LOCUS       Exported                6754 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway cloning destination vector pcGFP1Fpac, complete sequence.
ACCESSION   MH048881
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  AUTHORS   Dawson SC, House SA.
  TITLE     Imaging and analysis of the microtubule cytoskeleton in giardia
  JOURNAL   Methods Cell Biol. 97, 307-339 (2010)
  PUBMED    20719278
REFERENCE   2  (bases 1 to 6754)
  AUTHORS   Hagen KD, Hirakawa MP, House SA, Schwartz CL, Pham JK, Cipriano MJ, 
            De La Torre MJ, Sek AC, Du G, Forsythe BM, Dawson SC.
  TITLE     Novel structural components of the ventral disc and lateral crest in
            Giardia intestinalis
  JOURNAL   PLoS Negl Trop Dis 5 (12), e1442 (2011)
  PUBMED    22206034
REFERENCE   3  (bases 1 to 6754)
  AUTHORS   Cipriano MJ, Pham JK, Hagen KD, Dawson SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-MAR-2018) Dept of Microbiology and Molecular Genetics,
            University of California, Davis, One Shields Ave, Davis, CA 95616, 
            USA
REFERENCE   4  (bases 1 to 6754)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6754
                     /organism="Gateway cloning destination vector pcGFP1Fpac"
                     /mol_type="other DNA"
                     /label=derived from Giardia intestinalis
                     /note="derived from Giardia intestinalis"
                     /db_xref="taxon:2480754"
     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"
     misc_feature    1064..2774
                     /label=reading frame A, Gateway
                     /note="reading frame A, Gateway"
     misc_feature    1067..1191
                     /label=attR1
                     /note="attR1"
     protein_bind    1067..1191
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1216..1246
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1300..1959
                     /codon_start=1
                     /gene="cat"
                     /product="chloroamphenicol resistance"
                     /label=cat
                     /protein_id="AYM55693.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            1300..1959
                     /gene="cat"
                     /label=cat
     CDS             2301..2606
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /protein_id="AYM55694.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2647..2771)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2647..2747
                     /label=attR2
                     /note="attR2"
     CDS             2785..3504
                     /codon_start=1
                     /product="EGFP"
                     /label=EGFP
                     /note="fluorescent tag"
                     /protein_id="AYM55695.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     3UTR           3514..3638
                     /gene="alpha tubulin"
     gene            3514..3638
                     /gene="alpha tubulin"
                     /label=alpha tubulin
     5UTR           3717..3919
                     /gene="gamma giardin"
     gene            3717..3919
                     /gene="gamma giardin"
                     /label=gamma giardin
     CDS             3920..4528
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin resistance"
                     /label=pac
                     /protein_id="AYM55696.1"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGALE"
     gene            3920..4528
                     /gene="pac"
                     /label=pac
     CDS             3920..4519
                     /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           4529..4646
                     /gene="glutamate dehydrogenase"
     gene            4529..4646
                     /gene="glutamate dehydrogenase"
                     /label=glutamate dehydrogenase
     promoter        complement(4666..4684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4691..4707)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4848..5303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5635..5739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5740..6600
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin resistance"
                     /label=bla
                     /protein_id="AYM55697.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5740..6600
                     /gene="bla"
                     /label=bla