LOCUS       Exported                5821 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYL2, complete sequence.
ACCESSION   KU378203
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5821)
  AUTHORS   Pomraning KR.
  TITLE     Yarrowia lipolytica functional genomics toolbox
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5821)
  AUTHORS   Pomraning KR.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JAN-2016) Environmental Molecular Science Lab, Pacific
            Northwest National Lab, 902 Battelle Blvd, Richland, WA 99352, USA
REFERENCE   3  (bases 1 to 5821)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5821
                     /organism="Cloning vector pYL2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1890044"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             343..354
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      382..1382
                     /regulatory_class="promoter"
                     /gene="exp1"
     gene            382..1382
                     /gene="exp1"
                     /label=exp1
     CDS             1431..2144
                     /codon_start=1
                     /product="10xGly-sGFP"
                     /label=10xGly-sGFP
                     /note="10x Glycine linker attached to super-fold green 
                     fluorescent protein"
                     /protein_id="APB62237.1"
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1431..2141
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     regulatory      2175..2579
                     /regulatory_class="promoter"
                     /gene="tef"
     gene            2175..2579
                     /gene="tef"
                     /label=tef
     CDS             2592..3617
                     /codon_start=1
                     /gene="hph"
                     /product="HPH"
                     /label=hph
                     /note="Hygromycin phosphotransferase"
                     /protein_id="APB62236.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     gene            2592..3617
                     /gene="hph"
                     /label=hph
                     /note="codon optimized for Yarrowia lipolytica"
     misc_feature    3662..3906
                     /note="modified version of ARS68, contains centromere and 
                     origin of replication sequences for Yarrowia lipolytica"
     rep_origin      4081..4748
     rep_origin      complement(4137..4725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4899..5693)
                     /codon_start=1
                     /gene="kanR"
                     /product="kanR"
                     /label=kanR
                     /protein_id="APB62235.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(4899..5693)
                     /gene="kanR"
                     /label=kanR
     promoter        complement(5694..5798)
                     /gene="bla"
                     /label=AmpR promoter