LOCUS       40924_14790        3511 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDM229, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3511)
  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 3511)
  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 3511)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3511)
  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..3511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             42..215
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             219..389
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             426..446
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             453..530
                     /codon_start=1
                     /label=CBP
                     /note="calmodulin-binding peptide"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     primer_bind     complement(572..588)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(625..643)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(664..680)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(688..704)
                     /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(712..742)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(757..778)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1066..1654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1828..2685)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2686..2790)
                     /label=AmpR promoter
     rep_origin      2817..3272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3413..3429
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3439..3457
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3483..3499
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggatccaaaa aatggcaggc cttgcgcaac acgatgaagc cgtggacaac aaattcaaca
       61 aagaacaaca aaacgcgttc tatgagattt tacatttacc taacttaaac gaagaacaac
      121 gaaacgcctt catccaaagt ttaaaagatg acccaagcca aagcgctaac cttttagcag
      181 aagctaaaaa gctaaatgat gctcaggcgc cgaaagtaga caacaaattc aacaaagaac
      241 aacaaaacgc gttctatgag attttacatt tacctaactt aaacgaagaa caacgaaacg
      301 ccttcatcca aagtttaaaa gatgacccaa gccaaagcgc taacctttta gcagaagcta
      361 aaaagctaaa tgatgctcag gcgccgaaag tagacgcgaa ttgtgatata cctacaactg
      421 cttctgaaaa tttatatttt caaggtgaac taaagagaag atggaaaaag aatttcatag
      481 ccgtctcagc agccaaccgc tttaagaaaa tctcatcctc cggggcactt gatgatgacg
      541 atagatctga tcgaattcct gcagcccggg ggatccacta gttctagagc ggccgccacc
      601 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
      661 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      721 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      781 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      841 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      901 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1501 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1861 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1921 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1981 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2041 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2101 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2161 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2221 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2281 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2341 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2401 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2461 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2521 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2581 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2641 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2701 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2761 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     2821 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2881 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2941 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3001 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3061 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3121 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3181 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3241 gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt
     3301 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3361 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3421 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     3481 ctcgaggtcg acggtatcga taagcttgat g
//
