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
//
