LOCUS       Exported                4791 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector ASM, complete sequence.
ACCESSION   JQ772530
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  AUTHORS   Jerlstrom-Hultqvist J, Stadelmann B, Birkestedt S, Hellman U, Svard 
            SG.
  TITLE     Plasmid vectors for proteomic analyses in giardia: purification of 
            virulence factors and analysis of the proteasome
  JOURNAL   Eukaryotic Cell 11 (7), 864-873 (2012)
  PUBMED    22611020
REFERENCE   2  (bases 1 to 4791)
  AUTHORS   Jerlstrom-Hultqvist J, Stadelmann B, Birkestedt S, Hellman U, Svard 
            SG.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2012) Department of Cell and Molecular Biology, 
            Uppsala University, BMC, Box 596, Uppsala 751 24, Sweden
REFERENCE   3  (bases 1 to 4791)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4791
                     /organism="Cloning vector ASM"
                     /mol_type="other DNA"
                     /db_xref="taxon:1195239"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             327..989
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AFK93523.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     3UTR           2208..3227
                     /note="glutamate dehydrogenase 3 UTR; derived from Giardia
                     intestinalis"
     CDS             complement(2325..2933)
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=puromycin N-acetyltransferase
                     /protein_id="AFK93524.1"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGALE"
     CDS             complement(2334..2933)
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     5UTR           2934..2981
                     /note="glutamate dehydrogenase 5 UTR; derived from Giardia
                     intestinalis"
     5UTR           2988..3187
                     /note="ornithine carbamoyltransferase 5 UTR; derived from 
                     Giardia intestinalis"
     misc_feature    3200..3313
                     /label=SBP-tag
                     /note="SBP-tag"
     CDS             3200..3313
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     misc_feature    3320..3976
                     /label=glutathione S-transferase
                     /note="glutathione S-transferase"
     CDS             3320..3970
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     misc_feature    3977..4000
                     /label=PreScission protease cleavage site
                     /note="PreScission protease cleavage site"
     CDS             3977..4000
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    4040..4057
                     /label=6xHis-tag
                     /note="6xHis-tag"
     CDS             4040..4057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     3UTR           4058..4133
                     /note="alpha-1 giardin 3 UTR; derived from Giardia 
                     intestinalis"
     promoter        complement(4148..4166)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4176..4192)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4334..4789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"