LOCUS       Exported                5716 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Artificial vector pJL531, complete sequence.
ACCESSION   JF295006
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  AUTHORS   Patel RD, Lodge JK, Baker LG.
  TITLE     Going green in Cryptococcus neoformans: the recycling of a 
            selectable drug marker
  JOURNAL   Fungal Genet. Biol. 47 (3), 191-198 (2010)
  PUBMED    19944774
REFERENCE   2  (bases 1 to 5716)
  AUTHORS   Patel RD, Lodge JK, Baker LG.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-FEB-2011) Molecular Microbiology, Washington 
            University, 660 S Euclid Ave, St. Louis, MO 63110, USA
REFERENCE   3  (bases 1 to 5716)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5716
                     /organism="Artificial vector pJL531"
                     /mol_type="other DNA"
                     /db_xref="taxon:1034055"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     misc_feature    204..225
                     /label=PCR primer binding site 3/4
                     /note="PCR primer binding site 3/4"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    294..353
                     /label=LoxP site
                     /note="LoxP site"
     protein_bind    complement(310..343)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      354..1178
                     /regulatory_class="promoter"
                     /note="Cryptococcus neoformans actin promoter"
     CDS             1179..1754
                     /codon_start=1
                     /product="nourseothricin drug marker protein"
                     /label=nourseothricin drug marker protein
                     /note="NAT"
                     /protein_id="AEF32778.1"
                     /translation="MAAATLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDG
                     FTLREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWN
                     RRLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGF
                     TLCGLDTALYDGTASDGEQALYMSMPCP"
     CDS             1191..1754
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="TLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFTLR
                     EVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRRLT
                     VEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTLCG
                     LDTALYDGTASDGEQALYMSMPCP"
     regulatory      1755..2017
                     /regulatory_class="terminator"
                     /note="Cryptococcus neoformans Trp-1 terminator"
     misc_feature    2018..2079
                     /label=LoxP site
                     /note="LoxP site"
     protein_bind    complement(2038..2071)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(2150..2168)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2173..2194
                     /label=PCR primer binding site 5/6
                     /note="PCR primer binding site 5/6"
     primer_bind     complement(2175..2191)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2332..2760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             3104..3898
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3916..4776
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4947..5535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"