LOCUS pACP-ADRB2 6824 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pACP-ADRB2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pACP-ADRB2 ORGANISM Cloning vector pACP-ADRB2 Unclassified. REFERENCE 1 (bases 1 to 6824) 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..6824 /organism="Cloning vector pACP-ADRB2" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter (clockwise)" gene 924..2498 /gene="ACPwt/ADRB2" CDS 924..2498 /gene="ACPwt/ADRB2" /note="5-hydroxytryptamine receptor 3A (5HT3R) secretory signal sequence 924..995, ACPwt sequence 1011..1244, ADRB2 sequence 1257..2495; possible alternative start 774" /codon_start=1 /product="wild type acyl carrier protein (ACP) from Escherichia coli (gene acpP) / beta-2-adrenergic receptor fusion with N-terminal 5HT3R signal peptide" /translation="MRLCIPQVLLALFLSMLTGPGEGSEFKLTMSTIEERVKKIIGEQ LGVKQEEVTNNASFVEDLGADSLDTVELVMALEEEFDTEIPDEEAEKITTVQAAIDYI NGHQAPAGTMGQPGNGSAFLLAPNGSHAPDHDVTQQRDEVWVVGMGIVMSLIVLAIVF GNVLVITAIAKFERLQTVTNYFITSLACADLVMGLAVVPFGAAHILMKMWTFGNFWCE FWTSIDVLCVTASIETLCVIAVDRYFAITSPFKYQSLLTKNKARVIILMVWIVSGLTS FLPIQMHWYRATHQEAINCYANETCCDFFTNQAYAIASSIVSFYVPLVIMVFVYSRVF QEAKRQLQKIDKSEGRFHVQNLSQVEQDGRTGHGLRRSSKFCLKEHKALKTLGIIMGT FTLCWLPFFIVNIVHVIQDNLIRKEVYILLNWIGYVNSGFNPLIYCRSPDFRIAFQEL LCLRRSSLKAYGNGYSSNGNTGEQSGYHVEQEKENKLLCEDLPGTEDFVGHQGTVPSD NIDSQGRNCSTNDSLL" misc_RNA 2908..3490 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of ACPwt/ADRB2 and neoR, and required for expression of neoR" gene 3511..4314 /gene="aph(3')-II" CDS 3511..4314 /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(4716..4745) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(5069..5657) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5828..6688) /gene="bla" CDS complement(5828..6688) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1609 a 1763 c 1735 g 1717 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 accatgcggc tctgcatccc gcaggtgctg ttggccttgt 961 tcctttccat gctgacaggg ccgggagaag gcagcgaatt caagcttacc atgagcacta 1021 tcgaagaacg cgttaagaaa attatcggcg aacagctggg cgttaagcag gaagaagtta 1081 ccaacaatgc ttctttcgtt gaagacctgg gcgcggattc tcttgacacc gttgagctgg 1141 taatggctct ggaagaagag tttgatactg agattccgga cgaagaagct gagaaaatca 1201 ccaccgttca ggctgccatt gattacatca acggccacca ggcgcctgca ggtaccatgg 1261 ggcaacccgg gaacggcagc gccttcttgc tggcacccaa tggaagccat gcgccggacc 1321 acgacgtcac gcagcaaagg gacgaggtgt gggtggtggg catgggcatc gtcatgtctc 1381 tcatcgtcct ggccatcgtg tttggcaatg tgctggtcat cacagccatt gccaagttcg 1441 agcgtctgca gacggtcacc aactacttca tcacttcact ggcctgtgct gatctggtca 1501 tgggcctagc agtggtgccc tttggggccg cccatattct tatgaaaatg tggacttttg 1561 gcaacttctg gtgcgagttt tggacttcca ttgatgtgct gtgcgtcacg gccagcattg 1621 agaccctgtg cgtgatcgca gtggatcgct actttgccat tacttcacct ttcaagtacc 1681 agagcctgct gaccaagaat aaggcccggg tgatcattct gatggtgtgg attgtgtcag 1741 gccttacctc cttcttgccc attcagatgc actggtacag ggccacccac caggaagcca 1801 tcaactgcta tgccaatgag acctgctgtg acttcttcac gaaccaagcc tatgccattg 1861 cctcttccat cgtgtccttc tacgttcccc tggtgatcat ggtcttcgtc tactccaggg 1921 tctttcagga ggccaaaagg cagctccaga agattgacaa atctgagggc cgcttccatg 1981 tccagaacct tagccaggtg gagcaggatg ggcggacggg gcatggactc cgcagatctt 2041 ccaagttctg cttgaaggag cacaaagccc tcaagacgtt aggcatcatc atgggcactt 2101 tcaccctctg ctggctgccc ttcttcatcg ttaacattgt gcatgtgatc caggataacc 2161 tcatccgtaa ggaagtttac atcctcctaa attggatagg ctatgtcaat tctggtttca 2221 atccccttat ctactgccgg agcccagatt tcaggattgc cttccaggag cttctgtgcc 2281 tgcgcaggtc ttctttgaag gcctatggca atggctactc cagcaacggc aacacagggg 2341 agcagagtgg atatcacgtg gaacaggaga aagaaaataa actgctgtgt gaagacctcc 2401 caggcacgga agactttgtg ggccatcaag gtactgtgcc tagcgataac attgattcac 2461 aagggaggaa ttgtagtaca aatgactcac tgctgtaagg cgcgccagga tccctcgagg 2521 ttaattaatg agcggccgca tagataactg atccagtgtg ctggaattaa ttcgctgtct 2581 gcgagggcca gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta 2641 agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc ggtgatgcct 2701 ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc aagcttgagg 2761 tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac tttgcctttc 2821 tctccacagg tgtccactcc caggtccaac tgcaggtcga gcatgcatct agggcggcca 2881 attccgcccc tctccccccc ccccttttcc ctcccccccc cctaacgtta ctggccgaag 2941 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc 3001 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg 3061 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc 3121 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc 3181 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa 3241 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct 3301 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg 3361 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg 3421 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg 3481 ccacaacccg ggataattcc tgcagccaat atgggatcgg ccattgaaca agatggattg 3541 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 3601 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 3661 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 3721 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 3781 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 3841 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 3901 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 3961 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 4021 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc 4081 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 4141 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 4201 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 4261 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaggggat 4321 caattctcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct 4381 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 4441 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4501 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 4561 tcttaacgcg tcgagtgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 4621 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 4681 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 4741 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 4801 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 4861 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 4921 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 4981 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 5041 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 5101 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 5161 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 5221 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 5281 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 5341 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 5401 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 5461 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 5521 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 5581 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 5641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 6001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6781 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //