtgctgt gcgtcacggc cagcattgag accctgtgcg 1981 tgatcgcagt ggatcgctac tttgccatta cttcaccttt caagtaccag agcctgctga 2041 ccaagaataa ggcccgggtg atcattctga tggtgtggat tgtgtcaggc cttacctcct 2101 tcttgcccat tcagatgcac tggtacaggg ccacccacca ggaagccatc aactgctatg 2161 ccaatgagac ctgctgtgac ttcttcacga accaagccta tgccattgcc tcttccatcg 2221 tgtccttcta cgttcccctg gtgatcatgg tcttcgtcta ctccagggtc tttcaggagg 2281 ccaaaaggca gctccagaag attgacaaat ctgagggccg cttccatgtc cagaacctta 2341 gccaggtgga gcaggatggg cggacggggc atggactccg cagatcttcc aagttctgct 2401 tgaaggagca caaagccctc aagacgttag gcatcatcat gggcactttc accctctgct 2461 ggctgccctt cttcatcgtt aacattgtgc atgtgatcca ggataacctc atccgtaagg 2521 aagtttacat cctcctaaat tggataggct atgtcaattc tggtttcaat ccccttatct 2581 actgccggag cccagatttc aggattgcct tccaggagct tctgtgcctg cgcaggtctt 2641 ctttgaaggc ctatggcaat ggctactcca gcaacggcaa cacaggggag cagagtggat 2701 atcacgtgga acaggagaaa gaaaataaac tgctgtgtga agacctccca ggcacggaag 2761 actttgtggg ccatcaaggt actgtgccta gcgataacat tgattcacaa gggaggaatt 2821 gtagtacaaa tgactcactg ctgtaaggcg cgccaggatc cctcgagtga ggcggccgca 2881 tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg 2941 tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa 3001 cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat 3061 ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg 3121 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc 3181 caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccccccc 3241 ccccttttcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt 3301 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc 3361 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag 3421 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac 3481 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc 3541 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc 3601 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca 3661 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt 3721 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg 3781 gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg ggataattcc 3841 tgcagccaat atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc 3901 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 3961 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 4021 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 4081 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 4141 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 4201 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 4261 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 4321 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 4381 caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc 4441 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 4501 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 4561 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 4621 cgccttctat cgccttcttg acgagttctt ctgaggggat caattctcta gataactgat 4681 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 4741 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 4801 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4861 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg tcgagtgcat 4921 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 4981 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 5041 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 5101 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 5161 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 5221 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 5281 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 5341 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 5401 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 5461 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 5521 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 5581 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 5641 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 5701 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 5761 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 5821 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 5881 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 5941 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 6001 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 6061 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 6121 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 6181 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 6241 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6301 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6361 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6421 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6481 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 6541 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 6601 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 6661 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 6721 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 6781 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 6841 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 6901 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 6961 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 7021 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 7081 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 7141 ccccgaaaag tgccacctga cgtc //