LOCUS       Exported                6749 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTREXL-Hyg, complete sequence.
ACCESSION   JN596093
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6749)
  AUTHORS   Bouvier LA, de los Milagros Camara M, Canepa GE, Miranda MR, Pereira
            CA.
  TITLE     Plasmids and molecular building blocks to assist the development and
            extension of genetic manipulation tools for Trypanosoma cruzi
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6749)
  AUTHORS   Bouvier LA, de los Milagros Camara M, Canepa GE, Miranda MR, Pereira
            CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-AUG-2011) Laboratorio de Biologia Molecular de 
            Trypanosoma cruzi, Instituto de Investigaciones Medicas (IDIM) 
            Alfredo Lanari, Combatientes de Malvinas 3150, CABA, Buenos Aires 
            1427, Argentina
REFERENCE   3  (bases 1 to 6749)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6749
                     /organism="Cloning vector pTREXL-Hyg"
                     /mol_type="other DNA"
                     /db_xref="taxon:1133400"
     regulatory      2..811
                     /regulatory_class="promoter"
                     /gene="ribosomal promoter"
     gene            2..811
                     /gene="ribosomal promoter"
                     /label=ribosomal promoter
     intron          862..1033
                     /gene="HX1"
     gene            862..1033
                     /gene="HX1"
                     /label=HX1
     regulatory      1057..1066
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1063..1782
                     /codon_start=1
                     /gene="eGFP"
                     /product="eGFP"
                     /label=eGFP
                     /note="enhanced green fluorescent protein"
                     /protein_id="AEZ51471.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            1063..1782
                     /gene="eGFP"
                     /label=eGFP
     intron          1819..2331
                     /gene="IgapdhI-II"
     gene            1819..2331
                     /gene="IgapdhI-II"
                     /label=IgapdhI-II
     CDS             2332..3357
                     /codon_start=1
                     /gene="Hyg"
                     /product="Hyg"
                     /label=Hyg
                     /note="hygromycin B phosphotransferase"
                     /protein_id="AEZ51472.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     D"
     gene            2332..3357
                     /gene="Hyg"
                     /label=Hyg
     intron          3358..3875
                     /gene="IgapdhII"
     gene            3358..3875
                     /gene="IgapdhII"
                     /label=IgapdhII
     promoter        complement(3912..3930)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3951..3967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3975..3991
                     /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(3999..4029)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4044..4065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4294..4967)
                     /direction=LEFT
     rep_origin      complement(4353..4941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5112..5972)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="beta-lactamase"
                     /protein_id="AEZ51473.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5112..5972)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(5973..6077)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(5973..6071)
                     /regulatory_class="promoter"
                     /gene="bla"
     gene            complement(5973..6071)
                     /gene="bla"
                     /label=bla
     rep_origin      complement(6103..6558)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6700..6716
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6723..6741
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"