LOCUS       Exported               14986 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPZ3TA, complete sequence.
ACCESSION   EF667963
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14986)
  AUTHORS   Goodwin TJ, Busby JN, Poulter RT.
  TITLE     A yeast model for target-primed (non-LTR) retrotransposition
  JOURNAL   BMC Genomics 8, 263 (2007)
  PUBMED    17683538
REFERENCE   2  (bases 1 to 14986)
  AUTHORS   Goodwin TJD., Busby JN, Poulter RTM.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUN-2007) Department of Biochemistry, University of 
            Otago, Cumberland Street, Dunedin 9016, New Zealand
REFERENCE   3  (bases 1 to 14986)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14986
                     /organism="Expression vector pPZ3TA"
                     /mol_type="other DNA"
                     /note="Zorro3 target-primed (non-LTR) retrotransposon 
                     expression vector"
                     /db_xref="taxon:463568"
     regulatory      23..1043
                     /regulatory_class="promoter"
                     /note="Candida albicans ACT1 promoter"
     mobile_element  1050..7847
                     /mobile_element_type="retrotransposon:Zorro3"
                     /note="derived from Candida albicans; carries URA3 gene 
                     disrupted by antisense intron"
     repeat_region   1050..1069
                     /label=5 poly-A tract
                     /note="5 poly-A tract"
     CDS             1075..2901
                     /codon_start=1
                     /product="Zorro3 ORF1 protein"
                     /label=Zorro3 ORF1 protein
                     /protein_id="ABU62830.1"
                     /translation="MTTNGTNGFDFADKGDHPPRQSQENPPGNFQELPLDGGQIRKSSS
                     NDGVSNNVEFNTNTNAHESWADYPESDQMDTDSSEYYTEDENEEGKIPKGPYHQFQNAN
                     KFTYFTPETATTLVAPNNNTQNTTTEQDNMSKTYASVAATFKTNLAKSQDKQEELRMSN
                     FKVPQKAFDDLKRHNDAVFPEEVMTSDLSTIINKYGHEKVFDTIENTLITTAQNLEGYL
                     SEYPENVMRKAFKYFTLKLSIEDKQTRDVCEEILGIREELMYIQRILDFNIMDKVMCFM
                     PFTFEQMEGKLSSEKAKVRTIVENLELDFDGTVEHIKIDSAKRNRQLVAFKVPIHCRSL
                     AERYFRQIFLHKEDVIVQQYAPIIEKVQLGRKGKTEENNHQLLTEYVCYFIVDLFSGQV
                     PKRRIKIDNTKNKPSSYEQCTSRVLSNCKYCEYCRSMTHTKYHCPLIKPCTNCGVKGHK
                     TTSCKKALSTNTLVLKRPETKPNFPILPPKVVNKTTSDDSQRKGSIAKTTSHTKGDNTE
                     KMVTPSAQNENTTLQESIPVTITGHREAPTTPPVADTTAKKTMTTGVLEMDTSKHTISS
                     NDEASPRKKLNVKEKPSNPEKDPGLNLPPNLQ"
     CDS             3074..6523
                     /codon_start=1
                     /product="Zorro3 ORF2 protein"
                     /label=Zorro3 ORF2 protein
                     /note="contains putative endonuclease and reverse 
                     transcriptase domains"
                     /protein_id="ABU62831.1"
                     /translation="MKRNESYINSLTIGSKNIGSHQSTDFKKLLDIFLKLIGEHLMIDI
                     WFIQEIRFVSQEQFNYINKILKQHNAQLRMHHFEDLTGFLIHSPHAKLKFKIRDNDNHH
                     TTHFEGRISILDITLITNEDITLINNYLHSGNMDAQMTLLKAFIKYIANLKKHTNHNII
                     YGGDYNHIMLLDDVQLPLDQTRYIISKKELEIIQLMSNFYKKWKLQDAFQIRNNLQPTN
                     FHSNKSVKKRLDRIYIDSRIRRKLRNCRILEEFKQISTHKIIAMSFQIQKEPILKVGNP
                     RYLIPQWMSQDENIIKDLNNNEQSSLTPFSNWNGIINRIKEKVIFYEKYQRYIRAYIPH
                     AGNFPDEKMLKFRFRPSLNFSIITEMQTESGNTVQDTEIMINLATKFYQDLFLVEDRHL
                     ESFTFVEQFDKKIDDTDKVLLEKAFIEENVYDHLLMINKKTAVGTDGISYQNLIELWPS
                     LGESLIRAGNNILKYGTLPQQMSEVIITLIPKKLKTPIIENFRPISVISCAVRLLSSVI
                     EKQLNPVLAKVIEKTQTGFLKERSISNSIYLLDMVLTRYQTSKTADAESAGFINLDFRK
                     AFDTVHHDFILKVLQQVGFGPKATNFLMAITAKQKAKVSINNIEGPCFPLKRGVRQGNP
                     ISPLIFILILETFLARLSKEIEGIGVVNEVSLVAYTAYADDVIIFFKNKNDQERIQQLL
                     EDFGRESGLYLNNNKTEVCYFNDIPEISFLPYVSKKLQLEKLTYLGVPMKKADEEFDPW
                     TLFVLNLNAQIRMTPILDLPYQLIMKLMNIFIFSKLYYRDLHSPILTTAVSSIITTVQQ
                     RLPLYFKLQRLQTPNHLGGFGLMNPNHQVKGRRGKQIYLLYTQEDDLIIKFMRTKIQDI
                     LDNIAKDYYTMPTEPDKLIVYPWYLFGMGVSCHLSLQFRYLKERVYENLTKLEISWFEA
                     WFQLVHYTGPPVETPIIIMLIEDYANLIIQPQTESIKLMSPNFEEQSLSEQTFHHTSRK
                     LCPQAPIIPEGWGKHFDTIKSLTISDWTEFWKNMNKVQKQSVGSLQDYHLFILGYYSHY
                     PFYPNYKKLEIHFCQLCNTGTDSIVHHIFECGETMELWLKHFNSQRTPQFIIGNKHLHK
                     KDLYALNEYIKEVVQKVKRRRGSPILNQGERENVDAGTNVLV"
     3UTR           join(6524..6630,7684..7847)
                     /label=Zorro3 3 UTR
                     /note="Zorro3 3 UTR"
     CDS             complement(join(6646..7449,7534..7542))
                     /codon_start=1
                     /gene="URA3"
                     /allele="URA3i"
                     /product="orotidine-5-monophosphate decarboxylase"
                     /label=URA3
                     /note="derived from Candida albicans"
                     /protein_id="ABU62832.1"
                     /translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
                     FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
                     KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
                     AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
                     TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRDAGWNAYLKKTGQL"
     gene            complement(6646..7542)
                     /gene="URA3"
                     /allele="URA3i"
                     /label=URA3
     intron          7450..7533
                     /note="intron from Candida albicans peptide transporter in 
                     GenBank Accession Number U09781"
     regulatory      complement(7543..7677)
                     /regulatory_class="promoter"
                     /note="Candida albicans URA3 promoter"
     repeat_region   7828..7847
                     /label=3 poly-A tract
                     /note="3 poly-A tract"
     misc_feature    7848..7998
                     /label=Zorro3 3 flanking sequence
                     /note="Zorro3 3 flanking sequence"
     misc_feature    8004..8394
                     /note="ACT1 terminator; derived from Candida albicans"
     gene            complement(8401..11095)
                     /gene="IMH3"
                     /label=IMH3
                     /note="derived from Candida albicans"
     CDS             complement(join(8685..9799,10047..10497))
                     /codon_start=1
                     /gene="IMH3"
                     /product="mycophenolic acid resistance protein"
                     /label=IMH3
                     /protein_id="ABU62833.1"
                     /translation="MVFETSKATSYLKDYPKKDGLSVKELIDSTNFGGLTYNDFLILPG
                     LINFPSSAVSLETKLTKKITLKSPFVSSPMDTVTEENMAIHMALLGGIGIIHHNCTSEE
                     QAEMVRKVKKYENGFINDPVVISPEVTVGEVKKMGEVLGFTSFPVTENGKVGGKLVGII
                     TSRDIQFHEDNKSPVSEVMTKDLVVGKKGISLTDGNELLRSSKKGKLPIVDAEGNLVSL
                     ISRTDLQKNQDYPNASKSFHSKQLLCGATIGTIDADRERLDKLVEAGLDVVVLDSSNGS
                     SVFQLNMIKWIKEKYPELQVIAGNVVTREQAALLIEAGADALRIGMGSGSICITQEVMA
                     CGRPQGTAVYGVTEFANKFGVPCIADGGIGNIGHITKALALGASCVMMGGLLAGTAETP
                     GDYFYRDGKRLKTYRGMGSIDAMQQTNTNANASTSRYFSEADKVLVAQGVSGSVVDKGS
                     ITKFVPYLYNGLQHSLQDIGIKSIDELRENVDNGEIRFEFRTASAQFEGGVHGLHSYEK
                     RLHN"
     gene            11102..12340
                     /gene="RP10"
                     /label=RP10
                     /note="derived from Candida albicans"
     CDS             11385..12155
                     /codon_start=1
                     /gene="RP10"
                     /product="RP10 protein"
                     /label=RP10
                     /protein_id="ABU62834.1"
                     /translation="MAVGKNKRLSKGKKGLKKKVVDPFTRKDWLDIKAPTTFENRNVGK
                     TLINRSTGLKNAADGLKGRVFEVCLADLQGSEDHSYRKIKLRVDEVQGKNLLTNFHGLD
                     FTSDKLRSLVRKWQSLVEANVTVKTSDDYVLRVFAIAFTKRQPNQIKKTTYAQSSKLRE
                     VRKKMIEIMQREVSNCTLAQLTSKLIPEVIGREIEKSTQTIFPLQNVHIRKVKLLKQPK
                     FDLGSLLALHGEGSTEEKGKKVSSGFKDVVLESV"
     misc_feature    12342..14986
                     /label=derived from pUC19 vector
                     /note="derived from pUC19 vector"
     primer_bind     complement(12370..12386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    12394..12410
                     /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(12418..12448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12463..12484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(12772..13360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(13531..14391)
                     /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"
     promoter        complement(14392..14496)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     14970..14986
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"