LOCUS       Exported                8083 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCWP1Fluc, complete sequence.
ACCESSION   MF062157
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8083)
  AUTHORS   Dawson SC.
  TITLE     Giardia colonizes and encysts in high density foci in the murine 
            small intestine
  JOURNAL   mSphere (2017) In press
REFERENCE   2  (bases 1 to 8083)
  AUTHORS   Dawson SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-MAY-2017) Department of Microbiology and Molecular 
            Genetics, Uc Davis, One Shields Avenue, Davis, CA 95616, USA
REFERENCE   3  (bases 1 to 8083)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8083
                     /organism="Cloning vector pCWP1Fluc"
                     /mol_type="other DNA"
                     /label=derived from Giardia intestinalis
                     /note="derived from Giardia intestinalis"
                     /db_xref="taxon:2011122"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    893..914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        929..959
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    967..983
                     /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     991..1007
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1025..1043
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     5UTR           1122..1264
                     /gene="CWP1"
     gene            1122..1264
                     /gene="CWP1"
                     /label=CWP1
     CDS             1271..2923
                     /codon_start=1
                     /product="FLuc"
                     /label=FLuc
                     /note="firefly luciferase"
                     /protein_id="ASA45575.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     3UTR           2934..3403
                     /gene="CWP2"
     gene            2934..3403
                     /gene="CWP2"
                     /label=CWP2
     5UTR           3604..3803
                     /gene="gamma giardin"
     gene            3604..3803
                     /gene="gamma giardin"
                     /label=gamma giardin
     CDS             3807..4415
                     /codon_start=1
                     /product="pac"
                     /label=pac
                     /note="puromycin resistance"
                     /protein_id="ASA45576.1"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGALE"
     CDS             3807..4406
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     3UTR           4416..4534
                     /gene="GDH"
     gene            4416..4534
                     /gene="GDH"
                     /label=GDH
     misc_feature    complement(4539..5255)
                     /label=similar to aurora kinase
                     /note="similar to aurora kinase"
     CDS             complement(5268..6227)
                     /codon_start=1
                     /product="NOP5"
                     /label=NOP5
                     /protein_id="ASA45578.1"
                     /translation="MLAVAPNFTRIVGSIVGVRLLAKAGSLLNLAKLPASTLQILGSER
                     ALFRAKKMRSKQTPKYGFIYHAALVGKASATNRGKMARVVATNASLAARVDALSESSNT
                     SFAKGKLEKINASLRFYEGDKQARSNRKSIASQKMEAYRRGVATGYEGKGDMKLSKETA
                     PAAEPKKTRMTEAFEEDTGFTQTKASQDKGSRNKENAQERQDDAMPAPLEPLSSAAKKD
                     EPVAPIEVDAEDTNDTKDAKEKHKEKSKEKHRDKDKEKEKHKEKEKDKEKHRDKDKDKE
                     RDKEKHKDKDKHRDKDREKDKEKHKSKDDDKKNKTHSK"
     CDS             complement(5987..5998)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(6249..6267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6274..6290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6431..6886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6964..7068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7069..7929
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ASA45577.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"