LOCUS       Exported               10779 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Transformation vector pBHRF-108-Ypet, complete sequence.
ACCESSION   MG881769
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10779)
  AUTHORS   Moody LA, Kelly S, Coudert Y, Nimchuk ZL, Harrison CJ, Langdale JA.
  TITLE     Somatic hybridization provides segregating populations for the 
            identification of causative mutations in sterile mutants of the moss
            Physcomitrella patens
  JOURNAL   New Phytol. (2018) In press
  PUBMED    29498048
REFERENCE   2  (bases 1 to 10779)
  AUTHORS   Coudert Y, Nimchuk ZL, Harrison CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JAN-2018) Department of Plant Sciences, University of 
            Oxford, South Parks Road, Oxford OX1 3RB, UK
REFERENCE   3  (bases 1 to 10779)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10779
                     /organism="Transformation vector pBHRF-108-Ypet"
                     /mol_type="other DNA"
                     /db_xref="taxon:2126435"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    182..1293
                     /note="108 targeting sequence for ectopic integration of 
                     transgenes"
     misc_recomb     1363..1396
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(1363..1396)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      1415..1870
                     /regulatory_class="promoter"
                     /note="35S"
     promoter        1599..1944
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1977..3023
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=hygromycin B phosphotransferase
                     /note="confers resistance to hygromycin"
                     /protein_id="AVO64727.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPGSS
                     RVDLQACR"
     regulatory      3024..3198
                     /regulatory_class="terminator"
                     /note="35S terminator"
     regulatory      complement(3232..4013)
                     /regulatory_class="terminator"
                     /note="OCS terminator"
     promoter        3242..3260
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     terminator      3293..3999
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(4015..4737)
                     /codon_start=1
                     /product="YPet"
                     /label=YPet
                     /protein_id="AVO64728.1"
                     /translation="MAAAKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGI
                     KANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLFKDPNEKRDHMVL
                     LEFLTAAGITEGMNELYK"
     misc_feature    complement(4738..5081)
                     /label=29-1
                     /note="29-1"
     regulatory      complement(5098..7119)
                     /regulatory_class="promoter"
                     /note="ZmUbi-1"
     protein_bind    7086..7110
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    7120..8120
                     /note="108 targeting locus for ectopic integration of 
                     transgenes"
     promoter        complement(8138..8156)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8171..8187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8195..8211
                     /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(8219..8249)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8264..8285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8573..9161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9332..10192)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to beta-lactam antibiotics"
                     /protein_id="AVO64729.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(10193..10297)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      10324..10779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"