LOCUS       Exported                6355 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pTtag, complete sequence.
ACCESSION   FJ789658
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6355)
  AUTHORS   El-Haddad H, Przyborski JM, Kraft LG, McFadden GI, Waller RF, Gould 
            SB.
  TITLE     Characterization of TtALV2, an Essential Charged Repeat Motif 
            Protein of the Tetrahymena thermophila Membrane Skeleton
  JOURNAL   Eukaryotic Cell 12 (6), 932-940 (2013)
  PUBMED    23606287
REFERENCE   2  (bases 1 to 6355)
  AUTHORS   Gould SB, McFadden GI, Waller RF.
  TITLE     Analysis of the two alveolate-characterizing epiplasmin proteins 
            alveolin 1 and 2 from Tetrahymena thermophila
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6355)
  AUTHORS   Gould SB, McFadden GI, Waller RF.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-FEB-2009) School of Botany, University of Melbourne, 
            Professors Walk, Parkville, VIC 3010, Australia
REFERENCE   4  (bases 1 to 6355)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6355
                     /organism="Transformation vector pTtag"
                     /mol_type="other DNA"
                     /db_xref="taxon:629723"
     5UTR           6..520
                     /label=BTU1
                     /note="BTU1"
     regulatory      527..1455
                     /regulatory_class="promoter"
                     /note="MTT promoter"
     misc_feature    1456..1494
                     /label=MCS
                     /note="MCS"
     CDS             1495..2256
                     /codon_start=1
                     /product="GFP"
                     /label=GFP
                     /note="codon optimized for Tetrahymena thermophila"
                     /protein_id="ACO05019.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGV*CFSRYPDHMK*HDFFKSAMPEGYV*ERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADK*KNGIK
                     VNFKIRHNIEDGSV*LADHY**NTPIGDGPVLLPDNHYLST*SALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSRMHHHHHHHHHMH"
     CDS             2221..2247
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     misc_feature    2228..2253
                     /label=HIS-tag
                     /note="HIS-tag"
     3UTR           2257..3490
                     /label=BTU1
                     /note="BTU1"
     misc_feature    3491..6355
                     /label=pPCR backbone
                     /note="pPCR backbone"
     promoter        complement(3509..3527)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3548..3564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3572..3588
                     /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)."
     promoter        complement(3596..3626)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3641..3662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3950..4538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4709..5569)
                     /codon_start=1
                     /transl_except="(pos: 0 .. 0,  aa: TERM)"
                     /product="AmpR"
                     /label=AmpR
                     /note="bacterial beta-lactamase"
                     /protein_id="ACO05020.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5570..5674)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5700..6155)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6297..6313
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6323..6341
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"