LOCUS pACP-tagm2 5543 bp DNA circular 08-APR-2011 DEFINITION Cloning vector pACP-tagm2, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pACP-tagm2 ORGANISM Cloning vector pACP-tagm2 Unclassified. REFERENCE 1 (bases 1 to 5543) 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..5543 /organism="Cloning vector pACP-tagm2" /mol_type="other DNA" promoter 251..818 /note="CMV immediate early promoter region" promoter 863..880 /note="T7 promoter (clockwise)" misc_feature 915..965 /note="multiple cloning site MCS1 (NheI-EcoRI)" gene 975..1208 /gene="ACPwt" CDS 975..1208 /gene="ACPwt" /note="ACPwt sequence 975..1208; no stop codon; possible alternative start 774" /codon_start=1 /product="wild type acyl carrier protein (ACP) from Escherichia coli (gene acpP)" /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELV MALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA" misc_feature 1209..1258 /note="multiple cloning site MCS2 (SbfI-NotI)" misc_RNA 1627..2209 /note="internal ribosome entry site (IRES) from encephalomyocarditis virus (ECMV); allows polycistronic expression of ACPwt and neoR, and required for expression of neoR" gene 2230..3033 /gene="aph(3')-II" CDS 2230..3033 /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(3435..3464) /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT) (counter-clockwise)" rep_origin complement(3788..4376) /note="pUC19 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4547..5407) /gene="bla" CDS complement(4547..5407) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 1332 a 1415 c 1395 g 1401 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 aattcaagct taccatgagc actatcgaag aacgcgttaa gaaaattatc ggcgaacagc 1021 tgggcgttaa gcaggaagaa gttaccaaca atgcttcttt cgttgaagac ctgggcgcgg 1081 attctcttga caccgttgag ctggtaatgg ctctggaaga agagtttgat actgagattc 1141 cggacgaaga agctgagaaa atcaccaccg ttcaggctgc cattgattac atcaacggcc 1201 accaggcgcc tgcaggcgga tccgcgttta aactcgaggt taattaatga gcggccgcat 1261 agataactga tccagtgtgc tggaattaat tcgctgtctg cgagggccag ctgttggggt 1321 gagtactccc tctcaaaagc gggcatgact tctgcgctaa gattgtcagt ttccaaaaac 1381 gaggaggatt tgatattcac ctggcccgcg gtgatgcctt tgagggtggc cgcgtccatc 1441 tggtcagaaa agacaatctt tttgttgtca agcttgaggt gtggcaggct tgagatctgg 1501 ccatacactt gagtgacaat gacatccact ttgcctttct ctccacaggt gtccactccc 1561 aggtccaact gcaggtcgag catgcatcta gggcggccaa ttccgcccct ctcccccccc 1621 cccttttccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg 1681 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg 1741 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg 1801 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca 1861 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct 1921 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca 1981 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa 2041 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg 2101 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg 2161 ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccgg gataattcct 2221 gcagccaata tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct 2281 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 2341 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 2401 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 2461 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 2521 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 2581 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 2641 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 2701 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 2761 aaggcgcgca tgcccgacgg cgatgatctc gtcgtgaccc atggcgatgc ctgcttgccg 2821 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 2881 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 2941 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 3001 gccttctatc gccttcttga cgagttcttc tgaggggatc aattctctag ataactgatc 3061 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc 3121 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct 3181 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 3241 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt cgagtgcatt 3301 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct 3361 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 3421 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 3481 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 3541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 3601 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 3661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 3721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 3781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 3841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 3901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 3961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 4021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 4081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 4141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 4201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 4261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 4321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 4381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 4441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 4501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 4561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 4621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 4681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 4741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 4801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 4861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 4921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 4981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 5281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 5341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 5401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 5461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 5521 cccgaaaagt gccacctgac gtc //