LOCUS       Exported                9016 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGDHFluc, complete sequence.
ACCESSION   MF062155
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9016)
  AUTHORS   Barash NR, Nosala C, Pham JK, McInally SG, Gourguechon S, 
            McCarthy-Sinclair B, 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 9016)
  AUTHORS   Barash NR, Nosala C, Pham JK, McInally SG, Gourguechon S, 
            McCarthy-Sinclair B, Dawson SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2017) Department of Microbiology and Molecular 
            Genetics, Uc Davis, One Shields Avenue, Davis, CA 95616, USA
REFERENCE   3  (bases 1 to 9016)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9016
                     /organism="Cloning vector pGDHFluc"
                     /mol_type="other DNA"
                     /label=derived from Giardia intestinalis
                     /note="derived from Giardia intestinalis"
                     /db_xref="taxon:2011124"
     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"
     gene            1050..4171
                     /gene="GDH"
                     /label=GDH
     5UTR           1050..1101
                     /gene="GDH"
     CDS             1104..2756
                     /codon_start=1
                     /gene="GDH"
                     /product="FLUC"
                     /label=GDH
                     /note="firefly luciferase"
                     /protein_id="ASA45568.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2800..2921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     3UTR           3041..3162
                     /gene="alpha-2 tubulin"
     gene            3041..3162
                     /gene="alpha-2 tubulin"
                     /label=alpha-2 tubulin
     5UTR           3241..3440
                     /gene="gamma giardin"
     gene            3241..3440
                     /gene="gamma giardin"
                     /label=gamma giardin
     CDS             3444..4052
                     /codon_start=1
                     /gene="GDH"
                     /product="pac"
                     /label=GDH
                     /note="puromycin resistance"
                     /protein_id="ASA45569.1"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGALE"
     CDS             3444..4043
                     /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           4053..4171
                     /gene="GDH"
     misc_feature    complement(4176..4892)
                     /label=similar to aurora kinase
                     /note="similar to aurora kinase"
     5UTR           complement(4905..6919)
                     /gene="NOP5"
     CDS             complement(4905..6746)
                     /codon_start=1
                     /product="NOP5"
                     /label=NOP5
                     /protein_id="ASA45571.1"
                     /translation="MYLLYEAASGLAMFKVLDESKLKAADLVGAMMRTSSDVQKIVSLV
                     SFSKFPSVEGALATQTSTFEKDVSEELKKFVEKHFVNTVAKKDASEKLLVQDPHMVTAL
                     KDVYGVRAAFNDIVVELSRGIRTHLASILPDFDQEHDSSMAMGLAHSFSRYRVKFNADS
                     IDIMLIQAIALIDDTEKDINQFAMRLREWMVWHFPELITIVPDNIDYARTVVVLRDRRH
                     LEESLTTSGREKFIKALNKASKDQAYAIISAAKTSVGTDITDQDSDRVVHLAEELLSLV
                     SYREELHEYIEARMLAVAPNFTRIVGSIVGVRLLAKAGSLLNLAKLPASTLQILGSERA
                     LFRAKKMRSKQTPKYGFIYHAALVGKASATNRGKMARVVATNASLAARVDALSESSNTS
                     FAKGKLEKINASLRFYEGDKQARSNRKSIASQKMEAYRRGVATGYEGKGDMKLSKETAP
                     AAEPKKTRMTEAFEEDTGFTQTKASQDKGSRNKENAQERQDDAMPAPLEPLSSAAKKDE
                     PVAPIEVDAEDTNDTKDAKEKHKEKSKEKHRDKDKEKEKHKEKEKDKEKHRDKDKDKER
                     DKEKHKDKDKHRDKDREKDKEKHKSKDDDKKNKTHSK"
     CDS             complement(5624..5635)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     gene            complement(6747..6919)
                     /gene="NOP5"
                     /label=NOP5
     promoter        complement(6928..6946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6953..6969)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(7110..7565)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7897..8001
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8002..8862
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance"
                     /protein_id="ASA45570.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"