LOCUS pMCP-GPI 5238 bp DNA circular 17-DEC-2008 DEFINITION Cloning vector pMCP-GPI, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMCP-GPI ORGANISM Cloning vector pMCP-GPI other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5238) 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..5238 /organism="Cloning vector pMCP-GPI" /mol_type="other DNA" promoter 1..602 /note="CMV promoter region" gene 684..1094 /gene="MCP" CDS 684..1094 /gene="MCP" /note="ER signal sequence 684..758, MCP (ACP mutant (D36T,D39G) sequence 765..998, GPI anchor sequence 1005..1091" /codon_start=1 /product="mutant acyl carrier protein (ACP) from Escherichia coli (derived from gene acpP) with N-terminal ER signal and C-terminal GPI anchor peptides" /translation="MGIQGGSVLFGLLLVLAVFCHSGHSRSMSTIEERVKKIIGEQLG VKQEEVTNNASFVEDLGATSLGTVELVMALEEEFDTEIPDEEAEKITTVQAAIDYING HQAEFLENGGTSLSEKTVLLLVTPFLAAAWSLHP" sig_peptide 684..758 /gene="MCP" /note="secretory signal sequence" sig_peptide 1005..1091 /gene="MCP" /note="GPI anchor signal sequence" promoter complement(1147..1164) /note="T7 promoter" polyA_signal 1177..1560 /note="SV40 polyA" gene 1725..2585 /gene="bla" CDS 1725..2585 /gene="bla" /note="ampR (confers resistance to ampicillin) (mutant A182V)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" gene 3140..3934 /gene="aph(3')-II" CDS 3140..3934 /gene="aph(3')-II" /note="neoR, kanR, nptII (confers resistance to neomycin and kanamycin) (mutant R177S)" /codon_start=1 /product="aminoglycoside phosphotransferase from Tn5" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 3935..4384 /note="HSV-TK polyA" rep_origin 4542..5130 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" BASE COUNT 1285 a 1321 c 1319 g 1313 t ORIGIN 1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta 601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc 661 gcggtggcgg ccgcgatatc accatgggaa tccaaggagg gtctgtcctg ttcgggctgc 721 tgctcgtcct ggctgtcttc tgccattcag gtcatagcag atctatgagc actatcgaag 781 aacgcgttaa gaaaattatc ggcgaacagc tgggcgttaa gcaggaagaa gttaccaaca 841 atgcttcttt cgttgaagac ctgggcgcga cttctcttgg gaccgttgag ctggtaatgg 901 ctctggaaga agagtttgat actgagattc cggacgaaga agctgagaaa atcaccaccg 961 ttcaggctgc cattgattac atcaacggcc accaggcgga attccttgaa aatggtggga 1021 catccttatc agagaaaaca gttcttctgc tggtgactcc atttctggca gcagcctgga 1081 gccttcatcc ctaaggatcc tgactcgagg gggggcccgg taccaggtaa gtgtacccaa 1141 ttcgccctat agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag 1201 cctgaatggc gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta 1261 attgtttgtg tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac 1321 agtctgttca tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa 1381 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac 1441 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 1501 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa 1561 cgcgtaaatt gtaagcgtta ataagcagaa ggccatcctg acggatggcc tttttgcgtt 1621 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 1681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 1741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 1801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 1861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 1921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc 1981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 2041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 2101 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 2161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 2221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 2281 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 2341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 2401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 2461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 2521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 2581 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 2641 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 2701 gtgagttttc gttccactga gcgtcagaca ttgaaaaagg aagaatcctg aggcggaaag 2761 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 2941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc 3061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac 3121 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 3181 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 3241 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 3301 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg 3361 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 3421 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 3481 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 3541 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 3601 tcaggatgat ctggacgaag aacatcaggg gctcgcgcca gccgaactgt tcgccaggct 3661 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 3721 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 3781 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag aacttggcgg 3841 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 3901 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 3961 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 4021 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 4081 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca 4141 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt 4201 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca 4261 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca 4321 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc 4381 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 4441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 4501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 4561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 4621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 4681 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 4741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 4801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 4861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 4921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 4981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 5041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 5101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 5161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 5221 gataaccgta ttaccgcc //