LOCUS 40924_14795 3518 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pDM276, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3518) AUTHORS Veltman DM, Akar G, Bosgraaf L, Van Haastert PJ. TITLE A new set of small, extrachromosomal expression vectors for Dictyostelium discoideum JOURNAL Plasmid 61 (2), 110-118 (2009) PUBMED 19063918 REFERENCE 2 (bases 1 to 3518) AUTHORS Veltman DM, Akar G, Bosgraaf L, van Haastert PJM. TITLE Direct Submission JOURNAL Submitted (19-JUL-2008) Cell Biochemistry, Rijks Universiteit Groningen, Kerklaan 30, Haren 9751 NN, The Netherlands REFERENCE 3 (bases 1 to 3518) TITLE Direct Submission REFERENCE 4 (bases 1 to 3518) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; date: "2009"; volume: "61"; issue: "2"; pages: "110-118" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (19-JUL-2008) Cell Biochemistry, Rijks Universiteit Groningen, Kerklaan 30, Haren 9751 NN, The Netherlands" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..3518 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 10..87 /codon_start=1 /label=CBP /note="calmodulin-binding peptide" /translation="KRRWKKNFIAVSAANRFKKISSSGAL" CDS 115..135 /codon_start=1 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" /translation="ENLYFQG" CDS 175..348 /codon_start=1 /label=ProtA /note="IgG-binding unit of Staphylococcus aureus protein A" /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL AEAKKLNDAQAPK" CDS 352..522 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA EAKKLNGAQAPK" primer_bind complement(579..595) /label=SK primer /note="common sequencing primer, one of multiple similar variants" promoter complement(632..650) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(671..687) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(695..711) /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(719..749) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(764..785) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1073..1661) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1835..2692) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2693..2797) /label=AmpR promoter rep_origin 2824..3279 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 3420..3436 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 3446..3464 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 3490..3506 /label=KS primer /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 actagtgaaa agagaagatg gaaaaagaat ttcatagccg tctcagcagc caaccgcttt 61 aagaaaatct catcctccgg ggcacttgat tatgatattc caactactgc tagcgagaat 121 ttgtattttc agggtgagct caaaaccgcg gctcttgcgc aacacgatga agccgtggac 181 aacaaattca acaaagaaca acaaaacgcg ttctatgaga ttttacattt acctaactta 241 aacgaagaac aacgaaacgc cttcatccaa agtttaaaag atgacccaag ccaaagcgct 301 aaccttttag cagaagctaa aaagctaaat gatgctcagg cgccgaaagt agacaacaaa 361 ttcaacaaag aacaacaaaa cgcgttctat gagattttac atttacctaa cttaaacgaa 421 gaacaacgaa acgccttcat ccaaagttta aaagatgacc caagccaaag cgctaacctt 481 ttagcagaag ctaaaaagct aaatggtgct caggcgccga aagtagacgc gaattccgcg 541 gggaagtaat ctagacatcg aattcctgca gcccggggga tccactagtt ctagagcggc 601 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg 661 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 721 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 781 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 841 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 901 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 961 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 1021 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 1081 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 1141 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 1201 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 1261 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 1321 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 1381 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 1441 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 1501 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 1561 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 1621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 1681 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 1741 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 1801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 1861 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 1921 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 1981 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 2041 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 2101 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 2161 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 2221 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 2281 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 2341 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 2401 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 2461 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 2521 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 2581 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 2641 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 2701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 2761 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 2821 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 2881 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 2941 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 3001 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 3061 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 3121 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 3181 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 3241 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg 3301 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 3361 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 3421 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg 3481 gccccccctc gaggtcgacg gtatcgataa gcttgatg //