LOCUS       Exported                5544 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPOP, complete sequence.
ACCESSION   AF323603
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5544)
  AUTHORS   Soderholm JF, Bevis BJ, Glick BS.
  TITLE     A vector for Pop-in/Pop-out gene replacement in Pichia pastoris
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5544)
  AUTHORS   Soderholm JF, Bevis BJ, Glick BS.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-NOV-2000) Molecular Genetics and Cell Biology, 
            University of Chicago, 920 E. 58th St., Chicago, IL 60637, USA
REFERENCE   3  (bases 1 to 5544)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5544
                     /organism="Cloning vector pPOP"
                     /lab_host="Pichia pastoris"
                     /mol_type="genomic DNA"
                     /note="vector used for pop-in pop-out gene replacement in 
                     Pichia pastoris"
                     /db_xref="taxon:154146"
     CDS             complement(302..886)
                     /codon_start=1
                     /product="T-urf13"
                     /label=T-urf13
                     /protein_id="AAK19282.1"
                     /translation="MASTRVLASRLASQMAASAKVARPAVRVAQVSKRTIQTGSPLQTL
                     KRTQMTSIVNATTRQAFQKRAYSSEIAQAMARSGMITTFLNLPPFDQGLVFFGSIFIFF
                     LCILLIKGYLRKMDDSYLAQLSELANHNRVEAAKAGHVALHELSFSWLRGVQIRVRTLP
                     IQWNEGGGRSNDQSTLSKPKYSSMTDSVQVP"
     promoter        complement(900..1376)
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             complement(1280..1675)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRILESILFCRN
                     VLVSSSNQVAISEISGSVATPNDLLAT"
     primer_bind     1585..1601
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1602..1660
                     /label=pUC19 multiple cloning site
                     /note="pUC19 multiple cloning site"
     misc_feature    1602..1658
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1671..1687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1695..1711
                     /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(1719..1749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1764..1785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2073..2661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2832..3692)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AAK19284.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        3710..4025
                     /gene="S. cerevisiae ARG4"
                     /label=ARG4 promoter
     CDS             4026..5417
                     /codon_start=1
                     /product="Arg4"
                     /label=Arg4
                     /protein_id="AAK19283.1"
                     /translation="MSDGTQKLWGGRFTGETDPLMHLYNASLPYDYKMYKADLEGTKVY
                     TAGLQKLGLLTETELAKIHEGLAEIKKEWDADKFVRHPNDEDIHTANERRLGELIGREI
                     AGKVHTGRSRNDQVVTDLRIYCRDIVNDTLFPALKGLVEVLIKRAEGEIDVLMPGYTHL
                     QRAQPIRWSHWLSSYATYFTEDYKRLGQILHRLNQSPLGAGALAGHPYGIDREFLAEGL
                     GFNSVIGNSLVAVSDRDFIVELMFWGTLFMNHISRFAEDLIIYCTAEFGFIQLSDAYST
                     GSSLMPQKKNADSLELLRGKSGRVFGDLTGFLMSLKGIPSTYDKDMQEDKEPLFDCLTT
                     VEHSMLIATGVISTLTVNKEKMEAALTMDMLATDLADYLVRKGVPFRETHHISGECVAT
                     AERLGLSGIDKLTLEQYQKIDSRFGQDLFETFNFEQSVERRDATGGTAKSAVLKQLDNL
                     KSQLN"